INSTRUMENT MANUAL

1450-932-10
June 2004

MicroBeta TriLux

and

MicroBeta JET

Liquid Scintillation and Luminescence Counters

Wallac 1450

MicroBeta TriLux

and

MicroBeta JET

Liquid Scintillation and Luminescence

Counters
Valid for MicroBeta TriLux with software version 4.7 and MicroBeta
JET with software version 5.2

PerkinElmer Life and Analytical Sciences, Wallac Oy, P.O. Box 10, FIN-20101 Turku, Finland.
Tel: 358-2-2678111. Fax: 358-2-2678 357. Website: www.perkinelmer.com

Warning
This equipment must be installed and
used in accordance with the
manufacturer's recommendations.
Installation and service must be
performed by personnel properly
trained and authorized by PerkinElmer
Life and Analytical Sciences.
Failure to follow these instructions may
invalidate your warranty and/or impair
the safe functioning of your equipment.

Contents

Contents
Part 1 Introduction
1. Introduction .............................................................................................................................. 7
Operating MicroBeta TriLux - short instructions ....................................................................... 9
Operating MicroBeta JET - short instructions .......................................................................... 11
Part 2 Operating information
2.1 Beginning operation of MicroBeta TriLux .......................................................................... 15
2.2 Cassettes ............................................................................................................................... 25
2.3 Clock setting......................................................................................................................... 27
2.4 Coding cassettes and filtermats ............................................................................................ 29
2.5 Counting ............................................................................................................................... 33
2.6 Counting control................................................................................................................... 37
2.7 Crosstalk correction ............................................................................................................. 41
2.8 Datafiles ............................................................................................................................... 49
2.9 Detectors .............................................................................................................................. 53
2.10 Diskettes ............................................................................................................................. 57
2.11 DPM counting .................................................................................................................... 61
2.12 DPM Standardization ......................................................................................................... 63
2.13 Errors.................................................................................................................................. 69
2.14 Half-life .............................................................................................................................. 71
2.15 Help and Info...................................................................................................................... 73
2.16 Interrupt.............................................................................................................................. 75
2.17 Loading the cassette rack ................................................................................................... 77
2.18 Luminescence counting...................................................................................................... 79
2.19 Microtitration plate format output...................................................................................... 83
2.20 Micro-volume LSC ............................................................................................................ 87
2.21 Monitors and flags.............................................................................................................. 93
2.22 MultiCalc operation ........................................................................................................... 95
2.23 Normalization................................................................................................................... 103
2.24 P-32 Dot blot quantification............................................................................................. 109
2.25 Protocols........................................................................................................................... 111
2.26 Results .............................................................................................................................. 127
2.27 Robotic loading interface ................................................................................................. 139
2.28 Safety and radioactive materials ...................................................................................... 141
2.29 Statistics ........................................................................................................................... 143
2.30 System .............................................................................................................................. 147
2.31 Terminal emulators .......................................................................................................... 159
2.32 Thermostat option ............................................................................................................ 175
2.33 Total count rate ................................................................................................................ 177

Contents

2.34 Window settings............................................................................................................... 179

Part 2J Operating information for MicroBeta JET
2J.0 Injector setup .................................................................................................................... 183
2J.1 Beginning operation of MicroBeta JET............................................................................ 187
2J.6 Counting control ............................................................................................................... 197
2J.18 Luminescence counting .................................................................................................. 201
2J.23 Normalization ................................................................................................................. 207
2J.25 Protocols ......................................................................................................................... 215
2J.27 Robotic loading interface................................................................................................ 231
2J.30 System ............................................................................................................................ 233
Part 3 Instrument description
3.1 Instrument description........................................................................................................ 249
3.2 Routine maintenance .......................................................................................................... 253
3.3 Calculation methods........................................................................................................... 255
3.4 Specifications ..................................................................................................................... 265
3.5 Abbreviations and acronyms used...................................................................................... 277
Declaration of conformity for CE-marking.............................................................................. 283
Part 4 Installation information
4.1 Installation instructions ...................................................................................................... 287
4.2 Installation of the Injector System to MicroBeta JET........................................................ 301
Part 5 Index
5 Index...................................................................................................................................... 309

Trademarks

Trademarks
MicroBeta and MultiCalc are registered trademarks and Betaplate, RiaCalc, OptiPhase and
Wallac are trademarks of PerkinElmer, Inc.
IBM, IBM PC AT, IBM PC XT and PS/2 CGA, EGA, MCGA, VGA and PC-DOS are
trademarks or registered trademarks of International Business Machines Corporation
MS-DOS and Microsoft are registered trademarks of Microsoft Corporation
Amersham is a trademark of Amersham International plc
Olivetti is a trademark of Ing. C Olivetti & C., S.p.A.
Epson is a registered trademark of Epson Corporation
VT is a trademark of Digital Equipment Corporation
Intel is a registered trademark of Intel Corporation
Macintosh is a registered trademark and System 7 and MultiFinder are trademarks of Apple
Computer, Inc.
MultiScreen is a trademark of Millipore Corporation

Part 1 Introduction

1. Introduction

1. Introduction
1.1 Guide to the Instrument manual
There are two versions of Wallac 1450 MicroBeta from PerkinElmer Life and Analytical
Sciences described in this manual: MicroBeta TriLux and MicroBeta JET. Most of the
information is common to both models, where there is a difference it is noted. In Part 2 of this
manual, alternative chapters with the number 2J.xx are included to describe JET features. The
xx is the same number as the equivalent chapter for MicroBeta TriLux.
Normal start-up is described in section 1.2. On the following pages you will find compressed
operating guides for MicroBeta TriLux and MicroBeta JET. Operations include references to
the appropriate chapter in part 2 or part 2J of this manual where you will find a more detailed
explanation. If you are using MicroBeta workstation software, refer to the separate User
manual instead of section 2 in this manual. You only need to refer to section 2 if you work in
terminal mode.
Part 3 of the manual describes the instrument, including specifications. There is also chapter
describing the routine maintenance to be done by the user.
Part 4 is only needed during installation.
Part 5 is the alphabetical index.

1. Introduction

1.2 Instrument start-up

1. Switch on the printer (1a or 1b depending on your configuration), see the appropriate figure
above.
2. Switch on the PC and start up the terminal emulator program or start up MultiCalc and go to
the terminal option by pressing F1 (=COUNTER) followed by F3 (=TERMINAL). See
chapters 2.31 and 2.22 respectively. If you are using the workstation, start the program running
as described in the separate User manual.
3. Insert the MicroBeta program disk into disk drive A, the lower disk drive of the counter, and
the protocol disk into disk drive B, the upper disk drive.
4. Switch on MicroBeta with the power switch at the back of the counter. Loading takes some
3 minutes, then the counter is ready for operation.

Operating MicroBeta TriLux

(See chapters in italics for more information on any subject)
Position
A1

Cassette 1450-101

Function code
Cassette number
Count, norm. or stand. Prot. No.
MultiCalc Assay protocol No.

1 Fix code labels to sample cassettes (2.4)

2 Load cassettes into MicroBeta. Close the
cassette rack compartment door (2.17)
3 To start counting (2.5):
Use the Windows Workstation
Alternatively press:
For MultiCalc: F1 (COUNTER) then Enter
For terminal etc: Protocol number or A
For interrupt or power failure info. see (2.16) and (2.13)

4 The results (2.21, 2.26, 2.32) are sent to

selected output devices (2.8, 2.29)
Terminal or PC (86 486 or Pentium)
Types: VT52/VT100
GenTerm
UltroTerm
MultiCalc
Workstation

Results are sent

to MicroBeta
drive B during
terminal
operation. For
diskette
information see
(2.10)

The printer
connected to
MicroBeta port
3 is used for
terminal and
terminal
emulation
printing.

For System setting info. e.g.

printer or terminal type (2.30)
For Help type H, h or ? (2.15)

Output
to
external
computer e.g a
mainframe.
Connection
to port
2 or PC.

Results go to PC
connected to port
1 in terminal
emulation &
MultiCalc. See
(2.10) for diskette
info. MultiCalc
printout goes to
the printer
connected to the
PC.

Local
area
network
connection
(LAN)
using
the PC.

Code labels (barcodes) are fixed

to an ID-support plate which is
clipped onto a cassette to show
MicroBeta the counting protocol
to use. (2.4)
A counting protocol is a set of
parameters e.g time, isotope,
single/ dual label etc. which
control counting. (2.24, 2.6, 2.7
2.12, 2.14, 2.19, 2.33 )
Counting protocols use either
CPM normalization (2.23) or
DPM standardization (2.12)
info. without or with crosstalk
correction (2.7). Normalization
ensures the counting efficiency
of each detector is the same.
Standardization additionally
calculates the amount of
chemical quenching. Possible
crosstalk between samples can
be corrected for. Normalization
and standardization each require
counting a special set of samples.
This will normally be done
separately from routine
operation. The results will be
stored in MicroBeta and called
into use in normal counting by
means of the counting protocol.
You can use the workstation, a
terminal, a terminal emulator or
MultiCalc to control MicroBeta.
The workstation is described in
a separate user manual.
MultiCalc (2.22, 2.31) can be
used to make assay protocols. An
assay protocol tells which
counting protocol MicroBeta is
to use and what should be done
with the data when it is obtained
e.g. quality control analysis.
Terminal mode is available with
MultiCalc and should be used if
you want to edit normalizations,
standardizations and counting
protocols from within MultiCalc.
With a terminal (2.31) you can
get results to your screen, print
them out or store them on a disk
in MicroBeta.
A terminal emulator (2.31) is a
program which makes a PC act
as a terminal. You can have
results stored on your PC. You
can also use your PC for other
tasks during counting.
Entering commands into
MicroBeta is described in (2.1)
MultiCalc has its own manual.

10

Operating MicroBeta JET

(See chapters in italics for more information on any subject)
Position
A1

Cassette 1450-101

Function code
Cassette number
Count, norm. or stand. Prot. No.
MultiCalc Assay protocol No.

1 Set up injectors (2J.0)

2 Fix code labels to sample cassettes (2.4)
3 Load cassettes into MicroBeta. Close the
cassette rack compartment door (2.17)
4 To start counting (2.5):
Use the Windows Workstation
Alternatively press:
For MultiCalc: F1 (COUNTER) then Enter
For terminal etc: Protocol number or A
For interrupt or power failure info. see (2.16) and (2.13)

5 The results (2.21, 2.26, 2.32) are sent to

selected output devices (2.8, 2.29)
Terminal or PC (86 486 or Pentium)
Types: VT52/VT100
GenTerm
UltroTerm
MultiCalc
Workstation

Injector assembly
fits here with the
module(s) below.
Port 3 is used for
connection to the
injector modules.
Settings: Baud 6,
Parity 1, Data bits
8, Stop bits 2,
Handshake 1, see
(2J.30)

Results are sent

to MicroBeta
drive B during
terminal operation. For diskette
information see
(2.10)

Output
to ext.
computer e.g a
mainframe.
Connect
to port
2 or PC.

Local
area
network
connection
(LAN)
using
PC.

For System setting info. e.g. printer or

terminal type (2.30)
For Help type H, h or ? (2.15)

11

Results go to
PC connected
to port 1 in
terminal
emulation &
MultiCalc. See
(2.10) for
diskette info.
MultiCalc
printout goes to
printer connected to the PC.

Code labels (barcodes) are fixed

to an ID-support plate which is
clipped onto a cassette to show
MicroBeta the counting protocol
to use. (2.4)
A counting protocol is a set of
parameters e.g time, isotope,
single/ dual label etc. which
control counting. (2.24, 2.6, 2.7
2.12, 2.14, 2.19, 2.33 )
Counting protocols use either
CPM normalization (2.23) or
DPM standardization (2.12)
info. without or with crosstalk
correction (2.7.) Normalization
ensures the counting efficiency
of each detector is the same.
Standardization additionally
calculates the amount of
chemical quenching. Possible
crosstalk between samples can
be corrected for. Normalization
and standardization each require
counting a special set of samples.
This will normally be done
separately from routine
operation. The results will be
stored in MicroBeta and called
into use in normal counting by
means of the counting protocol.
You can use the workstation, a
terminal, a terminal emulator or
MultiCalc to control MicroBeta.
The workstation is described in
a separate user manual.
MultiCalc (2.22, 2.31) can be
used to make assay protocols. An
assay protocol tells which
counting protocol MicroBeta is
to use and what should be done
with the data when it is obtained
e.g. quality control analysis.
Terminal mode is available with
MultiCalc and should be used if
you want to edit normalizations,
standardizations and counting
protocols from within MultiCalc.
With a terminal (2.31) you can
get results to your screen, print
them out or store them on a disk
in MicroBeta.
A terminal emulator (2.31) is a
program which makes a PC act
as a terminal. You can have
results stored on your PC. You
can also use your PC for other
tasks during counting.
Entering commands into
MicroBeta is described in (2.1)
MultiCalc has its own manual.

12

Part 2 Operating information

This part has information common to both MicroBeta TriLux and MicroBeta JET and also
information only valid for MicroBeta TriLux. In the latter case there are parallel chapters at the
end of this part which are valid only for MicroBeta JET. These chapters are given the same
number as the MicroBeta TriLux chapter but with a J after the 2 e.g. 2J.1 is the chapter giving
MicroBeta JET information corresponding to the MicroBeta TriLux information in chapter 2.1.
There is information at the head of each MicroBeta TriLux chapter referring you to the relevant
MicroBeta JET chapter.

13

14

2.1 Beginning operation of MicroBeta TriLux

2.1 Beginning operation of MicroBeta TriLux

(See chapter 2J.1 for JET information)
The following conventions are followed all through the MicroBeta user interface:

2.1.1 Menu selections

Menu selections can be entered when the system is in one of the following states:
Ready>
Protocols>
Count>
Info>
System>
Counting protocol>
CPM normaliz. protocol>
DPM standard. protocol>
Assay protocol>
Conveyor>
Total count rate>

=
=
=
=
=
=
=
=
=
=
=

Ready for next operation

Protocol operations
Counting operations
Additional info.
System level operations
Counting protocol editor
CPM norm. protocol editor
DPM std. Protocol editor
MultiCalc assay protocol operation
Conveyor operation
Total count rate manipulation

Command options available, e.g. those shown below, are displayed before the prompt text, e.g.
Ready:
(H)elp
(C)ount

(I)nfo
(P)rotocols

(S)ystem

Ready>

The command is executed by pressing the letter (either lower or upper case) enclosed inside
parentheses. However, commands that include a protocol number (e.g. giving a pure protocol
number in the Ready state to start counting of that protocol) must be completed by pressing the
Enter key.
Help text can be obtained by pressing H, h or ?. Exit to a higher level by pressing Q. Pressing
Control-E (the Ctrl and the E keys at the same time) exits always to the Ready state.

2.1.2 Confirming questions and messages

Quite often MicroBeta displays a message such as:
Press any key when ready or / to exit ->

15

2.1 Beginning operation of MicroBeta TriLux

This kind of message is used to give you time to think if this is really the correct operation and
to check that everything is prepared for the operation. Press any key (such as the space bar)
when ready to continue. Otherwise you can cancel the operation by pressing slash /.
Messages such as:
Do you accept the new terminal setting? (Y/N) ->

are used to notify you that the operation under question can affect the functioning of the
instrument, counting results etc. Press Y=Yes to accept the operation or N=No to cancel it.

2.1.3 Input of data

The rest of the user interface consists mostly of data input. Data input differs somewhat from
menu selection and confirming questions.
- The default or the current value is usually displayed before an arrow sign ->.
- After typing the new value you must press Enter.
- Press the Enter key alone to leave the default or current value unchanged.
- To get help about the subject under question press Enter after H, h or ?.
- Press slash "/" or backslash "\" to exit input mode. In the protocol editor "/" exits with saving
the protocol and "\" without saving it. Elsewhere usually both cause exit without saving the
data.
- Input data can be edited (see below).

2.1.4 Editing input data

If you make a mistake when entering a new input value, you can use the Backspace key
(sometimes labelled as BS or a thick left arrow) or the DEL-key to erase the previous
character. If the input data is quite long this can be tedious. For this kind of situation a simple
keyboard editing facility is provided.
Keyboard editing only uses a few keys so it is easy to learn. The default value or the current
value of input data is used as a template for entering a new value. So if you notice that you
have made a mistake during an early phase of entering a long input (such as output selection in

16

2.1 Beginning operation of MicroBeta TriLux

the protocol editor) you can press Enter to complete entering a new value and then go back to
the same line to correct the mistake.
The editing keys are:
<- or Control-B Go back one character in the template (default or current value).
-> or Control-N Go to the next character in the template.
E.g. Control-B is entered by first pressing down the Control key (usually labelled with Ctrl)
and then typing letter B (lower case or upper case does not matter). If you keep pressing both
the Ctrl-key and letter B down for a while then the repeat function of the terminal sends several
Control-Bs to MicroBeta.
Note that Control-B and Control-N move only inside the default or the current value (displayed
before ->), not inside a newly typed text.
The cursor control keys (left and right arrows) can sometimes be used instead of Control-B and
Control-N. This does not work with every terminal emulator. The up and down arrow keys can
be used when moving up and down in the protocol editor or you can type Lx to jump to line
number x. Please note that the Enter key must be pressed if making changes to any row.
As an example of keyboard editing, suppose you have entered the following printout selection
(the second line):
Printer output: POS CTIME CCPM1 CCPM1%
-> POS CTIMR COUNTS1 CCPM1 CCPM1%

After you press Enter you will see the text:

Error in output heading : CTIMR

and an arrow -> will appear. Press the Control-N keys. As you do this, you will see the letters
POS CTIMR COUNTS1 etc. appearing. After CCPM1% has been displayed the program will
beep to inform that the end of the current value has been reached. Keep Control-B pressed
down and you will see that characters at the end of output selection start disappearing.
Continue until the cursor blinks just after CTIMR (_ shows the place of the cursor):
-> POS CTIMR_

17

2.1 Beginning operation of MicroBeta TriLux

If you go too far then stop pressing Control-B and press Control-N to make the lost characters
visible again. When the cursor is placed just after CTIMR (as in the example) press Backspace
or the DEL key once and R will disappear. Type the letter E.
-> POS CTIM_

(Backspace or DEL pressed)

-> POS CTIME_

(Letter E typed)

Keep pressing Control-N until there is a beep and you get:

-> POS CTIME COUNTS1 CCPM1 CCPM1%

You can now continue typing new headings after CCPM1%. E.g.:
-> POS CTIME COUNTS1 CCPM1 CCPM1% MEAN1 CV1%

Now press Enter to complete editing.

If you later want to remove the heading COUNTS1, go to line 31 to get the programmable
output selection. Keep pressing Control-N until you are just past COUNTS1:
-> POS CTIME COUNTS1_

Press Backspace or DEL 8 times so that COUNTS1 and the preceding space disappears:
-> POS CTIME_

Keep pressing Control-N until there is a beep and press Enter:

-> POS CTIME CCPM1 CCPM1% MEAN1 CV1%_

You can also add a new heading afterwards. Go to programmable output selection again. Keep
pressing Control-N until the cursor is just after CTIME:
-> POS CTIME_

Now type space and the text SQP(I):

-> POS CTIME SQP(I)_

Keep pressing Control-N until the program beeps and then press Enter:
-> POS CTIME SQP(I) CCPM1 CCPM1% MEAN1 CV1%_

18

2.1 Beginning operation of MicroBeta TriLux

Note: Keyboard editing works with most of the type (2.1.3) data input

2.1.5 System operation

The figure following is the flow diagram of the whole MicroBeta operating system. It shows
how the control letters inside parentheses lead to various functions. The functions are described
briefly on the following pages.

19

2.1 Beginning operation of MicroBeta TriLux

Flow chart of MicroBeta TriLux commands (normal level)

Info >
(H)elp
(Q)uit
(U)sage
(C)onsumables
(P)ositions
Ch(a)nges
Cu(s)tomizing
S(y)stem

Ready >
(H)elp
(I)nfo
(C)ount
(P)rotocols
(S)ystem

Count >
(H)elp
(Q)uit
(nn) (count.prot.no)
(?)
(Nnn) (norm.prot.no)
(N?)
(Dnn) (std.prot.no)
(D?)
(A)utomatic counting
Operate con(v)eyor
(T)otal count rate

Protocols >
(H)elp
(Q)uit
(C)ounting protocol
CPM (n)ormalization prot.
DPM (s)tandardization prot.
(M)ultiCalc assay protocol

System >
(H)elp
(Q)uit
MS-D(O)S
(E)rror beep
(S)tatus display
(V)ersion
(C)lock
Se(t)up mode
(R)S-232C
(P)rinter
(D)ata drive
Ter(m)inal
P(a)sswords
Customi(z)ing
(-)Detector temp.

Conveyor>
(H)elp
(Q)uit
(O)ff
(C)lear conveyor
Rack (u)p
Rack (d)own
Total count rate>
(H)elp
(Q)uit
(S)how
(P)rint
(R)eset
(D)efine efficiencies
Counting protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
CPM norm. protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
(N)ormalization data
DPM stand. protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
(R)eplot
MultiCalc assay protocol >
(H)elp
(Q)uit
(S)how
(P)rint
(L)ist
MicroBeta A:\ >
MB - Back to Ready >

20

2.1 Beginning operation of MicroBeta TriLux

MicroBeta TriLux commands (normal level) with a brief description of each

Operations in the Ready State
(H)elp
- Display Help text
(I)nfo
- Get information on using the instrument
(C)ount
- Start counting or operate conveyor
(P)rotocols
- Edit counting parameters
(S)ystem
- Edit system parameters
Count state operations are available in the Ready state too.
Operations in the Info State
(H)elp
(Q)uit
(U)sage
(C)onsumables
(P)ositions
Ch(a)nges
Cu(s)tomizing
S(y)stem

- Display Help text

- Back to the Ready state
- View a short user's manual
- View a list of consumables available
- View normalization and standardization positions
- View a list of changes after the previous version
- View information on how to customize the program
- View information about the system

Operations in the Count State

(H)elp
(Q)uit
(nn) (count.prot.no)
(?)
(Nnn) (norm.prot.no)
(N?)
(Dnn) (std.prot.no)
(D?)
(A)utomatic counting
Operate con(v)eyor
(T)otal count rate
(M)anual counting

- Display Help text

- Back to the Ready state
- Start automatic counting by giving protocol number
- Start automatic counting by selecting protocol from list
- Start CPM normalization by giving protocol number
- Start CPM normalization by selecting protocol from list
- Start DPM standardization by giving protocol number
- Start DPM standardization by selecting protocol from list
- Start automatic counting using IDs
- Conveyor operations
- Show and reset total counts
- Start manual counting (only at Test level)

Operations in the Protocols State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(C)ounting protocol
- Edit counting protocol
CPM (n)ormalization prot.
- Edit CPM normalization protocol
DPM (s)tandardization prot.
- Edit DPM standardization protocol
(M)ultiCalc assay protocol
- Show and print MultiCalc assay protocol

21

2.1 Beginning operation of MicroBeta TriLux

Operations in the Counting Protocol State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(E)dit
- Edit or create and edit a protocol
(nn) (= prot.no)
- Edit a protocol by giving protocol number
(D)elete
- Delete a protocol
(S)how
- Show protocol parameters
(P)rint
- Print a protocol
(L)ist
- Print list of protocols
De(f)ault
- Give default values to a protocol
(C)opy
- Copy one protocol to another
Operations in the CPM Normalization Protocol State
The same operations as in the Counting protocol state, plus:
(N)ormalization data
- Print normalization data
Operations in the DPM Standardization Protocol State
The same operations as in the Counting protocol state, plus:
(R)eplot
- Plot DPM standardization curve
Operations in the MultiCalc Assay Protocol State
Contains only operations (S)how, (P)rint and (L)ist.
Operations in the Conveyor State
(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(O)ff
- Stop conveyor
(C)lear conveyor
- Move cassette to rack and move rack to the middle
Rack (u)p
- Move the rack up one level
Rack (d)own
- Move the rack down one level
Test level contains also:
Remove ca(s)s
(N)ext cass
(P)rev cass
Cu(r)rent cass
Ne(x)t pos
(G)oto pos
C(l)ose detector
Op(e)n detector
Next le(v)el

- Move cassette to rack

- Move next cassette to conveyor and read ID
- Move previous cassette to conveyor and read ID
- Move current cassette to conveyor and read ID
- Move cassette to the next position
- Move cassette to a specified position
- Close detector block
- Open detector block
- Move rack down to next measurement level
22

2.1 Beginning operation of MicroBeta TriLux

Reset r(a)ck
(I)nit mask
Change (m)ask
(T)est IDs
(F)ocus ID reader
(J)ust read IDs
Print ever(y) ID
Print ra(w) IDs
Print (b)ad IDs
Si(z)e checkpoint
Calib (0)-pos
Calib det bloc(k)
(.)Park det
Shelf (1)..(16)
(*)Calib sensors

- Move rack to the highest level

- Initialize detector mask (small holes)
- Change detector mask
- Read and print ID of each cassette continuously
- Focus and stop with ID reader on
- Read IDs continuously and show IDs on the screen
- Toggle printing of ID bitmaps in ordinary form
- Toggle printing of ID bitmaps in raw form
- Toggle printing of bad ID bitmaps in raw and ordinary
form
- Move cassette to cassette type checkpoint
- Move cassette between zero point and D06
- Move detector block up and down at mask change
position
- Park detector block for shipping
- Move rack to the specified shelf
- Calibrate sensor offset steps

Operations in the Total Count Rate State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(S)how
- Show the accumulated total count rate and activity
(P)rint
- Print the accumulated total count rate and activity
(R)eset
- Reset the accumulated total count rate and activity
(D)efine efficiencies
- Define counting efficiencies of the isotopes
Operations in the System State
(H)elp
(Q)uit
MS-D(O)S
(E)rror beep
(S)tatus display
(V)ersion
(C)lock
Se(t)up mode

- Display Help text

- Back to the Ready state
- Exit to DOS
- Set error sound on or off
- Set status display on or off
- Show version number of program and MS-DOS
- Set date and time
- Select Setup mode

Setup mode contains also:

(R)S-232C
(P)rinter
(D)ata drive
Ter(m)inal

- Set serial port parameters

- Set printer parameters
- Select drive for data saving (results)
- Set terminal parameters
23

2.1 Beginning operation of MicroBeta TriLux

P(a)sswords
Customi(z)ing
(-)Detector temp
Test level contains also:
Serial (n)o
Temp cali(b)
Detector/shelf (u)sage

- Print protocol passwords

- Set environment strings to customize the program
- Set thermostat (optional)

- Set instrument serial number

- Edit temperature calibration values
- Set detector usage during counting, set program support
for the ParaLux board and set program support for the
plate ID reader if installed and specify 1/16/32 shelf
model

24

2.2 Cassettes

2.2 Cassettes
2.2.1 Construction
A MicroBeta cassette is designed to hold a microplate and to allow it to be moved from the
rack to the conveyor and back again. There are three types of cassette that can be used in
MicroBeta, a 96-well, a 24-well and a 384-well cassette. If the instrument has six or less
detectors it can take either a 96 or 24 well cassette or a 96 or 384-well cassette depending on
the mask used. If it has twelve detectors then only 96-well and 384-well cassettes can be used.

Cassette 1450-101. This has 96 sample holes with a diameter of 7.8mm. The cassette is used
when counting 1450-401 (or equivalent) 96 well sample plates.
Cassette 1450-102. This has 24 sample holes with a diameter of 13.2 mm (see the figure). This
cassette is used when counting 1450-402 (or equivalent) 24-well sample plates.
Cassette 1450-103. This has 96 sample holes. The cassette is used when counting 1450-407
plates.
Cassette 1450-104. This is a two piece cassette with 96 sample holes. This cassette is used
when counting filtermats. The filtermat in a sample bag is placed between the base plate and
the cover plate.
Cassette 1450-105. This has 96 sample holes. This cassette is used when counting 1450-410,
1450-405, 1450-419 or equivalent sample plates.
Cassette 1450-106. This has 96 sample holes. This cassette is used when counting e.g.
Millipore MultiScreen Filtration plates or equivalent sample plates.
25

2.2 Cassettes

Cassette 1450-107. This is a two piece cassette with 24 sample holes. The cassette is used
when counting Skatron filtermats cut into four pieces. The filtermat in a sample bag is placed
between the two plates of the cassette.
Cassette 1450-110. This is an open-based cassette. The cassette is used when counting 24-well
culture plates, e.g. Costar plates (or equivalent).
Cassette 1450-116. This is a two piece cassette with 24 sample holes. This cassette is used
when counting filtermats. The filtermat in a sample bag is placed between the base plate and
the cover plate.
Cassette 1450-117. This is a two piece cassette with 24 sample holes. This cassette is used
when counting 4 mL sample vials. The vials are placed between the base plate and the cover
plate.
Cassette 1450-118. This is a two piece cassette with 96 sample holes. This cassette is used
when counting e.g. 32P-labelled dot blot samples from filtermats such as 1450-423 Nylon
membrane. No scintillator needs to be added to the filtermat allowing reprobing assays. The
filtermat in a sample bag is placed between the base plate and the cover plate having solid
scintillator in sample positions.
Cassette 1450-130. This has 384 sample holes. This is used when counting 384-well sample
plates.
All the cassettes are made of white plastic with an optical surface in the sample holes.

2.2.2 Use
The sample plates are placed on top of the cassettes; the filters between the plates; the cassette
can be labelled with an ID code. This is described in chapter 2.4 "Coding the cassette".
Cassettes are loaded into MicroBeta as described in chapter 2.17 "Loading the cassette rack".
A cassette type is recognised automatically, i.e. whether it is a 384, 96 or 24 position cassette
that is being counted. See chapter 2.9 for information about which positions detectors are in
when counting samples in different types of plates.

2.2.3 Adapters and inserts

Adapter 1450-108. These adapters are used when counting Eppendorf tubes (or equivalent).
The adapters are place on a 1450-102 cassette, a bottom tape is used if needed.
Inserts 1450-109. These inserts are used when counting 24 well culture plates. The inserts are
placed in the sample wells in order to reduce crosstalk between the wells.

26

2.3 Clock settings

2.3 Clock setting

2.3.1 Current date and time
The current date and time are shown in the upper right corner of the screen during counting.

2.3.2 Adjusting the clock

When the instrument is installed you need to set the clock. Apart from this the clock will not
normally need resetting as it has a battery back-up. However if you do need to adjust the clock
proceed as follows.
Press S (System) in the Ready mode, then press C (Clock) to make changes to the clock. Enter
the date and time in the format described below.
First the program asks for the date. Type it in using the format dd-mmm-yyyy where:
dd = day of the month (1-31)
mmm = month (the first three letters of the month in English i.e JAN, FEB, MAR, APR, MAY,
JUN, JUL, AUG, SEP, OCT, NOV, DEC.)
yyyy = year (four digit year number)
E.g. 10-MAY-2000
When you have typed the date press Enter.
Note: You can also give the date in the format Year-month-day if preferred, e.g. 2000-5-10
Then the program asks for the time. Type it in using the format hh:mm:ss where:
hh = hours (0 - 23, 24 hour clock)
mm = minutes (0 - 59)
ss= seconds (0-59) (optional)
E.g. 14:57. When you have done this then press Enter.

27

2.3 Clock settings

The last line asks: Set clock. When Y (yes) is pressed the clock is started. To get back to the
Ready state press Q (quit) to quit the System mode.
For more information on the System mode see chapter 2.30 System.

2.3.3 Counting time

During counting both the elapsed time from the beginning of counting and the set counting
time are displayed on the screen (if the display is selected). If the number of cassettes is
defined the estimated end time is also displayed.

28

2.4 Coding cassettes and filtermats

2.4 Coding cassettes and filtermats

2.4.1 Cassette code system
Barcode labels are used for cassette and sample recognition. They are attached to the marked
area of the ID support plate. This is a separate piece of plastic which can be clipped onto the
appropriate area of a cassette as shown in the figure. When fixing ID labels on the ID support
plate take care that the fixing area is clean.
Sometimes it happens that the protocol ID label is dirty or placed the wrong way round and the
barcode reader fails to read it. In this case the default protocol (prot.No. 0 ) is used.

2.4.2 Cassette codes used

An ID support plate has four fields (areas) to which a barcode can be fixed. The meaning of
each field is described below. The figure shows a counting cassette. The cassette number is 1
and the counting protocol to be used is 60.

2.4.2.1 Function codes

The FUNC field can be labelled as follows:
No label - The cassette is a normal sample cassette without any special function
STD - The cassette is to be used for DPM standardization (See chapter 2.12 DPM
Standardization) The number of the standardization protocol is given in the PROT field.

29

2.4 Coding cassettes and filtermats

NORM - The cassette is to be used for CPM normalization (See chapter 2.21 CPM
Normalization) The number of the normalization protocol is given in the PROT field.
STOP - counting will stop after this cassette has been counted.
2.4.2.2 Cassette number
This is a simple number in the range 0-99 and can be output with the results. If the function
code is also a number then 100*cassette number + function code is output (in the range
0..9999).
2.4.2.3 Protocol number
This is normally a simple number in the range 0-99 (for the exception see Stop cassette below).
The type of protocol it refers to depends on what is specified in the FUNC field. If nothing is
specified in the FUNC field then it is the number of a counting protocol.
2.4.2.4 Assay protocol number
This field is for MultiCalc assay protocols only. Note that MultiCalc must be running when
using this field, otherwise the results will be ignored.
If counting is started from MultiCalc but there is no Assay code label, then the cassette will be
counted according to the other three fields but results will not be returned to MultiCalc. This
can be used if you want to run standardizations or normalizations. You could, for example,
have a normalization cassette counted first, followed by a cassette (or cassettes) labelled with
an assay protocol. The counting protocol included in the assay protocol could make use of the
normalization results just obtained. See chapters 2.20 and 2.22 for more information about
protocols.

2.4.3 Stop cassette

If you fix a STOP code to the last cassette it means that this cassette will be counted also.
However it can be tedious to remove this STOP code and stick it to another cassette when more
cassettes are loaded. To avoid this a stop cassette can be used. Stick a STOP code on both the
protocol and function code field of an empty cassette and put that cassette after the last cassette
to be counted.

2.4.4 Cassette operation

If MicroBeta is loaded with many sample cassettes that are to be counted with different
protocols, the cassettes are coded with the respective protocol number. If there is no protocol
number on the cassette, the cassette is counted with the same protocol as the previous one i.e. it

30

2.4 Coding cassettes and filtermats

is considered to be part of the same assay. To stop the counting automatically the STOP code
should be used.

2.4.5 Filtermat coding

On the lower edge of the filtermat there are two groups of twelve small circles (see the figure).
They are for marking the date. This is done by cutting or clipping off appropriate circles. The
first twelve circles are to specify the day and the second twelve the month. In the figure above,
the leftmost 9 circles stand for the numbers from 1 to 9 and the next three circles for 10, 20 and
30 respectively. E.g. the 25th day is marked by cutting off the 20 circle (the 11th circle from
the left) and the 5 circle.
The month is marked using the rightmost 12 circles, the first circle corresponding to January,
the second February etc.
The row of 7 small circles on the right-hand edge (next to sample column A12 to H12) is used
for sample identification. The circles stand for the sequence 1, 2, 4, 8, 16, 32, 64. By cutting off
appropriate combinations of circles the filtermat can be marked with a number in the range 1 to
127.

31

2.4 Coding cassettes and filtermats

2.4.6 Plate ID reader

If the optional plate ID reader is installed then it is possible to read barcode IDs from plates.
The following barcodes are supported: Codabar, Code 39, Code 128, Interleaved 2 of 5, UPL
and EAN. The ID should be fixed to the upper part of the right side.

32

2.5 Counting

2.5 Counting
2.5.1 Starting counting
First load samples as described in chapter 2.17 Loading the cassette rack. You can then either
start counting as a terminal operation (with a terminal, terminal emulator or terminal emulation
in MultiCalc) or as a MultiCalc operation. In terminal operation the counter program must be
in the Ready or Count state before you give the instruction to start counting. The options are as
follows:
2.5.1.1 Automatic counting (terminal operation)
If your cassettes have protocol ID labels, press A in the Ready or Count state. The counting
protocol to be used will be determined by the ID labels.
If your cassettes do not have ID labels, simply type the number of the protocol to be used and
press Enter. The number you enter must be within the allowed range, 0-99. Pressing ? brings a
list of protocols to select from.
If counting was started with an A-command and the first cassette has no counting protocol
code, then the first assay will be counted using the default counting protocol (protocol number
0).
Any other cassette that does not have a counting protocol number will be counted with the
protocol defined by the most recent cassette with a counting protocol number. This means that
within one assay you only need to label the first cassette. A new label is only needed to start a
new assay.
2.5.1.2 Automatic normalization or standardization (terminal operation)
If your cassettes have normalization (or standardization) protocol and function ID labels, press
A. The counting protocol to be used will be determined by the ID labels.
If your cassettes do not have ID labels, type the protocol number preceded by N in
normalization (e.g. N15) and D in DPM standardization (e.g. D15).
Typing N? or D? brings a list of normalization or standardization protocols to select from.
Normalization or standardization can be done before sample counting (See the chapters 2.12
and 2.21).

33

2.5 Counting

2.5.1.3 Automatic counting (MultiCalc)

Your cassettes must have Assay ID labels on them. Starting from the main MultiCalc menu
press F1 (=COUNTER) and then press Enter to start counting. The Assay protocol to be used
will be determined by the ID label.
If an Assay code is missing but there is a protocol code, the samples will be counted using the
counting protocol defined but results will not be returned to MultiCalc.
If counting was started from MultiCalc but the first cassette has no protocol code, then the first
assay will be counted using the default counting protocol (prot. No. 0) but results will not be
returned to MultiCalc.
Any other cassette that does not have an Assay protocol number will be counted with the assay
protocol defined by the most recent cassette with an assay protocol number. This means that
within one assay you only need to label the first cassette. A new label is only needed to start a
new assay.
2.5.1.4 Shelf number
The program will then ask for the shelf number of the first cassette. Give this number. The
default is shelf number 1.
2.5.1.5 Delayed start
Counting can also be started after a delay. This is useful for e.g. incubation. To do this press D
when starting counting. You must then enter the delay time in minutes and press Enter. The
program will display the remaining time before starting the counting. Counting can be started
immediately by pressing S or the operation can be cancelled by pressing Q.
Delay time before start (m) 60 ->
(S)tart counting (Q)uit
Time remaining: 59:58

2.5.2 During counting

The door of the counter should be properly closed during counting and it should not normally
be opened unless counting has been interrupted by pressing O. If you need to add more
cassettes then you can open the door and place the cassettes in a suitable empty place. Close
the door. Counting of those samples which were in the counting position when the door was
opened will be done again.
While counting is taking place the uncorrected CPM or CPS values are shown on the display of
the Terminal PC. These are replaced by CCPM1, DPM1 or LCPS values when the whole plate

34

2.5 Counting

has been counted. In addition there is the information from the barcodes on the cassette: assay
number, protocol number, cassette number, function codes and the number of the shelf from
which the cassette was taken. There is also the current date and time for the assay, the counting
time set in the protocol and the current counting time in seconds for the samples in the
measuring position. If there is a specified number of samples (given on line 22 in the protocol),
or normalization or standardization samples, the assay end time and date will be given also.
During counting of an Assay protocol in MultiCalc live information from the counter is not
normally shown. However by pressing the F4 key (LIVE) some information will be displayed
e.g. protocol No., current and set counting times, and positions that are counted

2.5.3 Stopping counting

Counting can be stopped anytime by just pressing O (off). The program terminates counting. If
N (Next pos) or E (Next protocol) is pressed, the program starts to count the next sample
position or the next assay respectively. If quick view is used then N stands for the next plate.
Counting of an Assay protocol in MultiCalc is stopped by pressing the F1-key. F2 and F3 are
used for counting the next position or the next assay respectively.

35

2.5 Counting

36

2.6 Counting control

2.6 Counting control

(See chapter 2J.6 for JET information)

2.6.1 Counting parameters

The counting process is controlled by the following parameters in the counting protocol:
10 Counting time
12 Precision (2 sigma)
41 Counting control
Number of repeats
Number of replicates
Number of cycles
Maximum channel counts (a hidden parameter).
48 Delay between plates

Note: Assay protocol parameters are described in chapter 2.20 MultiCalc.

A CPM normalization protocol or DPM standardization protocol does not include parameter 41
Counting control. Instead the crosstalk normalization and crosstalk standardization protocols
contain counting time for both crosstalk samples and standards, e.g.:
10 Counting time for crosstalk
11 Counting time for standards

2.6.2 Counting time

Counting will be stopped after the specified counting time (in seconds) has elapsed from the
start of the counting, unless some other conditions have already stopped it. The maximum
counting time is 999 999.9 s (more than 11 days).

2.6.3 Precision (2 sigma)

The statistical uncertainty in samples is expressed as the standard deviation or 'sigma' value
and is calculated by dividing number one by the square root of the counting value. E.g. 10000
counts gives a 'one sigma %' value of 1%. The 2 sigma % value is two times the one sigma %
value, or 2% for 10000 counts.
In MicroBeta, precision is given as 2 sigma % in a range from 0 to 99.9. Zero means that no
precision test is used.
Make sure that the counting time is set long enough if precision is wanted. E.g. two samples of
1000 and 10000 CPMs will take 10 min and 1 min respectively to achieve a 2 sigma % value
of 2%. Precision is checked once a second. The termination flag in results output will be set to

37

2.6 Counting control

'PREC' if the precision has been reached. The counting stops if the precision has been reached
in all detectors.

2.6.4 Number of repeats

To check sample stability or to check instrument performance, the same sample can be counted
repeatedly, up to 99 times. If the repeat value is less than or equal to 5 then results are sorted
and some statistical values are calculated and printed after the sample results.

2.6.5 Number of replicates

To evaluate sample preparation errors, a number of replicates of a sample can be used. The
results for each sample are output separately. After each group of replicate samples, some
statistical values will be sent to output. The maximum number of replicates are 99.

2.6.6 Number of cycles

All the sample cassettes in the rack can be counted in repeating cycles. One cycle means
counting of all cassettes in a batch once (i.e. those being counted with one protocol). A batch
ends when the next cassette with a protocol ID is found. When the instrument has counted all
the cassettes in this batch, it moves the rack up until the first cassette in the batch is found.
Then it recounts the cassettes in the batch. Counting of the next batch starts when the cassettes
have been counted the number of times specified by the Cycle parameter. The repeating cycles
can be used to check sample stability. The maximum number of cycles are 99.
If the number of cycles is greater than 1, then you will be asked to give the Cycle delay. This is
in minutes and is the time between when one cycle ends and the next one begins.

2.6.7 Order of operation of repeated counting

If you have set two or all three of the repeat, replicate and cycle parameters to values greater
than 1 the order of operation is:
First, repeat counting of each sample. The output consists of the individual results for each
repeat count for a sample followed by statistics for the repeat.
Second, statistics for all the replicates of the sample are calculated using the statistics for the
repeat.

38

2.6 Counting control

This procedure is repeated for all samples in an assay batch. When the end of the batch is
reached, counting starts from the beginning by counting the repeats and replicates again. This
procedure continues for as many times as you have defined for the cycle parameter.

2.6.8 Delay between plates

This parameter can be used to control the interval between when the counting of one sample
plate has finished and the next is started. The range is 0 to 9999 minutes.

39

2.6 Counting control

40

2.7 Crosstalk correction

2.7 Crosstalk correction

2.7.1 Introduction
Crosstalk is a situation in which light pulses from adjacent samples interfere with the pulses of
the sample under measurement. This may occur in the case of microtitration plates in which you
cannot use the cassettes 1450-101 or 1450-102. This is because designs of cassette other than the
1450-101 or 1450-102 have part of the optical shielding between the wells removed to
accommodate the microtitration plate.
Crosstalk can be corrected with the MicroBeta program. Before counting the actual samples, the
amount and type of crosstalk is defined using special standardization or normalization samples,
with the type of solution as similar to the actual samples as possible. The normalization or
standardization is done first and the calculated crosstalk factors are then used when counting the
actual samples. A CPM normalization run is needed for crosstalk corrected CPM results and a
DPM standardization run is needed for crosstalk corrected DPM results.

2.7.2 Crosstalk CPM normalization

2.7.2.1 Preparation of normalization samples on a 96 or 384-well microtitration
plate
Take sample solution with some activity (+ scintillant) which corresponds to the samples to be
analyzed, i.e. has the same isotope and solvent, and pipette the same volume as in the samples,
into well G11 (96-well plate) or well N22 (384-well, 1-6 detectors) or well M21 (384-well, 12
detectors).
For a 96-well plate with a 1-6 detector counter, pipette background solution (corresponding to the
sample solution, but without activity) into wells A1, F11 and G10 (96-well), where the latter two
are the crosstalk samples. G10 is only needed for strip plates and A1 if background correction
has been selected. With a 12 detector counter use positions A1 and F11. Strip plate is not
available. For a 384-well plate with a 1-6 detector counter use positions A1, M22 and N21 for
strip plate. With a 12 detector counter use positions A1 and M22. Strip plate is not available.
The sample array for a 96-well plate with a 1-6 detector counter is as follows, where S1 is the
sample solution and B1, C1 and C2 are the background solutions:

41

2.7 Crosstalk correction

A
B
C
D
E
F
G
H

10

11

C2

C1
S1

12

Close the sample plate with a sealing tape and place the plate on a 1450-103 or 1450-105 cassette
for counting.
2.7.2.2 Preparation of normalization samples on a 24-well microtitration plate
The procedure is the same as with 96-well plates, but the normalization sample positions are D6
for the sample solution and A1, C6 and D5 for the background solution. D5 is only needed for
strip plates; A1 is needed if background correction has been selected. The sample array is as
follows:
1

A
B
C
D

C2

C1
S1

2.7.2.3. Normalization Protocol

Edit a normalization protocol for crosstalk. To do this in the Ready-state press P (P)rotocols then
N (N)ormalization protocols.
Select (E)dit and a protocol number.
Edit the protocol parameters, see the figure overleaf for an example of the parameters.

42

2.7 Crosstalk correction

1 Protocol name:
crosstalk correction
3 Crosstalk correction (Y/N)
Y
5 Isotope 1:
1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
1
9)Luminescence
PMT use: 1)Normal 2)Upper 3)Lower
1
10 Counting time for crosstalk [s]
60.0
11 Counting time for standards [s]
60.0
12 Precision (2 sigma) [%]
0.2
30 Printer output: 1)No
2)Short
3)Long 4)Programmable
3
32 Display output: 1)No
2)Short
3)Long 4)Programmable
3
34 External output: 1)No
2)Short
3)Long 4)Programmable
1
36 File output:
1)No
2)Short
3)Long 4)Programmable
1
40 Change special features (Y/N)
Y
41
Isotope activity setting (Y/N)
N
42
Background sample (Y/N)
N
43
Half-life correction (Y/N)
N
44
Chemiluminescence correction (Y/N)
N
45
Use password (Y/N)
N
46
Special plate: 1)Wallac 2)BP filter
3)Other
1
47
Strip plate (Y/N)
N

Select crosstalk correction on line 3, as well as the isotope (line 5) and counting times (lines 10
and 11). You can give the isotope activity setting on line 41 (Y for yes, and then the DPM value).
A background sample is counted only if selected on line 42. If you use strip plates then select Y
on line 47. The crosstalk sample in G10, D5 or N21 is counted only if strip plate is selected.
2.7.2.4 Normalization counting
Start the normalization run in the Ready or Count state using automatic counting and ID codes,
or by selecting Nx, where x is the number of the protocol.
The normalization sample is then counted in every detector and the crosstalk sample in detector
1.
2.7.2.5 Counting the actual samples using crosstalk correction
Edit a counting protocol:
Select in the Ready-state P (P)rotocol then C (C)ounting protocols.
Then select E ((E)dit) and a protocol number.

43

2.7 Crosstalk correction

On parameter line 2 select 1 (CPM) and on line 3 select the number of the crosstalk
normalization protocol defined above. Edit the other parameters normally.
Start counting using automatic counting and a protocol ID label or start with the protocol number
and press Enter.
When counting the samples, the stored crosstalk factors are used for correcting the CPM values.
The crosstalk corrected CPM values are marked as CCPM1 in the printout.

2.7.3 Crosstalk DPM standardization

2.7.3.1 Preparation of DPM standardization samples on a 96 microtitration plate
(1-6 detector counter)
Make first the standard samples in vials and then pipette them onto a plate:
a) 6 vials for isotope standard samples and 6 vials for blank (i.e. crosstalk) samples are needed.
b) Add into each standard sample vial (numbered from 1 to 6) scintillation liquid and isotope
solution, or pipette the isotope solution directly into the sample wells, the same amount in
each. Add the same solution into blank vials/wells, but without the isotope. (If there is no
isotope in the standard vials, the same vials can be used for blank samples.)
c) Add quencher into each standard and blank vial, so that the samples form a quench series, i.e.
the amount of quencher increases from vials 1 to 6. The amount of quencher added (here
CCl4 is used as a quencher) forms a series as follows:
Vial No. Amount of CCl4/5 ml
l
1
0
2
5
3
15
4
30
5
50
6
75
Shake the vials. Then pipette the standards and blanks onto a sample plate according to the
following array (use the same total volume in standards and blanks as in the actual samples):

44

2.7 Crosstalk correction

A
B
C
D
E
F
G
H

1
C1
S1

3
C2
S2

5
C3
S3

7
C4
S4

9
C5
S5

10

11
C6
S6

S1
C1

12

S6
C6

The isotope standard samples are marked with S and a number, which refers to the vial number
(the amount of quencher). The blank samples are marked with C and a number.
Close the sample plate with a sealing tape and place it on a 1450-103 counting cassette.
2.7.3.2 Preparation of DPM standardization samples on a 24-well microtitration
plate
The procedure is similar to that described above but two plates are used and the sample positions
are as follows:
First plate

A
B
C
D

1
C1
S1

3
C2
S2

5
C3
S3

S1

C1

5
C6
S6

S6

C6

Second plate

A
B
C
D

1
C4
S4

3
C5
S5

45

2.7 Crosstalk correction

2.7.3.3 DPM standardization protocol

Make a DPM standardization protocol by selecting in the Ready-state P (P)rotocols then S
(S)tandardization protocols.
Select edit (E) and a protocol number.
Edit the protocol, see the figure below.
Line (1..40, type / to exit, ? for help)
1
1 Protocol name:
->
2 Sample type: 1)Normal 2)SPA
1
3 Crosstalk correction (Y/N)
N
correction
5 Isotope 1: 1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
1
PMT use: 1)Normal 2)Upper 3)Lower
1
Window 1:
5- 320
10 Counting time for crosstalk [s]
60.0
11 Counting time for standards [s]
60.0
12 Precision (2 sigma) [%]
0.2
20 Number of standards
6
21 Isotope 1 activity [DPM]
200000.0
24 Standard curve fit selection
25

26
30
32
34
36
40

(Y/N)

Curve fit method: 1)Smoothing spline

2)Interpolation spline
3)Linear interpolation
4)Linear regression
Automatic smoothing (Y/N)
Edit standard curve (Y/N)
Printer output: 1)No
2)Short
3)Long 4)Programmable
Display output: 1)No
2)Short
3)Long 4)Programmable
External output: 1)No
2)Short
3)Long 4)Programmable
File output:
1)No
2)Short
3)Long 4)Programmable
Change special features (Y/N)

->
->
->Y Select crosstalk

->
->
->
->
->
->
->

Activity of standards is
given on line 21
N ->Y Curve fitting method
given here

1 ->
Y ->
N ->
3 ->
3 ->
1 ->
1 ->
N ->

Select crosstalk correction on line 3. Remember also to edit the parameters: isotope, counting
time, number of standards and the isotope activity. See the listing above.
2.7.3.4 DPM Standardization Counting
Start the DPM standardization run in the Ready-state or Count-state using automatic counting
and ID-labels, or by selecting Dx, where x is the protocol number (no ID-label is needed in this
case).

46

2.7 Crosstalk correction

The active samples are automatically counted first, e.g. positions G9, B1-B11 and G12, then the
crosstalk samples A1-A11, H9 and H12.
2.7.3.5. Counting the actual samples with crosstalk correction
In the Ready-state select a counting protocol to edit.
On parameter line 2 select DPM-mode and on line 3 the crosstalk DPM standardization protocol
defined above.
Start automatic counting using ID-labels or start with the protocol number and press Enter.
The DPM values in the printout are the crosstalk corrected values.

47

2.7 Crosstalk correction

48

2.8 Datafiles

2.8 Datafiles
2.8.1 Use
Counting results can be stored on a floppy or hard disk located in the PC (terminal emulation
or MultiCalc); they can also be sent to the server of a local area network (LAN). Results can
also be stored on the MicroBeta disks in drive A: or B:. If a VT52 or VT100 terminal is used
then results are stored on the protocol disk in MicroBeta drive B:. (see 2.31.6.4 Terminal). The
data in the files saved on the disk is available to be processed or printed.
Data can also be sent to an external computer. The data is sent via port 2 on the back of the
counter.
Note: Port 1 is for the Terminal PC. The third output, port 3, is for the printer.

2.8.2 Disk drive selection

The disk drive of the terminal PC where the data is to go can be chosen in the System state by
pressing D (Data drive). You can choose from drives A, B or C where C is a hard disk drive
and A and B are for floppy disks. You can also specify the whole directory path (see the
chapter System/(D)ata drive). Start the path with @ if you wish to store results on the
MicroBeta disk.
Depending on the type of PC you have to proceed as follows:
Single floppy disk PC - You must select drive A. Then you must replace the terminal emulator
program diskette with an empty but formatted data diskette.
Dual floppy disk PC - Select drive B. This allows you to keep your terminal emulator diskette
in drive A. Load an empty formatted data diskette into drive B.
Hard disk PC - Select drive C (hard disk) or specify the path. This will allow you to collect
large amounts of data. If you have only specified the drive you must select the directory on the
terminal PC where you want to store your data. To do this, exit from the terminal emulator
program. (See the chapter Terminal emulators/Exit to MS-DOS). Use the change directory
command in MS-DOS i.e. cd\directory name to go to the directory you want the data to be
stored in. When you have done this, then return to the terminal emulator program by typing
EXIT and pressing Enter.

49

2.8 Datafiles

The disk drive and the directory for the results files can also be selected in the protocol
together with the output file selection. This will override the selection made in the System state
and will make it possible for users to have their own results files directories
When you have selected the directory you can instruct the program to save data on it as
follows.

2.8.3 Saving data in datafiles.

Select protocol operations by typing P, N or S when in the Protocols state. (P is for counting
protocol, N for normalization protocol and S for DPM standardization protocol).
Next choose the protocol to be edited by giving the group number and pressing Enter.
You can edit any parameters in the protocol but in particular go to line 36 File output and select
which data you want to save (see chapter 2.22 Protocols for details). The example shows that
the Long output has been selected. Then on the line File path enter the disk drive and directory
or leave it as it is if the drive defined in the System/Data drive shell is used. In this example a
subdirectory TEST belonging to directory MB on PC disk A has been selected.
36 File output:

1)No
3)Long

File path:
File name extension
40 Change special features

2)Short
4)Programmable
1 ->3
C:\MB\->A:\MB\TEST\
1 ->
(Y/N)
N ->

Before starting counting be sure that you have inserted an empty formatted data diskette into
the drive selected or, if you have selected saving to hard disk (drive C:), be sure that the current
directory has been selected as described in the previous section. If the file path is given, then
this directory does not need to be the current directory, but it must exist. Use the MKDIR
command in MS-DOS to make new directories. Format new diskettes by the MS-DOS
FORMAT command.
When you have finished protocol editing, store the protocol by pressing / and begin counting
by pressing A in the Ready or Count state.
Note: Line 36 is not available if UltroTerm is used. The filing system of UltroTerm should be
used in this case.

2.8.4 Accessing datafiles

Result files that are stored on the data disk will be named so that the first part of each name
(the part before the point) is the protocol number and the second part (after the point) is a
50

2.8 Datafiles

running number indicating the assay execution order. E.g. for protocol 5 the first saved file is
named 5.001, the second 5.002 etc. Other files have the format e.g. S5.001 for standardization
files and N5.001 for normalization files. The file name extension can be changed in the
protocol. To access a file on the data disk in the terminal PC, exit from the terminal emulator
program to MS-DOS (see the chapter Terminal emulators/Exit to MS-DOS).

2.8.5 External datafiles

Results can be sent to an external data collection device by connecting it to port 2. The external
output and the result format are chosen on protocol line 34 External output.
34 External output: 1)No
2)Short
3)Long 4)Programmable
35 External output: POS CCPM1 CCPM1%
->POS CTIME CPM1 CPM1% CCPM! CCPM1%

1 ->4

There are four alternatives which you can select on protocol line 34 External Output:
1. Nothing is sent
2. Only position number, CCPMs and CCPM%, DPM and DPM%, or LCPS, LCPS% and
FLAG are sent (CCPM is corrected CPM and % means the percentage error in the CCPM,
DPM or LCPS value). CCPM, DPM or LCPS are also sent in PLATE format
3. The long results printout including SQP(I) and CPMs is sent (see the Results chapter)
4. User selected outputs are sent

2.8.6 Data processing

Counting results are stored in ASCII format, and they can be processed using commercial
spreadsheet programs.

2.8.7 Datafiles in MultiCalc

Results from MultiCalc controlled Assay protocols are handled in a different way. The types of
results files are selected in the Assay protocol editor. Input is an internal file for saving data to
be used later in MultiCalc, e.g. in evaluations. Results is a text file to be used mainly for
transferring data to an external computer. See the User Guide to MultiCalc functions manual
and the module called File handling and evaluation.

51

2.8 Datafiles

Results from MicroBeta controlled protocols are handled as with any other terminal emulator,
i.e. they are sent via MultiCalc to the drives and directories specified in the protocols. These
result files cannot be evaluated by MultiCalc.

52

2.9 Detectors

2.9 Detectors
2.9.1 Six detector model
This model of MicroBeta has twelve photomultiplier tubes arranged in six pairs. Each pair
works in coincidence. The tubes of the pair are situated on opposite sides of the counting block,
one above and one below the block. The detectors form a 3 x 2 array, see the figure below.
Six samples are counted simultaneously, which reduces counting time to about one sixth of that
for a single detector instrument.
When a cassette is being counted, the transportation system moves it between the detector
blocks. The first detector then counts position A1, the second A5 (A3 for 24-well plates), the
third A9 (A5), the fourth E1 (C1), etc. (see the figure below). At this phase only the results
from position A1 are output, because the results must be in the order A1, A2, A3, etc. The
cassette moves one position forward and positions A2, A6, A10, E2, E6, and E10 are counted.
The results from A2 are output. After counting the whole row the cassette moves to the
beginning of the next row (e.g. position B1 in detector 1). The rest of the results from row A
are output.

Six detector
model, cassette in
first position

Only the first detector can count every position in a plate. The following table shows which
positions can be counted by each detector:

53

2.9 Detectors

Det.No.
1

96-well
every

24-well
every

384-well
every

A5-A12
......
H5-H12

A3-A6
.....
D3-D6

A9-A24
......
P9-P24

A9-A12
......
H9-H12

A5-A6
.....
D5-D6

A17-A24
.......
P17-P24

E1-E12
......
H1-H12

C1-C6
D1-D6

I1-I24
......
P1-P24

E5-E12
......
H5-H12

C3-C6
D3-D6

I9-I24
......
P9-P24

E9-E12
......
H9-H12

C5-C6
D5-D6

I17-I24
.......
P17-P24

The smallest range of positions is that counted by detector 6. This is therefore the only range
that can be counted by all detectors. This affects the positioning of CPM normalization and
DPM standardization samples because these routines involve every detector counting the same
sample. See chapters 2.12 and 2.23 for details.

54

2.9 Detectors

2.9.2 Three, two and one detector models

The MicroBeta series also includes models with 3, 2 and 1 detectors. The detectors are then
arranged as shown in the following figures.

Three detector
model, cassette in
first position

Two detector
model, cassette in
first position

One detector
model, cassette in
first position

55

2.9 Detectors

2.9.3 Selecting detector number

The amount of detectors must be defined in the system parameters before starting counting. Go
to the System state by pressing S and select the test level by pressing L followed by T. Then
select detector usage by pressing U and choose one of the alternatives for the number of
detectors. Return to the normal level by pressing L followed by N before quitting to the Ready
state by pressing Q.

2.9.4 Twelve detector model

A twelve detector MicroBeta has twenty-four photomultiplier tubes arranged in twelve pairs.
Each pair works in coincidence. The tubes of the pair are situated on opposite sides of the
counting block, one above and one below the block. The detectors form a 4 x 3 array, see the
figure below.

Twelve detector
model, cassette in
first position

Twelve samples are counted simultaneously, which reduces counting time to about one twelfth
of that for a single detector instrument. There are nine readings per plate.
When a cassette is being counted, the transportation system moves it between the detector
blocks. The first detector then counts position A1, the second A4 etc. At this phase only the
results from position A1 are output, because the results must be in the order A1, A2, A3, etc.
The cassette moves one position forward and positions A2, A5 etc. are counted. The results
from A2 are output. After counting the whole row the cassette moves to the beginning of the
next row (e.g. position B1 in detector 1). The rest of the results from row A are output.
A limited range of positions (10 to 12 in row G) can be counted by all detectors. This affects
the positioning of CPM normalization and DPM standardization samples because these
routines involve every detector counting the same sample. See chapters 2.12 and 2.23 for
details.

56

2.10 Diskettes

2.10 Diskettes
2.10.1 Diskettes used
The diskettes used in the MicroBeta counter itself are 3.5" micro-floppy disks with standard
1.44 MB format. The program diskette has to be in drive A (the lower drive) and the protocol
diskette in drive B (the upper drive).
A diskette is inserted into a disk drive with the label upwards and towards you. When the
diskette is properly in its place, a click is heard. To remove the diskette press the button under
the diskette slot.
Terminal emulator diskettes are normally 3.5" (1.44 MB) micro-floppy disks and these are
inserted into the A drive of the terminal PC.

2.10.2 Diskette handling and storing

A floppy disk should be handled carefully. Avoid touching the magnetic surface, and always
when handling the diskette touch only the end with the label on. Do not bend the diskette, or
write on it. When labelling it the label should be written first and affixed afterwards on the
diskette. Do not place anything heavy on it (e.g. books). Put the diskette in a cardboard
envelope when not in use. Diskettes should be stored in a closed box to avoid dust getting on
them. The storage temperature should be between 10 - 30o C. They should also be kept away
from magnets.

57

2.10 Diskettes

2.10.3 Backup of the terminal diskette

To make a backup copy of the terminal program or data diskette in the terminal PC use the
MS-DOS DISKCOPY command (see 2.31.2.8 Terminal emulators/ Backup).

2.10.4 Backup of the program diskette

There should always be a backup copy of the program diskette in case something should
happen to the original. To make a backup copy, start the MicroBeta program as normal. Then
type S to get to the System state and there type O for exit to MS-DOS. Press Y twice to
confirm the exit. After the prompt MicroBetaA:\> appears on the screen type PROGRAM, and
press Enter. Follow the instructions on the screen. Put a new diskette into drive B: so that a
copy can be made. Store the new copy of the program diskette in a safe place.
To get back to the MicroBeta program put the protocol diskette back into drive B, type MB and
press Enter.

2.10.5 Backup of the protocol diskette

Protocol modifications are saved on the protocol diskettes so it is advisable to make a backup
copy of these diskettes from time to time in order to save the protocols used.
To make a backup copy of the protocol diskette, begin in the same way as with the program
diskette. The procedure differs from the point when the prompt MicroBeta A:\> is displayed.
Here type PROTOCOL BACKUP and press the Enter key. Remove the program diskette and
put a new protocol diskette into drive A to receive the protocols.
If you want to make a new empty protocol diskette, then type PROTOCOL NEW instead, and
press the Enter key. Then follow the instructions displayed on the screen.
Now store the copied diskette as a backup copy, and use the other diskette as a protocol
diskette in MicroBeta, or, if a new empty protocol diskette was made, use it for saving new
protocols.
To get back to the MicroBeta program, put the program diskette back into drive A, type MB
and press Enter.

58

2.10 Diskettes

2.10.6 Preparing a 3.5 protocol diskette

A standard 3.5" MS-DOS 1.44 MB formatted diskette can be used as a MicroBeta protocol
diskette. When you want to initialize an empty formatted diskette to be used as a protocol
diskette, insert the empty diskette into drive B on the counter and start up MicroBeta. When the
program has been loaded, the following message should appear:
Cannot find the protocol index

Press M. The program will try to find the protocols on the diskette and will create the protocol
index. Then the program returns to the Ready state and the protocol diskette is ready for use.

59

2.10 Diskettes

60

2.11 DPM counting

2.11 DPM counting

2.11.1 Why DPM counting?
CPM counting is used when sample preparation is expected to yield samples with close to
constant counting efficiency, i.e. no variation in quench level. This means that the results of the
samples in an assay can be compared with each other and used in further data analysis. If the
counting efficiency varies with quench in the sample, CPM counting can no longer give
accurate results, DPM counting is therefore needed.
DPM counting involves the use of quench curves. These are made after counting a number of
standards with different quench levels. This procedure is called DPM standardization. The
spectra from counting unknown samples are compared with the quench curve and accurate
results are calculated.
In MicroBeta the quench curves are constructed by first counting 2 to 12 DPM standards in
detector 1 and then counting two so called DPM normalization standards in every detector. The
curve from detector 1 is recalculated for the other detectors depending on the detector
efficiencies. See chapter 2.12 DPM standardization for more information about the
standardization procedure.

2.11.2 Single label DPM

The DPM standards are counted in detector 1. MicroBeta has no external standard so the
spectrum quench parameter for the isotope (SQP(I) ), or the Asymmetric quench parameter for
the isotope, AQP(I), is therefore used for determining the quench curve. AQP(I) is used when
ParaLux counting is selected. The normalization standards are then counted in every detector in
order to get the efficiencies and SQP(I) (or AQP(I)) values.
Later, when measuring an unknown sample, the CPM and SQP(I) (or AQP(I)) values are
counted in a particular detector. SQP(I) (or AQP(I)) is modified to fit as SQP(I) (or AQP(I))
for detector 1. The efficiency for detector 1 is counted from this SQP(I) (or AQP(I)) by using
the quench curve, and is then used for determining the efficiency for the particular detector.
The DPM value is counted by dividing CPM with the efficiency. See chapter 3.3 Calculation
methods for formulae used in DPM counting.

2.11.3 Easy DPM

Easy DPM is a special case of single label DPM counting. Only normalization standards are
counted in standardization and stored quench curves are used when counting unknown
samples.

61

2.11 DPM counting

2.11.4 Dual label DPM

Dual label counting is similar to single label counting except that two plates are used, one for
each isotope. The DPM1 and DPM2 values are calculated using the 3/2 method.
Note: EasyDPM is not available with ParaLux count mode.

62

2.12 DPM standardization

2.12 DPM Standardization

2.12.1 What is standardization?
When counting DPM samples in a multidetector counter the variation in detector efficiency,
detector energy and the effect of quenching must both be corrected for. Standardization is the
name given to the process in MicroBeta which makes this correction.
2.12.1.1 Quench correction
Several samples labelled with the same activity will give different count results even in the
same detector because of the difference in the amount of energy absorbed in the samples. This
is called quenching. To correct for this a number of samples with identical DPM but differing
quench have to be counted in one detector. This establishes a relationship between a quantity
called SQP(I) or AQP(I) (the latter is used for ParaLux counting) and efficiency. SQP(I) (or
AQP(I)) is a measure of the isotope spectrum and varies as the quench changes. Once this
relationship between SQP(I) (or AQP(I)) and efficiency is known it is possible to calculate the
counting efficiency for any measured SQP(I) (or AQP(I)) value in the range covered by the
measurement of the standard samples. Knowing the efficiency, the DPM can be calculated.
2.12.1.2 Detector energy and efficiency calibration
However closely matched detectors are, there will be some difference in their energy
calibration (affecting SQP(I) (or AQP(I)) measurement) and counting efficiency. A correction
must be made for both these differences.
2.12.1.3 Standardization
Standardization = Detector energy calibration + Sample quench correction + detector
efficiency calibration
In MicroBeta the standardization is done by first measuring a maximum of 12 DPM standards
with defined activities in detector 1 and then counting two DPM standards in every detector.
The two calibration standards should be representative of the range covered by the DPM
standards, i.e. they could be replicates of the first and last DPM standards.
The quench standard measurement produces a curve of efficiency versus SQP(I) (or AQP(I))
with as many points as standardization samples.
Energy calibration and efficiency calibration are both linear functions so two points are
sufficient to define these two calibration curves. The two points are obtained from the counting
of the two calibration standards mentioned earlier.

63

2.12 DPM standardization

It is preferable that each standard is counted with the same amount of pulses to get an accurate
curve. Set the counting precision on line 12 of the counting protocol. Make sure the counting
time is long enough that the precision requirement will terminate counting and not the counting
time. See chapter 2.25 Protocols section 2.25.7 number 12 for more details.
2.12.1.4 Easy DPM
If Easy DPM is selected, then DPM standards are not needed. The instrument uses stored
quench curves for isotopes 3H, 125I, 14C, 35S, and 32P. Only two normalization samples are
measured in standardization. Normalization sample positions are H11 and H12 (96-well plate
1-6 detectors), G11 and G12 (96-well plate, 12 detectors), D5 and D6 (24-well plate), P23 and
P24 (384-well plate, 1-6 detectors) M22 and M23 (384-well plate, 12 detectors).
2.12.1.5 Evaluating unknown samples
The actual sequence of events for counting unknown DPM samples is as follows:
1 The SQP(I) (or AQP(I)) value and CPM are obtained with any detector.
2 The SQP(I) (or AQP(I)) value is now corrected from the energy calibration curve to be what
it would have been if the sample had been counted in detector 1.

3 The corrected SQP(I) (or AQP(I)) is used to read off the detector 1 efficiency from the
quench curve made for samples counted with detector 1, see the figure overleaf.

64

2.12 DPM standardization

4 The efficiency calibration curve is then used to obtain the actual detector efficiency
corresponding to the detector 1 efficiency just obtained.

5 The measured CPM value is then divided by the actual detector efficiency to obtain the
activity in DPM.

2.12.2 Editing standardization parameters

Standardization parameters are stored in standardization protocols (see the listing in section
2.12.3). There can be a maximum of 100 standardization protocols, numbered from 0 to 99.
The default protocol is number 0.
Protocols 91 to 95 , 97 to 98 are made at the factory for isotopes 3H, 125I, 14C, 35S, 32P and 32P
Cerenkov respectively. These may not be deleted because they are connected to the
corresponding counting protocols. Note that no standardization has been done for those
protocols i.e. there is no standardization data connected to the protocols. Standardizations must
actually be performed with the protocols to get the standardization data.
Press P and S in the Ready state to choose the standardization protocol mode. The protocols
can then be edited, copied, deleted, printed and displayed, and the curves can be plotted.

65

2.12 DPM standardization

Press the appropriate key for the following options:

F - default parameters
E - edit a protocol
C - copying a protocol including standardization data
D - deleting a protocol
Note: the standardization data will be destroyed when editing an old standardization
protocol. The standardization must therefore be done again for this protocol. Only the
standardization protocol name, curve fitting method, the curve and password can be edited
without destroying the standardization data.
See chapter 2.25 Protocols for further details on standardization parameters and editing.

2.12.3 Preparing single label standardization samples

Two to 12 DPM standards are placed in positions A1 to A12 in a 96 or 384-well plate or in
positions A1 to B6 in a 24-well plate. No empty positions in the beginning or between samples
are allowed. The first and last normalization standards must be also in position H11 and H12
(96-well, 1-6 detectors), G11 and G12 (96-well, 12 detectors), D5 and D6 (24-well), P23 and
P24 (384-well, 1-6 det.) and M22 and M23 (384-well, 12 det.). If Easy DPM is selected then
only normalization standards are needed.

66

2.12 DPM standardization

If ParaLux counting is selected, then it is possible to subtract thermal background from the
AQP(I) and counts values. Select Background sample on line 42 and put an empty cassette as
the first standardization cassette followed by the usual standardization cassette.
Line (1..40, type / to exit, ? for help)
1
1 Protocol name:
->
2 Sample type: 1)Normal 2)SPA
1
3 Crosstalk correction (Y/N)
N
4 Number of labels: 1)Single 2)Dual
1
5 Isotope 1: 1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
9)Luminescence
1
ParaLux (Y/N)
Y
ParaLux mode: 1) High efficiency
2) Low background
2
Discriminator channel
150
PMT use: 1)Normal 2)Upper 3)Lower
1
Window 1:
5- 360
10 Counting time for crosstalk [s]
60.0
12 Precision (2 sigma) [%]
0.2
14 Easy DPM (Y/N)
N
20 Number of standards

6
21 Isotope 1 activity [DPM]
200000.0
24 Standard curve fit selection
25

26
30
32
34
36
40
41
42

43
44
45
46
47

(Y/N)

->
->
->

->
->
->
->
->
->
->
->
->
->

Activity of standards is
given on line 21
N ->Y Curve fitting method
given here

Curve fit method: 1)Smoothing spline

2)Interpolation spline
3)Linear interpolation
4)Linear regression
1
Automatic smoothing (Y/N)
Y
Edit standard curve (Y/N)
N
Printer output: 1)No
2)Short
3)Long 4)Programmable
3
Display output: 1)No
2)Short
3)Long 4)Programmable
3
External output: 1)No
2)Short
3)Long 4)Programmable
1
File output:
1)No
2)Short
3)Long 4)Programmable
1
Change special features (Y/N)
N
Isotope activity setting (Y/N)
N
Background correction (Y/N)
N
Use normalization background (Y/N)
N
Background sample (Y/N)
N
Bgnd sample position
G12
Bgnd counting time [s]
60
Half-life correction (Y/N)
N
Chemiluminescence correction (Y/N)
N
Use password (Y/N)
N
Special plate: 1)Wallac 2)BP filter
3)Other
1
Strip plate (Y/N)
N

67

->

->
->
->
->
->
->
->
->Y
->
->Y
->
->Y
->
->
->
->
->
->
->

Background sample
selected

2.12 DPM standardization

2.12.4 Preparing dual label standardization samples

Two plates are used and the higher energy isotope samples are counted first. A maximum of 6
DPM standards can be used on each plate. For 24-well plates the sample positions are the same
as in single label standardization.
For a 96-well plate the standards are placed in the following positions on each plate: B1, B3, ..
, B11, the first normalization standard in position G9 (96-well, 1-6 det.), G10 (96-well, 12 det.)
and the last normalization standard in position G12 (96-well 1-12 det.).

2.12.5 DPM standardization procedure

The plate(s) with the standardization samples are placed in cassette(s) which are identified with
the corresponding standardization protocol number in the protocol number area and with a
STD label in the function code area. In dual label standardization the first (higher energy
isotope) cassette should have the STD label or STD2 for the first plate and STD1 for the
second. Insert the cassette(s) into the rack and close the door. Press either A (Automatic
counting) or Dxx, where xx is the standardization protocol number, in the Ready or Count state
to start counting. If Dxx is selected the program asks:
Do you want to continue counting after standardization? (Y/N)->

If Y (Yes) is selected the counting continues after counting the standardization plate, if N (No)
is selected the counting stops after the standardization.
If standardization has already been done for the protocol, then the program asks first:
This protocol contains standardization data. Continue? (Y/N) ->

This is to prevent accidental overwriting of standardization data.

If automatic counting is selected the standardization cassette can be placed anywhere in the
rack. The STD label is needed in this case.
The counting results for the standardization are output as specified on lines 30 - 37 in the
standardization protocol.

2.12.5 Standardization results

When a standardization is done the results are stored with the standardization protocol. The
standardization data can be used by one or several counting protocols and is selected when
editing the protocol (see chapter 2.25 Protocols).

68

2.13 Errors

2.13 Errors
2.13.1 Program failure
If the program is jammed and will not take any orders typed on the keyboard, it is best to
restart the program. Turn the power off from the counter and the terminal. Switch the power on
again, first the terminal and then the counter and try again. If the counter does not restart then
remove the battery fuse before switching the power on.

2.13.2 Pressing the wrong keys

If a wrong key is pressed, the beep is heard and alternatives are shown again. If in protocol
editing a wrong value is given, it can be easily erased by pressing the Backspace key and
retyping the correct value. If the Enter key has already been pressed, then type L and the line
number to return to the line to enter a new value or either the up arrow or P and Enter to go to
the previous line.

69

2.13 Errors

70

2.14 Half-life

2.14 Half-life
2.14.1 Radioactive half-life
The half-life of a radioactive isotope is the time in which half of the radioactive nuclei have
decayed, i.e. the activity is halved. The decrease in activity is exponential and can be calculated
using the formula:
A(1) = A(0)x2(-t/Thalf)
Where: A(1) = the activity at time t(1), A(0) = the activity at time t(0) and t=t(1) - t(0). Thalf =
the half-life of the isotope.
Each radioactive isotope has its specific half-life. If the half-life of an isotope to be counted is
short, and if the counting time of a batch of samples is relatively long, the last samples will
have decayed more than the first ones so the results between earlier and later samples cannot be
directly compared with each other. As an example: the half-life of the 32P isotope is 14.3 days,
and if counting takes 24 hours, the last samples will give about 5 % lower activity than they
would have done if counted at the same time as the first sample.

2.14.2 Half-life correction

The half-life correction is calculated, if requested, during protocol setting. This correction is
recommended if the half-life of the isotope used is short and/or the total counting time is long.
If you select one of the six defined isotopes in protocol setting, the half-life will appear on line
43, see the figure.
40 Change special features (Y/N)
N
41
Isotope activity setting (Y/N)
N
42
Background sample (Y/N)
N
43
Half-life correction (Y/N)
N
Half-life 1 (h)
107500.00
Zero time 1 set (Y/N)
N
Zero date 1
28-Jun-2000
Zero time 1
15:40:53
44
Chemiluminescence correction (Y/N)
N
45
Use password (Y/N)
N
46
Special plate: 1)Wallac 2)BP filter
3)Other
1

->Y
->
->
->Y
->
->
->
->18:30
->
->

Tritium was the isotope

selected on line 5 and the
half-life is given
automatically after line 43

->

If you select isotope option 7 "Other" then you must specify the half-life in hours on line 43. If
you type "0.0" no half-life correction will occur. The next line asks if you want to set a zero
time. If you select Y then two more protocol lines appear to allow you to set the zero date and

71

2.14 Half-life

zero time. This corresponds to time t(0) described earlier in the formula. This is the moment
back to which all results are corrected. If you select N to the zero time setting question then
activities are corrected to the time when counting of the batch of samples was started.
Isotope
3
H
14
C
32
P
35
S
51
Cr
125
I

Half-life
12.3 years
5760 years
14.3 days
87.2 days
27.8 days
60.0 days

Half-life hours
107500
343
2093
667
1440

If you select a suitable half-life correction then, for example, if a batch is recounted after e.g.
one week, the corrected CPM results are directly comparable with each other since they are
half-life corrected back to the same date and time.
N.B. Always check that the date and time are set to correct values in MicroBeta. The current
time can be seen in the upper right corner of the screen during counting. If these values are
wrong, set them in the System state (see Clock).

72

2.15 Help and Info.

2.15 Help and Info

2.15.1 MicroBeta TriLux Help
The MicroBeta program has a Help function. This means that information can be displayed to
explain what options are open to you at the phase of operation you are in. This information can
be obtained by pressing one of the keys: H, h or ?. In protocol editing press Enter also. These
help keys can be pressed at any stage of the program with one exception. The H keys must not
be used to get help when H has another meaning e.g. when defining active rows in the protocol
editor. Use ? to get help at that point.

2.15.2 MultiCalc Help

At any point in using MultiCalc you can get help by pressing the F10 function key. To get help
about the function of any softkey press the ALT key and the softkey at the same time. Pressing
ALT and F9 gives you a general keyboard help at any part in the program when text mode is
being used for the screen.
To clear a help press ALT and F10.
As experience in operating the instrument grows with time, the need for detailed information
decreases, and the additional information the Help function gives will not need to be accessed
so often.

2.15.3 Selecting info

Info contains a short user manual and information on consumables, normalization and
standardization positions, changes after the previous program version and customizing the
program.
Information can be viewed on the screen or printed. To enter the Info state press I in the Ready
state.
Ready>I
(H)elp
(U)sage
Ch(a)nges

(Q)uit
(C)onsumables
Cu(s)tomizing

(P)ositions
S(y)stem

Info>

73

2.15 Help and Info.

2.15.4 (H)elp
Press H to get help about the various selections.

2.15.5 (U)sage
This is a short user's manual with basics on how to use the instrument.

2.15.6 (C)onsumables
This is an up-to-date list of the currently available consumables.

2.15.7 (P)ositions
This is a list of sample positions in standardization and normalization.

2.15.8 Ch(a)nges
This is a list of additions and changes made to the program after the previous version.

2.15.9 Cu(s)tomizing
The program can be customized using various environment strings. These strings and their
effects are listed under this control letter.

2.15.10 S(y)stem
This is a list of system information and settings.

74

2.16 Interrupt

2.16 Interrupt
2.16.1 Stop
If it is necessary to stop counting quickly, press O (Off). The cassette currently in the counting
position will be moved back to the rack and the program will enter the Ready state.
Note: If you have started counting from MultiCalc and are in MultiCalc mode, you cannot use
the O key. Instead you must press F1 (=STOP), or if the softkeys are not in use, then press any
other key e.g. the space bar to stop counting.

2.16.2 Open the door

If the door is opened during counting, the high voltage is cut off, and the counting stopped. An
error message is displayed on the screen. When the door is closed the program starts counting
the current positions from the beginning.

2.16.3 Next position or assay

If you want to stop the counting of the current samples, press key N (Next position) or E (Next
assay). Counting will be interrupted and the MicroBeta will start to count the next position if N
is pressed or the next assay if E is pressed. If quick view is used and N(Next plate) is pressed,
then MicroBeta starts counting the next plate. Similar keys in MultiCalc mode are F2 (= NEXT
POS) and F3 (= NEXT ASSAY).

2.16.4 Causing a pause in printing

It is possible to cause a pause in printing during counting. Press P (Pause printing). After a
while the computer asks if you want to pause in printing. Answer Y (Yes) to the question to
cause a pause in printing. To resume printing later on press P again. To not pause, answer N
(No). This feature is not included in MultiCalc mode.

2.16.5 Causing a pause in data transfer

A pause in data transfer to the terminal PC can be caused if required. It is recommended that
this feature be used before making a temporary exit from the terminal emulator program to the
MS-DOS of the terminal PC. MicroBeta can measure approximately three cassettes belonging
to the same assay after the pause in data transfer has occurred. If the pause is longer than this,
then more results will not be saved.

75

2.16 Interrupt

To cause a pause in data transfer press D (Disconnect) and answer Y to the question asking
confirmation. Wait until the message Press D to resume data transfer appears on the live
display, then exit from the terminal emulator. After returning to the terminal emulator press D
to resume data transfer.

76

2.17 Loading the cassette rack

2.17 Loading the cassette rack

2.17.1 Cassette rack
The cassette rack is located inside the door and
consists of 16 or 32 shelves, numbered 1 to 16 (32)
from the bottom upwards. The maximum capacity
is therefore 1536 (3072) samples when using 96well plates, and 384 (768) samples when using 24well plates and 6144 (12288) samples when using
384-well plates. Any combination of the two types
of plates accepted by your MicroBeta (96 and 24
or 96 and 384) can be used. The type of cassette
(384, 96 or 24 position cassette) is recognized
automatically by the counter.
Last cassette
position

2.17.2 Loading
The cassette rack is in its middle position when
being loaded. Cassettes should be loaded into the
rack with the sample plate upwards and the ID
labels towards you. The cassette to be counted first
should be inserted on the bottom shelf, numbered
with 1, and the next cassettes on the following
shelves. Empty shelves between cassettes are
allowed, and the first cassette must not necessary
be on shelf no. 1 as long as there is no other
cassette below it.
After loading, close the door properly.

2.17.3 Operation
The movement of the cassette rack, and cassettes
on the transportation system normally occurs
automatically without any need for assistance from
the user. However, manual operation is also
possible. Press O (Operate conveyor) in the Ready
or Count state to get the cassette movement
functions. Press O (Off) to stop the cassette
movement, C (Clear) to clear the conveyor (i.e.

77

First cassette
position

2.17 Loading the cassette rack

move a cassette from the transportation system to the rack and to move the rack to the middle
position), U (Up) and D (Down) to move the rack up and down, and Q (Quit) to return to the
Ready state. (MultiCalc must be in terminal mode if you want to do these operations from
within it).
NOTE: Do not move the rack up or down when a cassette is in the counting position. Always
first clear the conveyor. Re-start the system if the rack and transportation system becomes
blocked.

78

2.18 Luminescence counting

2.18 Luminescence counting

(See chapter 2J.18 for JET information)

2.18.1 Introduction
Luminescence assays come in two main forms; those making use of luminescence that has a
short signal and those making use of that which emits a long stable signal. Both can be
measured with a liquid scintillation counter, however the short emission assays prove difficult
due to the requirements for reagent addition, mixing and temperature control. More recently
assays with a more stable luminescence chemistry have been described and marketed. These
have been targeted towards the molecular biology laboratory and have utilized both traditional
luminometers and photographic film/image analysis to quantify results. With MicroBeta
TriLux, luminescence assays designed to be completed on a solid support e.g. nylon
membrane, glass fibre or coated plate, and in solution can be directly quantified in a shorter
time than with any of the above methods.
Due to the high signal, luminescence reactions can be measured using single PMTs. The
traditional microtitration plates are totally opaque (black or white). These plates are counted
with the upper tube and a 1450-105 cassette is used. The filters are counted with the upper or
lower tube and a 1450-104 filter cassette is used.
A special feature when using MicroBeta as a luminometer is the 1450-466 coloured sealing
tape. The saturation of PMTs is prevented and crosstalk between adjacent positions is
minimized by cutting the intensity of luminescence light with the 1450-466 coloured sealing
tape.
The luminescence units used are LCPS: luminescence counts per second i.e. corrected CPSvalues/100. Before counting the actual samples, the detector and hence the LCPS-values should
be normalized with a luminescence solution as similar to the actual samples as possible.
The normalization is done first and it is made use of when counting the actual samples.

2.18.2 Preparation of normalization samples (96 and 384-well format)

Make a sample solution which corresponds to the samples to be analyzed, i.e. has the same
enzyme and substrate and pipette the same volume as in the samples, into well/membrane G11
(96-well, 1-12 det.), N22 (384-well, 1-6 det.) and M22 (384-well, 12 det.).
An empty background plate is used for background measurement.
The 96-well sample array is as follows, where S1 is the sample solution:
79

2.18 Luminescence counting

A
B
C
D
E
F
G
H

10

11

12

S1

Plate
Close the sample plate with 1450-461 sealing tape or with 1450-466 coloured sealing tape and
place on a 1450-105 cassette for counting.
Filter
Enclose the filter in a 1450-432 plastic sample bag and place it on a 1450-104 cassette; use
1450-466 coloured sealing tape between the sample bag and detector.

2.18.3 Preparation of normalization samples (24-well format)

The procedure is the same as with a 96-position sample support, but the normalization sample
position is D5 for S1.
1
A
B
C
D

S1

2.18.4 Normalization protocol

Edit the normalization protocol so that it is suitable for luminescence measurements. To do this
in the Ready-state press P (P)rotocol then N (N)ormalization protocols.
Select edit (E) and the protocol number.

80

2.18 Luminescence counting

1 Protocol name:
Luminescence
3 Crosstalk correction (Y/N)
N
5 Isotope 1: 1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
9)Luminescence
9
PMT use: 1)Normal 2)Upper
3)Lower
2
10 Counting time [s]
1.0
12 Precision (2 sigma) [%]
0.2
30 Printer output: 1)No
2)Short
3)Long 4)Programmable
3
32 Display output: 1)No
2)Short
3)Long 4)Programmable
3
34 External output: 1)No
2)Short
3)Long 4)Programmable
1
36 File output:
1)No
2)Short
3)Long 4)Programmable
1
40 Change special features (Y/N)
Y
41
Isotope activity setting (Y/N)
N
42
Background sample (Y/N)
Y
Background counting time [s]
60.0
43
Half-life correction (Y/N)
N
45
Use password (Y/N)
N

Edit the protocol parameters. Select luminescence (9) on line 5, isotope 1, and counting time 1s
on (line 11). A background plate is counted only if selected on line 42.

2.18.5 Normalization counting

Start the normalization run in the Ready or Count state using automatic counting and ID codes,
or by selecting Nx, where x is the number of the protocol.

2.18.6 Counting the actual samples

Edit a counting protocol:
In the Ready state select P (P)rotocol then C (C)ounting protocols.
Then select edit (E) and protocol number.
On parameter line 2 select 1 (CPM) and on line 3 select the number of the luminescence
normalization protocol defined above. On line 42 select Y (Background correction) and on the
next line Y (Use normalization background). Edit the other parameters normally.
Start counting in the Ready or Count state using automatic counting and a protocol ID label or
start with the protocol number and press Enter.

81

2.18 Luminescence counting

When counting the samples, the CPS-values are shown on the status display. The stored
normalization factors are used for correcting the LCPS-values in the printout. The normalized
LCPS-values are shown after counting on the status display.

82

2.19 Microtitration plate format output

2.19 Microtitration plate format output

2.19.1 Advantages of plate format
It is often more natural to print out MicroBeta counting results in microtitration plate format
because then it is easier to locate the results for a given sample. Plate format also has the
advantage that it often reduces the paper consumption in printer output.

2.19.2 Plate format selection

Microtitration plate format can be selected by including the keyword PLATE in the
programmable output selection of the counting protocol. The user must then also select the
printout fields of interest. Each printout field (such as CCPM1) will be output as its own table of
6 or 12 columns and 1-8 rows. MicroBeta outputs all the selected tables after the cassette has
been counted. The output can contain both normal and plate format output. Items before the
PLATE word are output in normal format and those after it in plate format.
Note: If you are using MultiCalc Assay protocols you can also make plate maps there. See
section 2.22.4.2 and the MultiCalc manual chapter on Plate operation. Plate mapping is done in
a MultiCalc Assay protocol as part of the CODING section. F6 (PLATE) is the softkey to press.

2.19.3 Restrictions
Some restrictions exist, caused mainly by memory requirements:
-

Plate format cannot be used if the Repeats parameter is greater than 1.

Plate format cannot be used if the spectrum plot (symbol SPECTRA) has been selected for
printer output.

Plate format should be selected for the Display only if results are filed with UltroTerm. In
this case the plate format should come between the data markers #PS and #PE.

A maximum of 4 different printout fields will be accepted resulting in up to 4 tables.

If plate format is selected for more than one output medium (display, printer, external,
result file) then the printout fields listed with the first occurrence of PLATE will be sent to
each medium although output selections would list different fields.

A cassette will be measured totally before the plate tables will be output.

83

2.19 Microtitration plate format output

2.19.4 Examples
Example 1 demonstrates the use of table format. In the output selection PLATE SQP(I) CCPM1
CCPM1% the keyword PLATE selects microtitration plate format. A separate table is created for
SQP(I), CCPM1 and CCPM1% values. If the plate is of the 24-well type and all rows (A,B,C,D)
have been specified, the resulting tables are full of 4x6 results.
Counting protocol no: 33
Name: PlateTEST1
CPM normalization protocol no:40

Fri 28-June-2000 15:11

Assay: - / Prot:33 / Cass: - / Func: - / Cassette No:1 / Shelf: 3 / 4*6

SQP(I)
A
B
C
D

1
373.9
291.6
377.9
290.1

2
348.5
219.1
349.4
222.4

3
382.9
292.2
377.5
290.1

4
352.4
221.5
348.6
220.3

5
379.5
288.5
377.9
289.3

6
348.7
218.1
346.2
218.9

1
89950
86066
92202
86131

2
89374
75547
90716
76157

3
91991
86783
92217
86924

4
91113
76144
91019
75498

5
92526
86622
92012
86034

6
91324
74871
90123
75577

1
1.2
1.2
1.2
1.2

2
1.2
1.3
1.2
1.3

3
1.2
1.2
1.2
1.2

4
1.2
1.3
1.2
1.3

5
1.2
1.2
1.2
1.2

6
1.2
1.3
1.2
1.3

CCPM1
A
B
C
D
CCPM1%
A
B
C
D

Total count rate: 2066932.1 CCPM

End of assay

31.05nCi

84

34.45 kBq

2.19 Microtitration plate format output

Example 2 demonstrates how to include replicate statistics with the microtitration plate format
output. Here the keyword STAT can be used (see also the example in section 2.29.3.3
Statistics/File output). The printout fields defined after STAT will be output only when a
replicate group is ready. With the plate format output this means that the printout defined after
STAT will not be represented in table format but as normal output. In addition, results will be
output at once - so the statistics will appear in output before the plate format raw data. In
Example 2 the output selection: PLATE CCPM1 CCPM2 STAT REPL MEAN1 CV1% MEAN2 CV2%
S1 can be interpreted as:
- PLATE : Use microtitration plate format,
- Collect CCPM1 and CCPM2 values into tables and output those tables when the cassette has been
measured,
- STAT : Do not collect the following printouts into tables but output them when the replicate
mean value is available,
- REPL : Output replicate range (e.g. A01-A03),
- MEAN1 - CV2% : Output these statistical values,
- S1 : Output deviation values (CV1%,CV2%) at the 1-sigma level.
Counting protocol no:33
Name: PlateTEST2
CPM normalization protocol no:40

Fri 28-June-2000

15:22

Assay: - / Prot: 33 / Cass: - / Func: - / Cassette No:1 / Shelf: 3 / 4*6

REPL
A01-A03
A04-A06
B01-B03
B04-B06
C01-C03
C04-C06
D01-D03
D04-D06

MEAN1
29560.1
31609.3
61304.6
68693.1
29861.3
32664.4
62968.0
69314.6

CV1%
15.5
13.5
20.6
16.7
14.4
14.0
20.3
16.4

MEAN2
97524.5
95978.3
87538.6
83309.6
97421.2
96547.0
89112.1
83678.5

CV2%
0.8
0.8
7.9
7.9
0.7
0.7
6.8
8.3

CCPM1
A
B
C
D

1
27775
53926
27296
55602

2
34766
75863
34828
77735

3
26137
54123
27459
55565

4
33792
76060
35104
75730

5
26680
55490
27400
56205

6
34354
74528
35487
76007

2
96759
79595
96844
82171

3
98349
91742
97291
92343

4
96360
80547
96366
79503

5
96441
90815
97342
91738

6
95132
78565
95932
79793

CCPM2
A
B
C
D

1
97465
91277
98128
92820

Total count rate 1:

2:
End of assay

1157926.1 CCPM
2193329.2 CCPM

521.59 nCi
987.99 nCi

85

19.30 kBq
36.56 kBq

2.19 Microtitration plate format output

86

2.20 Micro-volume LSC

2.20 Micro-volume LSC

2.20.1 OptiPhase SuperMix cocktail
In micro-volume counting very small volumes of sample and cocktail have to be mixed.
Traditional cocktails are not well suited for this. A cocktail specifically for micro-volume
counting has been produced, it is called OptiPhase SuperMix. Aqueous samples can be mixed
with a minimum of shaking, and sample loading can be greater than 1 to 1, see the table below.
The values are in ml per 10 ml of OptiPhase SuperMix cocktail at different temperatures.
Solute
Deionized water
0.01M PSB
40% sucrose
1.0M sodium chloride
0.5M sodium chloride
0.1M sodium hydroxide
0.05M Tris HCL
0.1M hydrochloric acid
0.25M ammonium sulphate
0.1M ammonium sulphate
Krebs-Ringer original buffer

15oC
7.50
18.50
13.00
11.00
15.50
17.00
19.50
16.50
14.00
16.00
16.75

20oC
25.00
19.50
10.50
9.50
13.00
17.00
20.00
15.50
13.50
14.50
16.75

30oC
24.50
16.50
10.00
7.50
10.50
17.00
19.00
14.00
11.50
13.00
15.50

The solvent used, di-isopropylnaphthalene (DIN), is in all OptiPhase HiSafe cocktails

including OptiPhase SuperMix. DIN is not aggressive, so it does not attack the majority of
commonly used plastics (e.g. polyethylene, polypropylene, polycarbonate, polyester and PVC)
unlike other LSC solvents. This makes it ideal for use with microtitration plates.

2.20.2 Safety aspects

The toxicological and physico-chemical properties of DIN have been extensively studied (see
the booklet Di-isopropylnaphthalene - a new solvent for LSC, 1989 available on request). The
conclusion of the study was that DIN is classified as a non-hazardous substance within the
meaning of the EEC-Directive of the 18th September 1979 (79/831/EEC). DIN is not subject to
compulsory labelling as laid down in those regulations which are effective in the European
Community. In accordance with UN and EEC International Transportation Regulations, DIN is
not required to be labelled as flammable or hazardous because its flash point exceeds 65oC (it
is in fact 148oC). A biodegradability study of OptiPhase HiSafe II will also be found in the
above mentioned booklet with the conclusion The test compound, OptiPhase HiSafe II was
found to be readily biodegradable by the ISO 7827-1984 (E) method.

87

2.20 Micro-volume LSC

OptiPhase SuperMix, belonging to the OptiPhase HiSafe family, shares these same features
which thus greatly simplifies the requirements involved in its transportation and disposal as
well as showing it to be both safe to use and environmentally friendly. In many countries it can
be disposed of via the drains because of its biodegradability. It is also possible to use a
removable adhesive seal for the microtitration plates so that liquid and solid waste can, if
necessary, be separated and thus disposal costs reduced. However, it is essential to consult with
your Radiological Safety officer and Institute Administrator before entering into any particular
course of action.

2.20.3 Filling wells

Do not overfill wells. In the case of the flexible MicroBeta 96-well plate the maximum total
volume of liquid should not be much above 200 L; rigid plates have a capacity of 350 L.
Splashes of scintillant on the plate should be removed with a tissue prior to applying the
adhesive seal.

2.20.4 Preparation of a quench series

If you are using constantly quenched samples then the preparation of a quench series is not
necessary. However MicroBeta also offers you the possibility to handle variably quenched
samples. In such a case it is necessary to prepare a quench standard series so that the quench
correction necessary to calculate the final DPM values can be made.
The preparation of six samples is adequate for a quench series, although up to 12 samples can
be used with MicroBeta. Each sample should contain the same amount of radioactivity but the
amount of quenching agent in the cocktail should increase incrementally so that the first
sample has no quenching agent and the last the greatest amount. The range of the quench
standards should be such that it covers the expected quench range of the unknown samples and
also that it allows the quench curve to be well defined by the curve fitting program.

2.20.5 96-well plates

Prepare a series of quenched OptiPhase SuperMix samples. E.g. dispense into 5 ml samples of
OptiPhase SuperMix the following amounts of carbon tetrachloride: 0, 10, 30, 50, 75 and 100
L. Mix these samples.
Next dispense the activity to wells A1 to A6, G11 and G12, then dispense 200 L of the
quenched cocktail to each well. Seal the plate and count it as described in chapter 2.11.
The activity used should be e.g. 20 L of tritiated water or some other suitable source giving
about 200 000 DPM per well so that the counting time required to get an acceptable precision
(e.g. 0.5%) can be about 1 min. If you do not mind using a longer counting time you can use
lower activities.

88

2.20 Micro-volume LSC

Note: The total time for a standardization with 200 000 DPM activity would be less than 20
mins whereas for 20 000 DPM it would be 3 hrs to get the same precision.

2.20.6 Chromium release

Specific cytotoxicity can be measured with microvolume samples by counting the supernatant
from the chromium release. Add 25 L of supernatant to 175 L of OptiPhase SuperMix
cocktail. The crosstalk between wells is of the order of 0.06% when using the printed
microtitration plates. Use a counting window setting of 5 - 170.

2.20.7 Microvolume LSC and chromatography

A chromatography eluent gradient with varying ionic strength can be collected in 96-well
plates for evaluation in MicroBeta. The different aliquots will have different levels of
quenching and hence different SQP(I) values. The SQP(I) value is then used to obtain the
efficiency and hence the DPM for the sample.
In order to automate dispensing to a 96-well plate you can use a device such as the Labsystems
Multidrop liquid dispenser.

2.20.8 Scintillation Proximity Assay (SPA)

2.20.8.1 Introduction
The scintillation proximity assay developed by Amersham is an application for which
MicroBeta with the ParaLux counting feature (see section 2.20.9 below) is very well suited.
In SPA, fluomicrospheres coated with receptors or acceptor molecules are used for binding the
ligand to be assayed. These fluomicrospheres also constitute the scintillation medium. Low
energy electrons e.g. tritium beta particles or I-125 Auger electrons, from the radioactively
labelled bound ligand cause light emission from the fluomicrospheres. Labelled, unbound
ligand does not cause light emission because of the dilute concentrations used in bioassays
where the distances between the unbound ligand and the fluomicrospheres is greater than the
maximum range of the low energy electrons; they are absorbed in the aqueous medium.
Consequently there is no need to separate the unbound ligand. The whole assay is performed in
the microtitration well and the result counted in a beta scintillation counter such as MicroBeta.
2.20.8.2 Colour quenching and SPA
If the SPA involves coloured solutions then there may be a need to make a colour quench
correction. Amersham provides colour quench kits for use with MicroBeta. These are:

89

2.20 Micro-volume LSC

H SPA PVT colour quench and calibration kit code TRKQ 7080.

125

I SPA PVT colour quench kit code RPAQ 4030.

(Note: for more details of DPM standardization procedure with MicroBeta see chapter 2.12
DPM Standardization).
Load the standardization plate into a cassette with a STD label in the FUNC position and a
suitable standardization protocol number and load this standardization cassette into MicroBeta.
Edit the standardization protocol and set the standardization parameters shown overleaf. In
particular select the SPA mode (mode 2). This automatically selects ParaLux counting.
Run the standardization. The results will be saved under the standardization number marked on
the cassette.
When you come to run the actual SPA, set the counting protocol for DPM and give the
standardization protocol number selected above. Other parameters should be set according to
the instructions in your SPA kit insert.
2 Sample type: 1)Normal 2)SPA
5 Isotope 1: 1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
ParaLux (Y/N)
ParaLux mode: 1) High efficiency
2) Low background
Discriminator channel
PMT use: 1)Normal 2)Upper 3)Lower
Window 1: (default)
10 Counting time for crosstalk [s]
. . .
20 Number of standards

21 Isotope 1 activity [DPM]

-> 2
-> 1 (H-3) or 2 (I-125)
-> Y
-> 2
-> 150
->
->
-> 600 (sec)
-> 10
-> x (where x is the activity
of a sample in DPM)

. . .
The remaining parameters can be selected to be default.

2.20.9 ParaLux counting

2.20.9.1 What is ParaLux counting?
In the case of scintillating samples e.g. SPA-beads, that are settled on the bottom of a well, the
scintillation light from the sample, has to travel a longer distance through the sample solution
in order to reach the upper photomultiplier tube. The light reaching the lower PMT has to
travel a shorter distance and it does not have to pass through much sample solution. This
90

2.20 Micro-volume LSC

causes a scintillation light detection asymmetry because events may be detected in the lower
tube which are not detected in the upper tube because of quenching. This asymmetry is most
pronounced with heavily colour-quenched samples.
In the normal counting mode, each pair of PMTs works in coincidence, which means that both
must detect the scintillation event for it to be counted. However, in the case described above,
there are true scintillation events from the sample which are detected by the lower PMT but
which are not in coincidence with any event detected by the upper tube. These events would
normally be rejected by the coincidence system.
In ParaLux counting mode, both the coincidence events and the non-coincidence events in the
lower tube are counted. In addition a new quench parameter called AQP(I) (Asymmetric
Quench Parameter) is used. This parameter is derived from the difference between the
coincidence counts and the counts observed in the bottom PM tube. In other words it
establishes a relation between quench and efficiency.
2.20.9.2 Counting modes
ParaLux counting is set in the protocol at the same time as the isotope is selected, see chapter
2.25 Protocols. ParaLux counting has two modes: Low background mode (default) and High
efficiency mode. The difference between them is as follows.
In the Low background mode, the count rate is calculated from just the coincidence events.
In the High efficiency mode the count rate is calculated from the coincidence events plus the
events not in coincidence that are detected in the lower tube. However these latter events are
only registered for pulses that come in channels exceeding the discrimination channel set for
the MCA. You can set this discriminator channel level (the default is 150). Both modes use
AQP(I) as the quench parameter.
Both modes give improved counting performance because of the use of the Asymmetric
quench parameter. The High efficiency mode gives a higher efficiency but also a higher
background than the other mode.

91

2.20 Micro-volume LSC

92

2.21 Monitors and flags

2.21 Monitors and flags

2.21.1 Functions of monitors and flags
Monitors are used to check the quality of samples and flags to notify of e.g. poor sample
quality or that counting has been terminated. They are selected in the programmable output in
protocol editor (see chapter 2.25 Protocols). The flag is included also in the short and long
outputs and the DPM monitor in DPM counting. At the end of this chapter is a list of monitors
and flags with their output headings and possible values.

2.21.2 Statistics monitor

The purpose of this monitor is to warn of unacceptable count rate variation in a sample. The
sample count rate in different periods of the counting time is measured to determine the sample
count rate variation. The variation must be within the range expected from the statistical nature
of radioactive decay. If the variation is outside the range, due to e.g. chemiluminescence, the
sample is recounted. A maximum of two recounts are made before the results from the last
recount are output.
The statistics monitor is not active unless it has been selected in at least one programmable
output.

2.21.3 DPM out-of-range monitor

This monitor evaluates quench levels of samples in DPM counting and compares these with
corresponding quench levels in the standardization. This is done by measuring SQP(I) for a
sample. If it is not within the standardization range the monitor gives a DPM out of range
message.
The DPM out-of-range monitor is included in the short and long outputs in DPM counting. It is
no longer in use if it has been removed from the outputs.

2.21.4 LCPS out-of-range monitor

This monitor evaluates the light intensity of samples in luminescence counting to inform of
possible counter saturation. The monitor is set to HIGH if counter saturation is detected. The
LCPS out-of-range monitor is included in the short and long outputs in luminescence counting.
It cannot be used with the luminescence scaler board.

93

2.21 Monitors and flags

2.21.5 Termination flag

Counting is normally terminated when the specified counting time is reached or when the user
stops it, but in some cases it can be interrupted earlier. Precision, selected on line 12 in the
protocol editor, is such a case (see chapters 2.25 Protocols and 2.6 Counting control). The
termination flag is set to 'PREC' if the precision value has been reached. The counting stops if
the precision has been reached in all detectors.
Another case that terminates counting is maximum counts or 'hot sample'. If the count value in
any channel in any detector exceeds a certain value, usually 50000 counts, the counting stops
and the flag for the particular sample is set to 'MAX'. This prevents MCA channels from
overflowing.
The flag is set to 'INT' if the user interrupts the counting by selecting Off, Next position or
Next protocol.
Item name
Statistics monitor

Heading
STM

DPM out of range monitor

DPMM

LCPS out of range monitor

LM

Termination flag

FLAG

Value
.
REC1
REC2
ERR
.
OUT
.
HIGH
?
.
PREC
MAX
INT

94

Explanation
No statistical variation detected
No variation after one recount
No variation after two recounts
Still variation after two recounts
OK, no out of range detected
Out of DPM range
OK, no out of range detected
Counter saturation detected
Luminescence scaler is used
(saturation cannot be detected)
Normal termination time reached
Precision reached
Hot sample (more than 50 000
counts/channel)
Counting interrupted by user

2.22 MultiCalc operation

2.22 MultiCalc operation

2.22.1 Introduction
MultiCalc is developed from RiaCalc, the famous immunodiagnostic software package with
nearly 10 000 users in hospitals and laboratories all over the world. In contrast to RiaCalc, the
MultiCalc software can be customized to different labelling techniques, it can handle assays in
both sequential and plate format and it supports PC networks. MultiCalc allows result
evaluation, screening, microtitration plate format calculations with free replicate placing, a
wide range of quality control operations, mainframe communication etc.
MultiCalc runs on any IBM PC or compatible. Graphics supported are CGA, EGA, MCGA and
VGA, both colour and b/w.
The contact between MultiCalc and MicroBeta is by means of a special communication
protocol.
MultiCalc is operated by softkeys. It assigns different functions to the ten function keys F1 to
F10 according to the operating level it is on. The starting set of softkey functions is called the
Main menu and is what you get when MultiCalc has been first started up. To get to the main
menu from any other level you have to press the F9 key one or more times plus answer any
questions about exiting and saving that the program may ask you on the way.
What follows here is a description of some of the features which are especially relevant to the
operation of MultiCalc with MicroBeta. The softkey sequences described begin with the main
menu referred to above.

2.22.2 Installation and startup

The installation procedure is as follows.
a) When you install MultiCalc on your PC you will be asked amongst other questions to select
the Technology and the Counter. Technology describes the counting process to be used. In this
case it is beta counting so you must select BETA. Counter is the type of counter MultiCalc is to
be working with. You must select PLATEV3. You will be asked to load a Disk with
communication drivers. Drivers are instrument version specific so please use the instrument
program disk at this point. You can select more than one type of technology and counter at the
same time. Complete the rest of the MultiCalc installation as described in the MultiCalc User
Manual.

95

2.22 MultiCalc operation

If MultiCalc is already installed in the PC but not for MicroBeta then you must go through a
similar procedure to that described above but you select Setup only rather than Installation
from the main installation menu.
(b) Start MultiCalc.
(c) Press F1 (= COUNTER) to get to counter control.
(d) Move the cursor to PLATEV3 in the counter list.
(e) Press F5 (= INSTALL) to install 1450 MicroBeta for MultiCalc. In step a), you were
adding MicroBeta to the list of possible counters to be operated with MultiCalc. Now with this
instruction you are telling which counter you are actually going to work with.
If MultiCalc cannot get MicroBeta into the Ready state it will ask you to press F3 to get into
the terminal mode.
Then press F3 and try to get MicroBeta into the Ready state. If communication does not seem
to work then see chapter 2.31.4.11 Terminal emulators/GenTerm/ Trouble shooting (1) about
how to try to fix the problem. You can also try to press the Esc-key several times.
(f) At the end of installation answer YES to see instructions about MultiCalc/MicroBeta use.
Note: Steps (b) through (f) should be executed when:
- MultiCalc is connected for the first time to MicroBeta
- MicroBeta has been used in another environment (such as with UltroTerm)
- There seems to be some problems
For more details about MultiCalc installation see the MultiCalc User Manual, Installation.

2.22.3 MultiCalc terminal mode

Sometimes you must use terminal mode, e.g. when changing counting, standardization or
normalization parameters. MultiCalc allows you to operate in terminal mode as mentioned in
the previous section. The type of terminal emulation used by MultiCalc is VT52. I.e. when
MultiCalc is in terminal mode MicroBeta sees it as if it was a VT52 type terminal. MultiCalc
must therefore be selected as terminal emulation in the MicroBeta System mode parameter
Ter(m)inal (see chapter 2.30.10 System/Ter(m)inal).
96

2.22 MultiCalc operation

To get to terminal mode select F1 (=COUNTER) from the MultiCalc main menu, followed by
F3 (= TERMINAL). In this mode MicroBeta can be used as with any other terminal emulator.
Press F9 (EXIT) to exit from terminal mode and go back to the normal MultiCalc counter
mode.
Press F10 (HELP) to get assistance in almost every situation.

2.22.4 Assay protocols

The main difference between operating MicroBeta with a terminal emulator or operating it
with MultiCalc in counter mode is in terms of the data handling abilities. In the former case
everything is determined by the MicroBeta counting protocol which is what is described in
most of the other chapters in part 2 of this Instrument Manual. Counting protocols are normal
liquid scintillation counting protocols containing parameters which control counting (see
chapter 2.25 Protocols/Parameters in a counting protocol). These are used (e.g. edited) without
MultiCalc or in the terminal mode of MultiCalc. Assays with only counting protocols can also
be started from MultiCalc, but the results are not evaluated by MultiCalc. The PROT-ID on the
cassettes identifies assays using Counting protocols.
If, however, you are using MultiCalc counter mode then you work with MultiCalc assay
protocols. Assay protocols are connected to counting protocols. An Assay protocol contains
protocol number, name, counting time and precision, plate coding and outputs. Other
parameters, such as isotopes, CPM/DPM mode, corrections, etc., are retrieved from a
corresponding Counting protocol, which is selected in the Assay protocol editor.
Assay protocols can only be edited in MultiCalc (see Editing protocols below). They are stored
in both MultiCalc and in the counter. New assay protocols and updates are transferred to
MicroBeta by using the PARAMETERS function. To get this press F1( = COUNTER)
followed by F2 (= PARAMETERS).
The results are sent to MultiCalc in raw form for handling according to the instructions given
in the MultiCalc assay protocol. The great range of data handling possibilities available with
MultiCalc are described in the User guide to MultiCalc functions and other manuals. See
especially there the chapters on Protocols, Files and Quality control.
Assays using Assay protocols must always be started from MultiCalc. Samples using Assay
protocols are identified by an ASSAY label on the first cassette in a batch. They can have
numbers from 1 to 99.

97

2.22 MultiCalc operation

2.22.5 Using MicroBeta with MultiCalc

2.22.5.1 Editing protocols
MicroBeta controlled counting protocols, i.e. Counting, CPM normalization and DPM
standardization protocols, must be edited in terminal mode. Select COUNTER (F1) in
MultiCalc's main menu (press F9 one or several times to get to this level), move the cursor to
PLATEV3 in counter mode and select TERMINAL (F3). Protocols can then be edited as
described in chapter 2.25 Protocols.
Assay protocols, which are controlled by MultiCalc, must be edited in MultiCalc. Select
PROTOCOLS (F4) in MultiCalc's main menu and a list of Assay protocols is displayed. Select
CREATE (F2) to create a new protocol. Enter name, protocol ID (or number), and select
BETA as the technology and RIA, IRMA or RATIO as assay type.
To edit an existing protocol, move the cursor to the protocol line and press Enter. If the
protocol list is more than one page long the next page can be displayed by pressing the Page
down-key (usually marked PgDn). Similar, the Page up-key (PgUp) is used for scrolling one
page up. Press the Home-key to get the first protocol and the End-key to get the last protocol in
the list. A protocol can also be selected by giving the beginning of its name.
After pressing Enter, the protocol with its parameters is displayed. Use the cursor keys to move
up and down, and press Enter to change the value on a particular line. The alternatives, if any,
are displayed as softkeys (F1 - F8), otherwise the value is given by typing it.
The beginning of the protocol looks as follows:
01 DUAL ASSAY
02 COUNTING TIME, MAX COUNTS
03 MEASURING PARAMETERS

Select dual (press F1) or single (F2) label results handling. Note that dual assay here means the
way that MultiCalc handles the results. This is not the same as the counter's dual label. Instead,
dual label counting is selected by using a dual label counting protocol.
Enter counting time in seconds and, separated by a space, the Max counts or precision value.
Note that this is expressed as 1-sigma percent while the precision in the counter is expressed as
2-sigma percent.
Select the isotope(s) on line 03 MEASURING PARAMETERS using one of the softkeys F1 to
F8. The isotope names can also be typed in. The Counting protocol to be used corresponds to
the selected isotope as shown in the table following:

98

2.22 MultiCalc operation

Key
F1
F2
F3
F4
F5
F6
F7
F8

Isotope
3
H
125
I
14
C
35
S
51
Cr
32
P
3
H/14C
32
P Cerenkov

Counting protocol
91
92
93
97
94
95
96
98

The Counting protocols 91 to 98 are factory made. They may not be deleted and the isotopes
may not be changed. They are connected to factory made CPM normalization protocols with
the same numbering (91-98).
Note: No CPM normalization or DPM standardization is made at the factory, so you must
prepare suitable normalizations or standardizations for the factory made counting protocols.
The Counting protocol number can also be typed instead of giving the isotope. Please check
that this protocol exists and that it has the right isotope(s) and counting mode. If the protocol
does not exist the default protocol (number 0) will be used.
Examples of selections on line 03:
H3
H3/C14
18

- Counting protocol 91 is used.

- Dual label, Counting protocol 96 (3H/14C) is used.
- Counting protocol 18 is used (can be single or dual label)

If you are using a newer version of MultiCalc you will first be asked for the counter instead of
the isotope. Press softkey PLATEV3. A list of counting protocols will be displayed and the
isotopes available will be selectable via softkeys. You can now either select a factory made
counting protocol by pressing an isotope softkey or select a protocol from the list. Scroll the
list by the up and down arrow keys (or PageUp/PageDown for faster scrolling) and make the
selection by pressing Enter.
Note: If you have a newer version of MultiCalc but do not get the counting protocol list, try to
install the counter again. Press F5 (=INSTALL) in the counter mode and the counter will send
the protocol index to MultiCalc.
See also the Introduction to MultiCalc operation manual, the module on Protocol operations.

99

2.22 MultiCalc operation

2.22.5.2 Plate coding

Plate coding is accessed by pressing F1 (CODING) (See the MultiCalc User Manual Plate
operation).
Plate coding requires the replicate number and the sample type to be set. Press F7 to increase or
F8 to decrease the replicate number. Press F1 to F4 to select the sample type. Then press Enter
to get a new line for the next replicate type.
The PLATE-key (F6) allows a sample layout to be defined directly onto the plate. After
pressing F6, a plate is displayed showing the different codes in different wells. Use the cursor
keys to move around the plate. Select EDIT (F1) to edit existing layouts or CREATE (F2) to
define a layout of your own. See the MultiCalc manual for how to make the plate layout.
Press F9 to exit the plate coding.
Again, press F9 to exit from protocol editor. Select F1 to save changes to the protocol or F2 to
exit without saving.
2.22.5.3 Sending assay protocols to the counter
Assay protocols must be stored in both MultiCalc and the counter. After exiting from the editor
the protocol will only be stored in MultiCalc and it must therefore be sent to the counter. Go to
the counter control (press F9 and F1) and select PARAMETERS (F2). Press F2 to send every
Assay protocol or move to the protocol name and press Enter to send a particular protocol.
If you want to check what assay protocols have been sent to the counter, you can use
commands P and M in the MicroBeta Ready state. (This comes from the command list items
(P)rotocols/(M)ultiCalc assay protocol).
There are three functions you can select from:
Show - displays the contents of an assay protocol
Print - prints out the contents of an assay protocol
List - prints the list of assay protocols
2.22.5.4 CPM normalizations and DPM standardizations
Normalizations and standardizations are made in the same way as when using MicroBeta
without MultiCalc (see chapters 2.23 Normalization and 2.12 DPM standardization). Go to the
counter control in MultiCalc and select TERMINAL (F3). Select normalization or

100

2.22 MultiCalc operation

standardization protocol in the Ready state to edit the protocol. Start the counting as normal
from the Ready state.
Normalizations and standardizations can also be started from MultiCalc. Check that a NORM
or STD label is in the FUNC field of the cassette. Select PLATEV3 in the counter control and
press Enter to start counting.
NOTE! The results files from the normalizations and standardizations will not be evaluated by
MultiCalc!
2.22.5.5 Counting unknown samples
Unknown samples can be counted by using MicroBeta controlled protocols (Counting
protocols) or MultiCalc controlled protocols (Assay protocols). The type of protocol is selected
by putting an ID label with protocol number in the PROT field of the cassette for Counting
protocol assays. When using Assay protocols the ID label is attached to the ASSAY field.
The counting of samples using Counting protocols is started as when using MicroBeta without
MultiCalc, i.e. select Automatic counting in the Ready state. It can also be started from
MultiCalc by pressing F1 (=COUNTER) and then selecting PLATEV3 and pressing Enter. The
live data and results are displayed on screen and results are sent to each output medium as
selected in the protocol. Printer outputs are sent to the printer connected to the PC if Print
through terminal is selected in System/Ter(m)inal. Results to file are sent through MultiCalc to
the appropriate disk drive on the PC.
NOTE! The results from assays using counting protocols will not be evaluated by MultiCalc!
The counting of samples using Assay protocols may only be started from MultiCalc. Check
that the protocol number label is in the ASSAY field of the first cassette in a batch. Select
PLATEV3 in the counter control and press Enter to start counting. Live data can be displayed
by pressing F4 (=LIVE). The results of each sample are displayed in plate format if plate
coding has been used. The results are then stored in MultiCalc for further evaluations (see
Stored files and Result files in the MultiCalc manual).
2.22.5.6 Communication protocol
See MultiCalc User Manual Communication protocols.
To change the RS232C port, change only lines from 02 COMM.PORT to 07 COMM.MODE.

101

2.22 MultiCalc operation

2.22.5.7 Communication problems

See chapter 2.31.4.11 Terminal emulation/GenTerm/ Trouble-shooting for information about
how to solve possible communication problems.
2.22.5.8 Printer connection
MultiCalc prints results to the printer connected to the terminal PC with a standard Centronics
cable. In terminal mode MicroBeta controlled assays direct printing also to the same printer.
2.22.5.9 Exit to MS-DOS
In the main menu select first LEVELS (F7) then MS-DOS (F8). See the MultiCalc User
Manual for details of other menu operations.

102

2.23 Normalization

2.23 Normalization
(See chapter 2J.23 for JET information)

2.23.1 What is normalization?

MicroBeta has 1, 2, 3, 6 or 12 detectors allowing it to count the respective number of samples
simultaneously. In order for the results from each detector to be equivalent, irrespective of
small variations in efficiency and background between detectors, it is necessary to determine
the relative efficiency and background of each detector and then correct for it. This is called
normalization.
Normalization is done by measuring an optional background plate followed by one or two
standard samples with defined activity or CPM in each detector. Once CPMs have been
measured the relative efficiencies can be calculated. The efficiency of the detector giving the
highest count rate is then defined to be one and the other detector efficiencies are expressed as
a fraction of this value. These fractions are called efficiency coefficients.
If isotope activity is given, then absolute efficiencies are calculated by dividing the count rates
by the activity. In this way, sample quench and detector absolute efficiencies can be corrected
for.
When measuring a sample with a particular detector, the CPM of the sample is corrected by
dividing the CPM by the efficiency coefficient. In dual label counting, corrections are made
using dual label correction formulae (see 3.3 Calculation methods).
When normalization is done the results are stored with the normalization protocol. The
normalization data can be used by one or several protocols and is selected when editing the
protocol (see 2.25 Protocols).

2.23.2 When is normalization necessary?

Each counting or assay protocol requires normalization data to be stored before it can calculate
corrected CPMs for the samples counted. A fresh normalization must be done when a new
isotope is counted or when counting features, such as isotope or window, are changed. When a
new type of microtitration plate is used, crosstalk normalization may be necessary.

103

2.23 Normalization

2.23.3 Editing normalization parameters

Normalization parameters are stored in normalization protocols. These parameters are shown in
the figure. There can be a maximum of 100 normalization protocols, numbered from 0 to 99.
Protocol number 0 is the default protocol.
Line (1..46, type / to exit, ? for help)

1 ->

1 Protocol name:
->
3 Crosstalk correction (Y/N)
N
4 Number of labels: 1)Single 2)Dual
1
5 Isotope 1: 1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
9)Luminescence
1
ParaLux (Y/N)
N
PMT use: 1)Normal 2)Upper 3)Lower
1
Window 1:
5- 360
10 Counting time for crosstalk [s]
60.0
12 Precision (2 sigma) [%]
0.2
30 Printer output: 1)No
2)Short
3)Long 4)Programmable
3
32 Display output: 1)No
2)Short
3)Long 4)Programmable
3
34 External output: 1)No
2)Short
3)Long 4)Programmable
1
36 File output:
1)No
2)Short
3)Long 4)Programmable
1
40 Change special features (Y/N)
Y
41
Isotope activity setting (Y/N)
Y
Isotope 1 activity (DPM)
200000.0
42
Background sample (Y/N)
N
43
Half-life correction (Y/N)
N
44
Chemiluminescence correction (Y/N)
N
45
Use password (Y/N)
N
46
Special plate: 1)Wallac 2)BP filter
3)Other
1

->
->

->
->
->
->
->
->
->
->
->
->
->
->180000
->
->
->
->
->

Protocols 91 to 98 inclusive are made at the factory for isotopes 3H, 125I, 14C, 51Cr, 32P, dual
label 3H/14C, 35S, and 32P Cerenkov respectively. These may not be deleted because they are
connected to the corresponding counting protocols. Note that no normalization is done for
those protocols. You must use these protocols to do the normalization before they can be used
for counting unknowns.
Press P and N in the Ready state to choose the normalization protocol state. The protocols can
then be edited, copied, deleted, printed and displayed. Press F to get default parameters and E
to edit a protocol. C is used for copying a protocol including normalization data and D for
deleting a protocol.

104

2.23 Normalization

Note: the normalization data will be destroyed when editing an old normalization protocol. The
normalization must therefore be done again for this protocol. Only the protocol name and
password can be edited without destroying normalization data.
See chapter 2.25 Protocols for further details on normalization parameters and editing.

2.23.4 Preparing the normalization samples

Depending on which type of MicroBeta you have it is supplied from the factory with:
(for 1-6 detector instruments with 24 or 96-well capability) a 24-well normalization sample
plate containing standards for 3H and 14C. The 3H standard is in D5 and the 14C standard is in
D6.
(for 12 detector instruments with 96 or 384-well capability) a 96-well normalization sample
plate containing standards for 3H and 14C. The 3H standard is in G10 and the 14C standard is in
G12.
(for other configurations of MicroBeta special arrangements are made).

The sample positions and the activities for the standards are shown on a label on the plate. The
activity values can be inserted into the normalization protocol.

105

2.23 Normalization

Be careful that the samples are in the correct positions when making your own normalization
plates. The positions cannot be changed by modifying protocol parameters. In 24-well plates
the standard for isotope 1 is in D5 and the standard for isotope 2 is in D6. Isotope 1 is the lower
energy isotope in dual label counting. In single label counting the standard is in the isotope 1
position, i.e. D5, with one exception:
the standard for 14C must be in the isotope 2 position in single label counting with 24-well
plates.
This is because the factory made normalization plate could be used for both single label 3H and
14
C normalizations and dual label 3H / 14C normalizations.
For best results, 96-well plates should be used as normalization plates if 96-well sample plates
are to be counted in a 1-6 detector counter. The isotope 1 standard should be in G11 and the
isotope 2 standard in H12.
See 2.7.2 Crosstalk CPM normalization for crosstalk sample positions and 2.18 for
luminescence sample positions.
If background sample is selected on line 42, then thermal backgrounds for count subtraction
are measured automatically. An empty background cassette must be the first normalization
cassette followed by the normalization cassette with standards.
See the table below for sample positions in normalization plates:
Sample position in a normalization plate
24-well
96-well
(1-6 det.)
(1-6 det.)
Isotope 1 standard
Isotope 2 standard
14
C single label

D5
D6
D6

G11
H12

96-well
(12 det.)
Single Dual
G11
G10
G12

384-well
(1-6 det.)

384-well
(12 det.)

N22

M22

2.23.5 Normalization procedure

First place the empty background plate in a cassette and then plate the plate with the
normalization samples in a cassette which is identified with the corresponding normalization
protocol number in the protocol number area and with a NORM label in the function code area.
Insert the cassette into the rack and close the door. Press either A (Automatic counting), or

106

2.23 Normalization

Nxx, where xx is the normalization protocol number, in the Ready or Count state to start
counting. If Nxx is selected the program asks:
Do you want to continue counting after normalization? (Y/N)->

If Y (Yes) is selected, the counting continues after counting the normalization plate, if N (No)
is selected the counting stops after the normalization. If normalization has already been done
for the protocol then the program asks first:
This protocol contains normalization data.
Continue? (Y/N) ->

This is to prevent accidental overwriting of normalization data.

If automatic counting is selected the normalization cassette can be placed anywhere in the rack.
The NORM label is needed in this case.
The counting results for the normalization are output as specified on lines 30 - 37 in the
normalization protocol, and after them the background and detector efficiency values are given.
Note! Normalization and unknown samples should have the same quench level in dual label
counting.

2.23.6 Preset normalizations

There are four preset normalizations available in MicroBeta:
Normalization protocol 1 contains a 3H normalization using ScintiStrip plate 1450-419, dry,
200 L volume with crosstalk correction.
Normalization protocol 2 is the same as 1 but using 125I.
Normalization protocol 3 contains a generally used 3H normalization.
Normalization protocol 4 contains a generally used 14C normalization.

107

2.23 Normalization

108

2.24 P-32 Dot blot quantification

2.24 P-32 Dot blot quantification

Assays utilizing the 32P label and 96-format filtermats, e.g. dot blot hybridizations can be
detected with MicroBeta using the 32P cassette (1450-118). Dried sample membranes can be
counted directly in the cassette, while wet membranes need to be counted in a sample bag, to
avoid any contamination. The counting method does not include any scintillator addition to the
actual filter and thus allows re-probing of the samples. When 32P samples are measured with the
1450-118 cassette the obtained results are quantitative. The 32P counting efficiency is around
70% and background less than 10 CPM. Crosstalk values less than 0.05 % are achieved.
The cassette consists of a cassette body and a lid. In the cassette body there is a steel plate, which
is the same size as the filtermat. The filtermat is aligned on the steel plate according to the edges
and the lid is clicked on the filtermat, so that the small hole is in the upper left corner, the
A1-corner.
To count the 32P cassette in MicroBeta, choose other isotope, upper PMT usage and window
350-1024 in your normalization or counting protocol. No crosstalk correction is needed. If you
use Windows workstation you can use the parameter 32P cassette 8 by 12 for 32P membranes or
you can set up a new parameter with the settings previously mentioned.

109

2.23 Normalization

110

2.25 Protocols

2.25 Protocols
(See chapter 2J.25 for JET information)

2.25.1 Four types of protocols

A protocol is a set of parameters which control the counting of samples. There are four types:
- CPM normalization protocol
- DPM standardization protocol
- MicroBeta Counting protocol
- MultiCalc Assay protocol
A CPM normalization protocol contains parameters used when doing normalization (see
chapter 2.23 Normalization) and a DPM standardization protocol contains parameters for
standardization (see chapter 2.12 DPM Standardization). A MicroBeta counting protocol is
used when counting unknown samples. An assay protocol is the corresponding protocol in
MultiCalc (see MultiCalc). A counting protocol uses either a normalization protocol (in CPM
counting) or a standardization protocol (in DPM counting) to get certain parameters, e.g.
isotopes and window settings. Assay protocols are connected to counting protocols, from
which they get both basic parameters, such as isotopes and counting mode, and special
features, such as background and half-life corrections.
The maximum number of protocols for each type is 100 (0 to 99). Number 0 is the default
protocol, which is used if no protocol number is specified.
Counting protocol number 99 is a so-called GLP (Good Laboratory Practice) protocol. Count
the normalization sample plate supplied with the instrument using this protocol. Do this from
time to time to make sure that the instrument produces consistent results. The background, 3H
and 14C samples are counted with all detectors so background level, CPM values and quench
parameter values can easily be compared. See chapter 3.2 Routine maintenance for more
details.
In this chapter only MicroBeta protocols are described. For further information about assay
protocols see chapter 2.22 MultiCalc and the MultiCalc User manual Protocols.

2.21.2 Listing protocols

To list or edit protocols, press P and then C for counting protocols, N for normalization
protocols or S for standardization protocols in the Ready state. To select the MultiCalc assay
functions as described in chapter 2.22 press M.
111

2.25 Protocols

To display the protocol list, in the Protocols menu press S (Show). You will see a two column
list of protocols containing protocol number, name (if given) and isotopes. The list of the
counting protocols also contains counting mode (CPM or DPM) and the number for the
normalization or standardization protocol used. The lists of the normalization and
standardization protocols have a column containing the date when the normalization or
standardization has been done. An equals (=) character indicates that crosstalk correction is
used.
If there are more protocols than there is space for on the display, press Enter to get the next
page of protocols. When you have found the one you want to look at, type the number of that
protocol and press Enter.
Type L (List) to printout the protocol list and P (Print) followed by protocol number and Enter
to print a particular protocol.

2.25.3 Creating and editing protocols

To create a new protocol with default values, press E (Edit). Choose a protocol number not in
the list and press Enter. The first free protocol number is shown in the menu. Protocols have
already some values in them, either default or old parameters. If you want to set default
parameters, press F (deFault) and select the protocol number.
You will be asked for a line number when starting editing a protocol. Press Enter to start
editing from the beginning of the protocol or type a line number followed by Enter to edit a
certain protocol line. You can always jump between protocol lines by pressing L followed by a
line number and Enter. Type P and press Enter to jump to the previous line. The up and down
arrow keys can also be used.
The value in front of the arrow on each line is the default value. Just press Enter if you do not
want to change this value, or type a new value and Enter if it must be changed. The Ctrl-B (Ctrl
key held down and the B key pressed) and Ctrl-N keys can be used to move within the input
text; the left and right arrow keys can also be used. See 2.1.4 Editing input data.
NOTE: Unnecessary lines in a protocol can be hidden to make the editor shorter. Press Ctrl-K
(Ctrl and K keys pressed at the same time) to hide the current line. To reveal a hidden line, go
to the next line and press Ctrl-U. To unhide all hidden lines go to line selection line in the
beginning of the editor and press Ctrl-A.
Press / and Enter when you are finished with your editing and want to save the changes made.
If you want to exit without saving the new values, press \ and Enter.
112

2.25 Protocols

2.25.4 Example protocol listing

The example alongside shows a typical dual label protocol listing. The parameters are
described in section 2.25.7.
Counting protocol no: 26 Mon 24-May-2000 10:49
Name: Dual CPM
CPM normalization protocol no:
Wallac 1450 MicroBeta V4.4 S/N 4500759
1 Protocol name:
Dual CPM
2 Counting mode:
1)CPM 2)DPM
1
3 CPM normalization protocol
4 Number of labels: 1)Single 2)Dual
2
5 Isotope 1: 1)H-3
2)I-125
3)C-14
4)S-35
5)Cr-51
6)P-32
7)P-32 Cerenkov
8)Other
1
ParaLux (Y/N)
N
PMT use: 1)Normal
2)Upper
3)Lower 1
Window 1:
5-360
6 Isotope 2: 1)H-3
2)I-125
3)C-14
4)S-35
5)Cr-51
6)P-32
7)P-32 Cerenkov
8)Other 3
Window 2:
150-650
10 Counting time [s]
60
12 Precision (2 sigma) [%]
0.2
20 Count all positions in cassette (Y/N)
N
21 Cassette type: 1)96 2)24
1
22
Cassette 1:(96)
Active rows:
AB
Row A: 111111111111
Row B: 111100000000
Row C: 000000000000
Row D: 000000000000
Row E: 000000000000
Row F: 000000000000
Row G: 000000000000
Row H: 000000000000
Cassette 2:(96)
Active rows:
ACEG
23 Plate orientation: 1)Normal
2)Rotated
3)Mirrored 4)Both
1
30 Printer output: 1)No
2)Short
3)Long
4)Programmable
4
31 Printer output:POS CTIME SQP(I) CCPM1 CCPM1%
:CCPM2 CCPM2% PROT SPECTRA
Plot energy scale: 1)Logarithmic [ch]
2)Linear [keV]
3)Bothspectra
1
Plot area [channels]
1-1024
Plot size: 1)Small 2)Medium 3)Large
2
32 Display output: 1)No
2)Short
3)Long
4)Programmable
3
34 External output:1)No
2)Short
3)Long
4)Programmable
1
36 Fileoutput:
1)No
2)Short

113

2.25 Protocols

3)Long

4)Programmable

37 File output: POS CTIME SQP(I) SQP(I)% COUNTS1

:CCPM1 CCPM1% COUNTS2 CCPM2
:CCPM2% FLAG STM PROT
File path:
C:\MB1450\TEST\
File name extension
1
40 Change special features (Y/N)
Y
41 Counting control (Y/N)
Y
Number of repeats
3
Number of replicates
1
Number of cycles
1
42 Background correction (Y/N)
Y
Use normalization background (Y/N)
N
Background sample (Y/N)
Y
Bgnd sample position
G12
Bgnd counting time [s]
60
43 Half-life correction (Y/N)
Y
Half-life 1 [h]
107500.00
Zero time 1 set (Y/N)
Y
Zero date 1
5-Mar-2000
Zero time 1
10:18:46
Half-life 2 [h]
0.00
44 Chemiluminescence correction (Y/N)
N
45 Use password (Y/N)
N
48 Delay between plates [m]
0

2.25.5 Protocol copy and delete

The contents of one protocol can be copied to another by pressing C (Copy). Select the No. of
the protocol to be copied and enter the new protocol number. If you want to delete a protocol,
press D (Delete) and select the protocol number. Press Q (Quit) to return to the Ready state.

2.25.6 Protecting protocols

Protocols can be protected against unauthorized editing by using passwords. The password is
selected on line 45 in the protocol editor. This prevents other users from changing or deleting
your protocol, but they can still use it (e.g. listing, copying and counting with it). The
MicroBeta supervisor has the authority to list the passwords.

2.25.7 Parameters in a counting protocol

1. Protocol name
Type a name for the protocol, maximum 170 characters (only the first 16 characters will be
shown in the protocol list) and press Enter. The name may not start with characters ?, / or \.

114

2.25 Protocols

2. Counting mode
Type 1 for CPM counting or 2 for DPM counting.
3. CPM normalization protocol / DPM standardization protocol
Type the number for the normalization (in CPM counting) or standardization protocol (in DPM
counting) used. Type H or ? to get a list of normalization/standardization protocols. Isotopes
must be entered if no protocol is selected, otherwise the counting protocol uses the same
isotope(s) as in the norm./std. protocol. Type - if no protocol is needed. This means that no
normalization correction is done in CPM counting and no DPMs are calculated in DPM
counting. Lines 4 to 5 are displayed only if no normalization or standardization protocol
number is selected. Information about whether norm./std. has been done is given after line 3.
4. Number of labels
Type 1 for single label counting (one isotope, default) or 2 for dual label counting (two
isotopes).
5. Isotope 1
Select an isotope, in dual labelled samples the isotope with the lower energy. The alternatives
are:
1) 3H (default)

2) 125I

3) 14C

4) 35S

5) 51Cr

6) 32P

7) 32P Cerenkov 8) Other

9) Luminescence
You will be asked for window limits if you select 'Other', otherwise window settings for the
selected isotope are shown.
ParaLux counting can be selected in single label non-crosstalk and non-luminescence
protocols. If ParaLux is selected, then AQP(I) is used instead of SQP(I). ParaLux counting has
two modes: High efficiency and Low background. The discriminator channel that is used to
calculate AQP(I) and High efficiency counts can also be optimized.

115

2.25 Protocols

For PMT use select normal coincidence counting, or use only upper or lower photomultiplier
tubes. Upper is the default if luminescence counting is selected.
6. Isotope 2
Displayed only in dual label counting. The higher energy isotope is selected. The alternatives
are the same as on line 5 with 14C as default isotope.
7. Window 1
Displayed only if 'Other' has been selected as isotope 1. Enter the limits for window 1 in a
range from 1 to 1024.
8. Window 2
Displayed only if 'Other' has been selected as isotope 2. Enter the limits for window 2 in a
range from 1 to 1024.
9. Window 3
Displayed only if 'Other' has been selected as isotope 1 or isotope 2 in dual label counting.
Enter the limits for window 3 in a range from 1 to 1024 or 0 - 0 if no third window is used.
10. Counting time
The counting time is given in seconds. The minimum value is 0.1 and the maximum is 999
999.9 seconds. The default value is 60.0. If quick view is selected then the maximum plate time
is requested. This is the total counting time of the plate without the sample change time. The
default value is 960.0.
12. Precision (2 sigma)
Counting stops when the standard deviation is below the two sigma value in all detectors.
Precision is given in percent in a range from 0 to 99.9. 0 means that no precision limit is used.
The default value is 0.2 %.
20. Count all positions in cassette
Press Y (Yes) if you want to count the whole cassette (default) or N (No) if you want to specify
the positions to be counted. If No has been selected the next parameter will appear:
21. Cassette type
Type 1 if you use cassettes with 96 wells (default) or 2 if the cassettes have 24 or 384 wells.

116

2.25 Protocols

22. Active rows

If you answered No on line 20 you will have to specify the rows to be counted for the first
cassette by typing the letters for these rows. E.g. 'AC' means that samples A1 to A12 and C1 to
C12 will be counted. For a more detailed specification, i.e. giving the positions instead of rows
only, press R (Row coding). In this case specify which positions are to be counted for each
row. The positions are in order from 1 to 12 (or 6 for 24-well plates). Type 1 to count a
position or 0 to skip it. E.g. Row A: ->0011110 means that positions A3, A4, A5 and A6
should be counted. You do not have to enter all the zeros at the end of the line; the row end is
filled with 0 or 1 according to the last typed character. See the following example:
20 Count all positions in cassette (Y/N)
Y
21 Cassette type: 1)96 2)24
1
22
Cassette 1:(96)
Rows:
AB
Row A: 111111111111
Row B: 001111000000
Row C: 000011000000
Row D: 000000000000
Cassette 2:(96)
Rows:
ABCDEFGH
Cassette 3:(96)
Rows:
ABCDEFGH

->N
->
->R
->0011110
->0000110
->0
->AB
->/

An alternative method is to type the column numbers e.g. 5,6,7 equals 000011100000, 1,
equals 100000000000 and 3,, equals 001111111111.
After pressing Enter you will be asked for positions for the next row. The default values are the
same as you have given on the previous row.
Press / when you want to stop specifying counting positions. The rest of the rows will then be
counted as the last edited row. In the example above, the following positions will be counted
for cassette 1: A3, A4, A5, A6, B5 and B6. The following cassettes can then be coded in the
same way as the first one. Press / once again to exit cassette coding. If there are more cassettes
in a batch than there are defined cassettes, the rest of them are counted according to the last
cassette specification.
W can be pressed to code additional cassettes later.
X can be pressed to change the cassette type (96 to 24 (or 96 to 384) and vice versa) for
cassettes 2 to 16, see the example:
23 Plate orientation
Select 2 if rotated plates (A1 at the bottom right corner) are used, 3 if mirrored (A1 at the top
right corner), and 4 if both rotated and mirrored (A1 at the bottom left corner).

117

2.25 Protocols

30. Printer output

There are four printer output alternatives:
1) No
2) Short
3) Long (default)
4) Programmable
1) No
If No is selected no results are printed.
2) Short
If Short is selected, sample results are printed using the short results format: POS, CCPM1,
CCPM1%. In dual label CCPM2 and CCPM2% are added. CCPM is replaced with DPM in DPM
counting. In luminescence counting it is POS, LCPS and LCPS%.
3) Long
The long output contains: PROT, POS, CTIME, SQP(I) (or AQP(I)), CCPM1, CCPM1%, plus
CCPM2, CCPM2% in dual label counting. In DPM counting DPM1 and DPM1% are added. In
luminescence counting the output is PROT, STIME, POS, CTIME, CPS1, LCPS, LCPS%.
4) Programmable
The output items and arithmetic expressions are defined separately on the next line. Sample
quality monitors can also be selected in the programmable output. Press ? or H to get a list of
output items, monitors and rules for using own formulae. See also chapters 2.1 'Beginning
operation', 2.26 'Results', 2.29 'Statistics' and 2.21 'Monitors and flags'.
If SPECTRA is selected, three more lines are shown:
The first asks for the energy scale to be selected. It can be logarithmic with the units in
channels or linear with the units in keV or both of these. Next you are asked to specify the
range of the plot. This will be channels or keV depending on your previous selection. Finally
you are asked for the spectrum size, small, medium or large. A smaller size will give faster
output. The two figures following illustrate the type of plots you may get.

118

2.25 Protocols

Spectrum with logarithmic scale

Spectrum with linear scale

32. Display output
This output is selected when you want to display results on the screen. It can be chosen
independently of the printer output.
Display output has the same output alternatives as the printer output.
The default programmable output, PRINTER means the same data as specified for printer
output.

119

2.25 Protocols

34. External output

This output is selected when sending data to an external device such as a mainframe computer
connected to the MicroBeta via port 2. The data is in ASCII format (text files).
The output alternatives are the same as in printer and display output, except that alternative 1,
No output, is the default value.
The default programmable output, PRINTER means that the same data is specified as for
printer output.
36. File output
The counting results can be stored on a floppy disk or hard disk on the PC or MicroBeta, or on
the server of a local area network (LAN) (section 2.8 Datafiles). The default setting is 1, no file
output. The alternatives are the same as in printer output.
The default programmable output, PRINTER means that the same data is specified as for
printer output.
The result files are named so that the first part of the name (the part before the period) is the
protocol number and the second part (after the period) is a running number indicating the
assay's execution order. E.g. the first file for protocol 5 is named 5.001, the second 5.002, etc.
Result file names for normalizations begin with N (e.g. N1.001) and standardizations with an S
(e.g. S1.001).
The disk drive (A:, B: for floppy disks, C: and D: for hard disks, F: etc. for LAN drives) and
the path (e.g. \MB\DATA\) for the result files are chosen after the File output. This over-rides
any path set in System mode. The drive and path defined in the System state is the default
value (see chapter 2.30.9 System/Data drive).
This is the terminal PC path. If results should be stored on the MicroBeta disk, then the path
must begin with an @, e.g. @B:\DATA\.
If a VT52 or VT100 terminal is used then results are stored on the protocol disk as default
(directory B:\RESULTS), see 2.30.10 Terminal.
The file name extension (the part after the period) can be changed on the next line. This must
be a number in the range 1 to 999.
Note: this line is not available if UltroTerm is used.

120

2.25 Protocols

40. Change special features

Press Y to change special features. These are counting control, background, half-life and
chemiluminescence correction, password and special plate type.
41. Counting control
This and the following questions will be displayed only if Yes has been selected on the
previous line.
Press Y to change the counting control parameters. These are number of repeats, replicates and
cycles.
If N is selected then these parameters have no effect.
The number of repeats is the number of times each sample will be counted. The minimum
value is 1 (default) and the maximum is 99. If repeat is greater than 5, results are not sorted and
statistical results are not printed. Repeats cannot be selected in crosstalk.
The number of replicates specifies how many replicates there are for a sample. This is used for
evaluating sample preparation errors. The minimum value is 1 (default) and the maximum is
99.
The number of cycles is the number of times a batch of sample cassettes will be counted. The
minimum is 1 (default) and the maximum is 99. If the number of cycles is greater that 1 then
you will be asked to give the Cycle delay. This is in minutes and is the time between when one
cycle ends and the next one begins.
See chapter 2.6 Counting control for more information.
42. Background correction
Press Y to use background correction. You can choose between using the background
measured in normalization, giving the position for a background sample or giving a value to be
subtracted from the sample CPM. Answer Yes to the background sample question if you want
to use a background sample. Enter the position for this sample. H12 (96-well, 1-6 detectors),
G12 (12-detectors), D6 (24-well) or P24 (384-well, 1-6 detectors) is the default. Enter also the
counting time (60s is the default). Note that the background sample must be in a position
reached by every detector (see chapter 2.9 Detectors for these positions). The measured
backgrounds are subtracted from CPMs when calculating CCPMs.
Answer No to the background sample question if you want to give a background value. The
value, given in CPM, is subtracted from the sample CPM for an isotope. Two values are given,

121

2.25 Protocols

one for each window, if dual label counting is used. In dual label DPM counting, three values
are given. The value is in the range from 0.0 (default) to 99.9 CPM or 999 999.9 LCPS.
Note: in the normalization protocol press Y if you want to count the background plate.
43. Half-life correction
Press Y if you want to use half-life correction. The half-life for the isotope is displayed, e.g.
107 500 hours for 3H. No half-life value is given for 14C because it is so long. Note that you
must always specify the half-life if you have selected an isotope of type 'Other'. It must be in a
range from 0 (no half-life) to 999 999.99 hours (114 years).
If normalization or standardization protocol is selected on line 3 then half-life is as specified in
this protocol and it cannot be changed in the counting protocol
Press Y on the next line if you want to set zero time, otherwise the activity is corrected to the
time when counting of the first sample of the assay was started.
The zero time consists of date and time. First enter the date in the format DD-MMM-YYYY, where
DD is the day number (1 - 31), MMM the month in letters (JAN, FEB, MAR, APR, MAY, JUN,
JUL, AUG, SEP, OCT, NOV and DEC) and YYYY the year, e.g. 15-JUN-2000. The date can
also be in the format YYYY-MM-DD, e.g. 2000-06-15. Then enter clock time in the format
HH:MM:SS, where HH is hours in the 24 hour mode (0 - 23), MM minutes (0 - 59) and SS seconds
(0 - 59), e.g. 14:05:30. It is not necessary to give seconds. The default value for zero time is
the current time. See also chapter 2.14 Half-life.
44. Chemiluminescence correction
Press Y to use chemiluminescence correction, otherwise N (default). This correction is done
with a correction formula, see chapter 3.3 Calculation methods.
45. Password
Press Y to select a password if you want to prevent other users from changing your protocol.
Enter the password, maximum 12 characters. This will NOT be echoed to the screen. You will
also be asked to retype it to prevent typing errors. The protocol cannot later be edited or deleted
without knowing the password. You must therefore try to remember it. Only the MicroBeta
supervisor has the authority to list the passwords (see chapter 2.30 System/Passwords). He or
she can help you if you have forgotten your password.
46. Special plate
Select Special plate type. Wallac is the standard plate. Betaplate filtermat plates and other 24well plates have different dimensions and are counted using just one detector.

122

2.25 Protocols

However, it is possible to count other 24-well plates with all detectors. This selection comes
after the plate dimension questions. For Betaplate filtermats you can select the 96-well plate
format output. This is a composite of four 24-well plate format outputs. If you select the Other
option for plate type, you must give the plate dimensions. These are the distances of the centres
of the A1, A6 and D1 wells from the plate sides. Special cassettes must be used for Betaplate
filtermat plates and other 24-well plates. The default dimensions 17.3/ 13.4/113.6/71.5 are for
the Costar plate type. For the Falcon plate type the dimensions 14.5/13.4/110.7/71.5 should be
used.
The default dimensions for a special 384-well plate are 11.8/9.0/115.3/76.5. These are the
distances of the centres of the A1, A24 and P1 wells from the plate sides.
Note: this line is not available if 96-well cassette has been selected elsewhere in the protocol.
47. Autoquench correction
Automatic quench correction can be done in single label CPM counting. Isotopes must be 1, 2,
3, 4 or 6, and a non-crosstalk CPM normalization protocol must be selected.
48 Delay between plates
The delay between the end of plate measurement and the start of the next plate measurement
can be specified. The range is 0 to 9999 minutes.

2.25.8 Parameters in a normalization protocol

The parameters are much the same as in the counting protocol with some exceptions.
Information whether normalization/standardization has been done is given after the protocol
name. The following counting protocol parameters are not included in the normalization
protocol:
2.
3.
20.
21.
41

Counting mode
CPM normalization/DPM standardization protocol
Count all positions in cassette
Cassette type
Counting control

The following parameters are new or changed when compared with the ones in the counting
protocol:
3. Crosstalk correction
Press Y to use crosstalk correction otherwise select N. see chapter 2.7 Crosstalk correction for
further information.
123

2.25 Protocols

10. Counting time for background

The counting time for the background and crosstalk counting samples in crosstalk counting is
given in seconds. The minimum value is 0.1 and the maximum is 999 999.9 seconds. The
default value is 60.0.
11. Counting time for standards
The counting time for the standard samples is also given in seconds, from 1 to 999 999.9 with
60.0s as the default value.
41. Isotope activity setting
Press Y to set activities for normalization samples. This is isotope dependent and given in
DPM. The range is from 1000.0 to 999 999.9. The default value is 200 000 for isotope 1 and
100 000 for isotope 2.
If activities are given, absolute normalization is done with efficiencies as count rates divided
by the given activity.
If N is selected, relative normalization is done with efficiencies as a fraction of the highest
detector count rate.
47. Strip plate
Press Y if you use plates where crosstalk is different in the X and Y directions. The crosstalk
sample in G10 is counted in this case.
See chapter 2.23 Normalization for further information.

2.25.9 Parameters in a standardization protocol

The parameters are much the same as in the counting and normalization protocols with some
exceptions. Information whether normalization/standardization has been done is given after the
protocol name. The following counting protocol parameters are not included in the
standardization protocol:
2.
3.
20.
21.
41.
42.

Counting mode
CPM normalization/DPM standardization protocol
Count all positions in cassette
Cassette type
Counting control
Background correction

The following parameters are new or changed when compared with the counting protocol:

124

2.25 Protocols

2. Sample type
Select normal or scintillation proximity assay for sample type. ParaLux counting is set
automatically if SPA is selected.
3. Crosstalk correction
Press Y to use crosstalk correction otherwise select N. See chapter 2.7 Crosstalk correction for
further information.
14. Easy DPM
Press Y to use Easy DPM quench curves. Only the two DPM normalization samples are
counted in standardization.
20. Number of standards
Give the number of DPM standards used for standardization. The value must be in a range
from 2 to 12 (6 in dual label) with 6 as default.
21. Isotope 1 activity
Give the isotope activity in DPM. The range is from 1000.0 to 999 999.9 with 200 000 as the
default value.
23. Isotope 2 activity
Give the Isotope 2 activity in DPM. The range is from 1000.0 to 999 999.9 with 200 000 as the
default value.
24. Standard curve fit selection
Press Y to use standard curve fitting, otherwise N. If No is selected, smoothing spline with
automatic smoothing will be used or linear interpolation if SPA is selected for the sample type.
25. Curve fit method
This line is displayed only if you have selected Yes on the previous line. The methods for
standard curve fitting are:
1) Smoothing spline (default)
2) Interpolation spline
3) Linear interpolation (default for SPA)
4) Linear regression
Select a method by typing the number and Enter. If you have selected the first alternative,
smoothing spline, you will be asked if the method is to include automatic smoothing. Press Y

125

2.25 Protocols

(default) if so. Otherwise press N and you will be asked for a smoothing parameter. Enter a
value between 0.0 (no smoothing) and 1000.0 (maximum smoothing), with 1.0 as default.
26. Edit standard curve
Press Y to edit the quench parameter, efficiency and weight values for a standard curve or to
delete curve points.
41. Adjusted activities
Press Y if you want to adjust the activities for standards, otherwise the activity given on line 21
will be used for every standard. After pressing Y you will be able to change activities for the
same number of DPM standards as given on line 20. You can also change the activities for the
two Easy DPM standards. The range is from 1000.0 to 999 999.9 DPM. The value given on
line 21 is default. See chapter 2.12 DPM Standardization for further information.

126

2.26 Results

2.26 Results
2.26.1 Output items
Different kind of output items can be entered after selecting Programmable output (format 4)
on lines:
30
32
34
36

Printer output
Display output
External output
File output

The possible output items are shown in the tables following:

Note: in these tables y stands for 1 or 2 or 3 and z for 1 or 2
(a) Sample identification data:
AS
POS
SEQ
PR
CA
FU
RP
D
S
CT
SH
CNO
PLATEID
IM
IMI

=
=
=
=
=
=
=
=
=
=
=
=
=
=
=

Value of assay ID for the current plate (0-99)

Sample position on microtitration plate (A01-H12/P24)
Running sample number (1=A01,96=H12,97=A01 on 2nd plate etc.)
Protocol number used (0-99)
Value of cassette ID for the current plate (0-9999)
Value of function code for the current plate (NORM, STD, STOP)
Repeat number of the current measurement
Detector number (1-6)
Spectrum half (A,B).
Cassette type (24, 96, 384)
Shelf number (1-16/32)
Cassette order number (1,2,3,...)
Plate ID
Injector module
Injector module index

(b) Sample counting data:

CTIME
ETIMEz
STIME
DATE
TIME
SQP(I)
SQP(I)%
ISEP
ISEP%
COUNTSy
CPMy
CPMy%
CCPMy
CCPMy%
DCCPMy

=
=
=
=
=
=
=
=
=
=
=
=
=
=
=

Dead time corrected counting time in seconds

Elapsed time in hours for isotope z from zero time z
Counting start time (HH:MM:SS)
Current date (DD-MMM-YYYY)
Current time (HH:MM:SS)
Spectrum quench parameter (0.0-1024.0)
Percentage theoretical error of spectrum quench parameter
Isotope spectrum end point (0.0-1024.0)
Percentage theoretical error of ISEP
Counts for isotope z (or extra window 3)
Count rate (counts per minute) for isotope z (or extra window 3)
Percentage theoretical error of CPMy (i)
Corrected count rate for isotope z (or extra window 3) (ii)
Percentage theoretical error of CCPMy (i)
Absolute theoretical error of CCPMy (i)

127

2.26 Results

DPMz
DPMz%
DDPMz
CLM%
EFFy
STM
DPMM
FLAG
Wy
CPSy
CCPSy

=
=
=
=
=
=
=
=
=
=
=

RESP1
RESP2
RESPz%
LCPS
LCPS%
DLCPS
LM
AQP(I)
AQP(I)%
CPM_C
CPM_NC
CPM_AQP

=
=
=
=
=
=
=
=
=
=
=
=

DPM of isotope z (ii)

Percentage theoretical error of DPMz (i)
Absolute theoretical error of DPMz (i)
Chemiluminescence in window 1
Isotope z (or extra window 3) counting efficiency
Statistics monitor (iii)
DPM out of range monitor (iii)
Termination flag (iii)
Window y limits
Count rate (counts per second) for isotope z or extra window 3
Corr. count rate (counts per sec.) for isotope z
(or extra window 3)(ii)
CCPM1 or LCPS
CCPM2
Percentage theoretical error of RESPz
Luminescence corrected counts per second divided by 100
Percentage theoretical error of LCPS (i)
Absolute theoretical error for LCPS (i)
LCPS out of range monitor (iii)
Asymmetric quench parameter (0.0 - 1024.0)
Percentage theoretical error of asymmetric quench parameter
Coincidence CPM1
Non-coincidence High efficiency CPM1 = CPM_C + CPM_NC
CPM used in AQP(I) calculation

(c) Statistical data:

STAT
S1
REPL
MEANz
CVz%
SEMz%
TCVz%
TSEMz%
SDz
SEMz
TSDz
TSEMz
CHISQz
Pz

= The data specified after this will be output only when a

repeat or replicate mean value is available
= Selects 1 sigma level for all deviation output fields (i)
= Replicate sample range (e.g. A01-A03 if A01, A02 and A03 are
replicates)
= Mean value of CCPMz, DPMz or LCPS
= Percentage coefficient of variation for CCPMz, DPMz or LCPS (i)
= Percentage standard error of mean for CCPMz, DPMz or LCPS (i)
= Percentage theoretical coeff. of variation for
CCPMz, DPMz or LCPS (i)
= Percentage theoretical stand. error of mean for
CCPMz, DPMz or LCPS (i)
= Standard deviation for CCPMz, DPMz or LCPS (i)
= Standard error of mean for CCPMz, DPMz or LCPS (i)
= Theoretical standard deviation for CCPMz, DPMz or LCPS (i)
= Theoretical standard error of mean for CCPMz, DPMz or LCPS (i)
= Reduced chi-square value for CCPMz, DPMz or LCPS
= Probability as a percentage for CCPMz, DPMz or LCPS

(d) Controlling selections:

SPECTRA
NEWLINE
NL
SPSAVE
PROT

=
=
=
=
=

Plot spectra (Printer output only)

Start new line
Start new line (shorthand)
Output spectra in compressed numerical format
Include protocol listing in the beginning of output

128

2.26 Results

PLATE

= Output results in microtitration plate format (see the chapter

Microtitration plate format output)
PRINTER = Output same data as specified for printer (iv)
TXT
= Produces "Start of assay:nn"-message
TAB
= Produces tab-character (ASCII 9) between all output columns to
ease loading of the result file into commercial programs
e.g. spreadsheets
EXCL
= Controls excluding of unrequired parts of the output (v)
FF
= Sends a form feed character to the printer after the
"End of assay" message
NOLIVE = No status display

Notes:
(i) Statistical deviation values will be output normally on the 2-sigma level. However if item
S1 exists in an output selection then for that output device, deviation values will be printed on
the 1-sigma level.
(ii) Corrections made:
(a) Background correction if in protocol either:
42 Background correction = Y (Yes)

and
Use normalization background = Y (Yes)

or
Background sample
= Y (Yes) or
Background subtraction 1 >0.0 (CCPM1 or DPM1) or
Background subtraction 2 >0.0 (CCPM2 or DPM2).

(b) Half-life correction if in protocol:

43 Half-life correction = Yes

and
Half-life 1 > 0
Half-life 2 > 0

(CCPM1 or DPM1) or
(CCPM2 or DPM2).

(c) Chemiluminescence correction if in protocol:

44 Chemiluminescence correction = Yes

(d) Detector efficiency and isotope spillover correction if normalization has been done for the
protocol
e) Quench correction if standardization has been done
f) Crosstalk correction if in normalization or standardization
3 Crosstalk correction = Yes

(iii) Monitors and flags in output (see chapter 2.21 Monitors and flags)

129

2.26 Results

a) Statistics monitor checks count rate variations in a sample and gives the values:
.
REC1
REC2
ERR

No statistic variation detected

No variation after one recount
No variation after two recounts
Still variation after two recounts

b) DPM out-of-range monitor, selectable in DPM counting only, checks quench levels in
samples by comparing with the quench in standardization. The following values can be
displayed:
.
OUT

OK, no out of range detected

DPM out of range

c) LCPS out-of-range monitor, selectable in luminescence counting only, checks counter

saturation. The following values can be displayed:
.
HIGH
?

OK no out of range detected

counter saturation detected
luminescence scaler is used (saturation cannot be detected)

d) Termination flag tells if counting has been stopped. The values for FLAG can be:
.
PREC
MAX
INT

Normal termination
Precision reached
Hot sample (more than 50 000 counts/channel)
Interrupted by user

(iv) Applicable on lines 32 (Display output), 34 (External output) and 36 (File output). Selects
same output as was selected on line 30 (Printer output). However PRINTER does not select
automatically protocol listing (PROT) although PROT exists in printer selection. E.g. to select
protocol listing for file output you must specify PRINTER PROT.
(v) The EXCL output item allows you to customize your output. This can be useful if, for
example, you want to prepare output for a spreadsheet program.
The normal output format of an assay is shown below in the table in the next column.

130

2.26 Results

32-> Data file: F:\USR\GUEST\IHME\33.003

16-> Counting protocol no:33 Thu 27-May-2000 15:38
16-> Name: TEST
16-> CPM normalization protocol no: 40
8->Assay:-/ Prot:33/ Cass:-/ Func:-/
Cassette no: 1/ Shelf:3
/4*6
PlateID:
4-> POS
A01
...
A06

CTIME

CCPM1

CCPM1%

29.7

90680

1.2

29.7

90972

1.2

29.7

86029

1.2

29.7

75349

1.3

75025

1.3

1->
B01
...
B06
...
D06

29.7

2->
8->Assay:-/ Prot:-/ Cass:-/ Func:-/
Cassette no. 2/
4->POS

Shelf:4

CTIME

CCPM1

CCPM1%

29.7

90652

1.2

29.7

90963

1.2

B01
...
B06
...

29.7

86122

1.2

29.7

76239

1.3

D06

29.7

75198

1.3

A01
...
A06

/4*6

1->

16-> Total count rate:4130593.2 CCPM

1859.56 nCi68.83 kBq
32-> End of assay

You will notice that certain lines are tagged with a number and an arrow. In normal operation
these line tags are hidden but they are shown here to help explain this function.

131

2.26 Results

The EXCL-keyword can be used to mask out of the output whichever of the tagged lines you
want left out. To do this you must use the code EXCL followed by two numbers separated by
colons ":". The format is
EXCL:<begin>:<rest>

where:
<begin>

sum of the numbers used to tag lines you want to exclude before and during the
first cassette output (add 64 if not greater than <rest>, see below)
<rest>
sum of the numbers used to tag lines you want to exclude during output from
other cassettes and after the last cassette.
The numbers which can be used (as the example shows) are for <begin>:
32 - datafile path
16 - Protocol name, number and type
8
- ID label contents and shelf number
4
- output column headings
2
- empty line after results from one cassette
1
- empty line after results from one row
For <rest> the numbers are:
32 - end of assay
16 - total count rate
8
- ID label contents and shelf number
4
- output column headings
2
- empty line after results from one cassette
1
- empty line after results from one row
A number resulting from the addition of any combination of these numbers can only be formed
by that combination. So when you give the sum to the counter it knows what combination of
these numbers has gone into making it and hence which lines to omit from the output.
Example: EXCL:72:12 excludes the ID-line before the first cassette (<begin>=64+8) and
excludes the ID-line and headings before the other cassettes (<rest>=8+4). Note that here 64
was added because 8<12.

2.26.2 Changing the format of an output field

The format of an output field (width of field and number of decimals) can be changed. The
syntax is:
<heading>:<width>

132

2.26 Results

or:
<heading>:<width>:<decimals>

where:
<heading> = One of the output items in groups (a), (b) and (c) listed above (except

S1,STAT,REPL).
<width> = The width of the output field in characters (including <decimal>). The program
adds an extra space character between printout fields.
<decimals> = Number of digits to the right from the decimal point (works for real number
values such as CCPM1 only). In the examples following character # denotes a leading space
character and it is supposed that the value of the CCPM1 field is 1234567.123456.
Examples are given in the table at the head of the next column:
FORMAT
OUTPUT
SPECIFICATION
OBTAINED
CCPM1
1234567
CCPM1:13:3
##1234567.123
CCPM1:13:4
#1234567.1235
CCPM1:12:4
1234567.1235
CCPM1:11:4
1234567.123
CCPM1:9:4
1234567.1
CCPM1:8:4
#1234567
CCPM1:7:4
1234567
CCPM1:6:4
1.2E006

EXPLANATION
Default format (=CCPM1:7:0)
13 characters, 3 decimals
Last digit 5 because of rounding
Exactly 12 digits in field
Value too large strip off a decimal
Value too large strip off 3 decimals
Value too large strip off all decimals
Value fits exactly without decimals
Value too large show value in scientific
notation. 1.2E006 means 1.2 times ten
to the power 6.

2.26.3 Entering constant text into an output

Text can be defined between apostrophes (). The syntax is:
<text> or <text>:<width>

where:
<text> = Arbitrary commentary text.
<width> = Width of the text field. If this is greater than the number of characters in <text>

then leading spaces will be produced.

Examples:
SPECIFICATION
"Example"
"Example":5

OUTPUT
Example
Example

"Example":10

###Example

""
"":10

##########

EXPLANATION
Character inside ""s shown.
Format has no effect (format is less than
length of Example).
3 leading spaces output resulting in a total
of 10 characters in a field.
Empty text. No output.
A method to produce 10 spaces.

133

2.26 Results

2.26.4 Defining new output fields

New output fields can be defined. The syntax of a new output is:
[<heading>=<expression>]

or
[<heading>=<expression>]:<width>

or
[<heading>=<expression>]:<width>:<decimals>

where:
<heading> = The heading for the new output field. This must not be included in the headings

listed in paragraph (2.26.1) above. It consists of letters and digits starting with a letter.
<expression> = An arithmetic expression consisting of fixed headings listed in paragraph
(2.26.1)(a) and (b) above (excluding POS, S, STM, DPMM, LM, FLAG, W1, W2 and W3),
numerical constants and operators listed below. Note that statistical values cannot be used in
expressions.
<width> = The width of a printout field in characters (default 12).
<decimals> = Number of digits to the right from the decimal point (default 4).
Operators allowed are:
+

Addition

e.g. CCPM1+100

Subtraction

e.g. CCPM1-100

Multiplication

e.g. 100*CCPM1

Division

e.g. CCPM1/100

Powerfunction

e.g. CCPM1^0.5 = Sqrt(CCPM1)

Abs

Absolute value

e.g. Abs(CCPM1-100)

Ln

Natural logarithm

e.g. Ln(CCPM1)

Exp

Exponent function

e.g. Ln(Exp(CCPM1)= CCPM1

Sqrt

Square root

e.g. Sqrt(4.0)=2.0)

()

Parenthesis

e.g. 10*(CCPM1+100)

Examples:
SPECIFICATION
[CPS1=CPM1/60]

EXPLANATION
Calculates Counts per second and
outputs it in format 9999999.9999.

[RESP=Ln(Abs(CCPM1-102.1087))+0.78]:8:2 An example of a more complicated

formula.The output is in the
format 99999.99.

134

2.26 Results

Note: If a new output field with the heading AQP is given in any programmable output then
this output is used as the quench parameter instead of SQP(I) or AQP(I).

2.26.5 Short output mode

If you select option 2 (short output mode) on protocol lines 30, 32, 34 or 36 then the program
produces as output:
POS CCPM1 CCPM1% (CCPM2 CCPM2%)

Results for each isotope will have their own line.

The corresponding output for DPM counting is:
POS DPM1 DPM1% (DPM2 DPM2%)

For luminescence counting the output is:

POS LCPS LCPS%

2.26.6 Long output mode

If you select option 3 (long output mode) on protocol lines 30, 32, 34 or 36 then the program
produces as output:
PROT POS CTIME SQP(I)(or AQP(I))CPM1 CCPM1 CCPM1% (CCPM2, CCPM2%)

Results for each isotope will have their own line.

The corresponding output in DPM counting is:
PROT POS CTIME SQP(I) (or AQP(I)) CPM1 CCPM1 CCPM1% (CCPM2 CCPM2%) DPM1
DPM1% (DPM2 DPM2%)

For luminescence counting the output is:

PROT STIME POS CTIME CPS1 LCPS LCPS%

135

2.26 Results

2.26.7 Default formats of output fields

The default format of printout fields is shown in the list here:
Note: Fields are always separated by one space that is not included in the field length.
Note: in these tables y stands for 1 or 2 or 3 and z for 1 or 2
(a) Sample identification data
AS:2
CA:4
CNO:3
CT:3
D:2
FU:4
PLATEID:12
POS:3
PR:2
RP:2
S:1
SEQ:4
SH:2

xx
xxxx
xxx
xxx
xx
xxxx
xxxxxxxxxxxx
xxx
xx
xx
x
xxxx
xx

(b) Sample counting data

AQP(I):6:1
AQP(I)%:7:1
CCPMy:7:0
CCPMy%:6:1
CCPSy:9:1
CLM%:6:1
COUNTSy:7
CPMy:7:0
CPMy%:6:1
CPM_AQP:7:0
CPM_C:7:0
CPM_NC:7:0
CPSy:9:1
CTIME:8:1
DATE:11
DCCPMy:9:1
DDPMz:9:1
DLCPS:9:1
DPMz:7:0
DPMz%:6:1
DPMM:4
EFFy:6:4
ETIMEz:9:3
FLAG:4
ISEP:6:1
ISEP%:5:1

xxxx.x
xxxxx.x
xxxxxxx
xxxx.x
xxxxxxx.x
xxxx.x
xxxxxxx
xxxxxxx
xxxx.x
xxxxxxx
xxxxxxx
xxxxxxx
xxxxxxx.x
xxxxxx.x
xx-xxx-xxxx
xxxxxxx.x
xxxxxxx.x
xxxxxxx.x
xxxxxxx
xxxx.x
xxxx
x.xxxx
xxxxx.xxx
xxxx
xxxx.x
xxx.x

136

2.26 Results

LCPS:9:1
LCPS%:6:1
LM:4
RESPz:9:1
RESPz%:6:1
SQP(I):6:1
SQP(I)%:7:1
STIME:8
STM:4
TIME:8
Wy:13:1

xxxxxxx.x
xxxx.x
xxxx
xxxxxxx.x
xxxx.x
xxxx.x
xxxxx.x
xx:xx:xx
xxxx
xx:xx:xx
xxxx.x-xxxx.x

(c) Statistical data:

CHISQz:6:1
CVz%:6:1
MEANz:9:1
Pz:6:1
REPL:7
SDz:8:1
SEMz:8:1
SEMz%:6:1
TCVz%:6:1
TSDz:8:1
TSEMz%:6:1
TSEMz:8:1

xxxx.x
xxxx.x
xxxxxxx.x
xxxx.x
xxx-xxx
xxxxxx.x
xxxxxx.x
xxxx.x
xxxx.x
xxxxxx.x
xxxx.x
xxxxxx.x

137

2.26 Results

138

2.27 Robotic loading interface

2.27 Robotic loading interface

(See chapter 2J.27 for JET information)
The robotic loading interface is a special door/adapter system that brings a cassette outside so
that a robotic arm can change the plate.
In the MicroBeta program the robotic loading interface is selected as follows:
(S)ystem
(L)evel
(T)esting
Detector/shelf (u)sage
Use shelves: 1)1 2)16 3)32 ->1
(L)evel
(N)ormal counting.

The operation is controlled using the RS-232C port 2 (external port) of MicroBeta. When the
cassette is outside, MicroBeta sends the text Ready and carriage return / line feed. The possible
commands are:
H or ?
Nxx
Dxx
xx
A

- list commands,
- start normalization using protocol number xx,
- start standardization using prot. number xx,
- start counting using protocol number xx,
- start counting using IDs.

All Ready commands must be followed by a carriage return / line feed or #. Counting can be
interrupted with O. After a plate has been measured and the cassette is outside, MicroBeta
sends the text Continue and carriage return / line feed. At this point O stops counting and any
other character continues counting.
If plates are counted in the inverted position (A1 at the bottom right corner) then Plate
orientation: Rotated should be selected in the counting protocol. This selection reorders results
so that outputs POS and SEQ are correct.

139

2.27 Robotic loading interface

140

2.28 Safety and radioactive materials

2.28 Safety and radioactive materials

The following comments about precautions and safety measures in handling radioactive
materials are included as a guide and are not intended to be fully comprehensive. More
complete details may be found elsewhere, for example in the booklet SAFE HANDLING OF
RADIONUCLIDES, published by the International Atomic Energy Agency, Vienna; this may
be recommended as a useful code of practice appropriate to radio-chemical laboratories.
Unless a specially designed radio-isotope laboratory is used, limitations should be placed on
the amount of active material in the laboratory area depending on toxicity and type of chemical
operation. For high toxicity material and wet chemical operations involving the risk of spillage,
the IAEA recommend a maximum activity of about 10 Ci.
Personnel should be properly trained in the safe handling of these materials, maximum levels
of stored activities should be set, proper records should be kept, and a definite monitoring
schedule maintained.
The areas where samples are handled should be kept clean and free of dust. This is most easily
accomplished if all surfaces are as smooth as possible and if the minimum number of extra
surfaces is introduced into the room. Lastly it is extremely important to store all radioactive
materials in a separate room to which access is restricted.

141

2.28 Safety and radioactive materials

142

2.29 Statistics

2.29 Statistics
2.29.1 When statistical output is available
MicroBeta software can produce statistical output values in one of the following situations:
(a) Value of Repeats-parameter (protocol line 41a) is greater than 1 and less than or equal to 5.
If Repeats is greater than 5 or if 384-well plates are used then statistical output is not produced.
(b) Value of Replicates-parameter (protocol line 41b) is greater than 1.
(c) Both (a) and (b) hold.
If the user does not specify statistical output values and format then default statistical output
format will be used. In this format a table will be output after the last repeat or after the
replicate group. The columns of the table are shown below (the symbol used in programmable
output is shown in parenthesis):
In this table z stands for 1 or 2
MEAN
=
THEOR.ERROR(%)=
OBS.ERROR(%) =
CHI-SQUARED
=
PROBABILITY(%)=

Mean value of CCPMz,DPMz or LCPS (MEANz)

Theoretical standard error of mean as percentages (TSEMz%)
Standard error of mean as a percentage (SEMz%)
Reduced Chi-squared for mean (CHISQz).
Probability as a percentage (Pz).

See the chapter 3.3 Calculation methods for exact definitions.

The first row of the statistics table shows values for CCPM1, DPM1 or LCPS and the second
row (only in the case of a double label assay) shows values for CCPM2 or DPM2. See the
printer output of the example in 2.29.3. The default for the above values will be output at the 2sigma level. If the user uses Programmable output selection and includes symbol S1 in one of
the programmable output selections then values will be output at the 1-sigma level.

2.29.2 Restrictions
If the user selects programmable output and includes any of the following statistical values
(except S1 and STAT):
MEAN1, CV1%, SEM1%, TCV1%, TSEM1%, CHISQ1, P1, SD1, SEM1, TSD1, TSEM1
MEAN2, CV2%, SEM2%, TCV2%, TSEM2%, CHISQ2, P2, SD2, SEM2, TSD2, TSEM2 REPL

143

2.29 Statistics

then a default statistical table summary will not be produced. Instead you must specify in the
programmable output selection:
- which statistical values to output,
- how to locate these values in the output and
- which kind of formats are to be used to output the values.
Statistical values cannot be used in arithmetic expressions. The default values will be output at
the 2-sigma level unless symbol S1 exists in the selection. The external output of the example
in 2.29.3 demonstrates a compact statistical output where only values MEAN1, SEM1%, MEAN2
and SEM2% have been selected. They have been output on the same line as the last replicate in
order to reduce paper consumption.
The file output of the example in 2.29.3 demonstrates the use of the control symbol STAT. The
output values defined after STAT will be output only when the last replicate or repeat result has
been output. This can be used e.g. to output the statistical data on its own line, as in the
example following, or to produce replicate mean values when using Microtitration plate
format. See the next page for the examples of statistical output.

2.29.3 Examples of statistical output

The following protocol (only relevant parts shown) demonstrates the different statistical output
formats:
30 Printer output:
32
34
35
36
37

40
41

1)No
2)Short
3)Long
4)Programmable
3->
Display output:
1)No
2)Short
3)Long
4)Programmable
3->
External output: 1)No
2)Short
3)Long
4)Programmable
4->
External output: POS CTIME CCPM1 CCPM1% CCPM2 CCPM2% MEAN1 SEM1% MEAN2
:SEM2%
->
File output:
1)No
2)Short
3)Long
4)Programmable
4->
File output: POS CTIME CCPM1 CCPM1% "":6 CCPM2 CCPM2% STAT NL "MEAN":12
:MEAN1 TSEM1% SEM1% MEAN2 TSEM2% SEM2%
->
File path:
C:\MB\TEST\->
File name extension
1->
Change special features (Y/N)
Y->
Counting control (Y/N)
Y->
Number of repeats
1->
Number of replicates
3->
Number of cycles
1->

144

2.29 Statistics

Because symbol S1 does not exist in any of the programmable (lines 35 and 37 above) output
selections the deviation values are represented at the 2-sigma level. The output defined by the
protocol above for each output medium looks like the following:
POS
A01

CTIME
9.9

SQP(I)
388.3

A02

9.9

383.2

A03

9.9

375.5

CPM
29737
112834
29997
110761
32362
111652

CCPM
29737
112834
29997
110761
32362
111652

CCPM%
2.9
1.5
2.8
1.5
2.7
1.5

FLAG
.
.
.
.
.
.

3 replicates:
A01-A03
MEAN
CCPM1 30699.44
CCPM2 111749.09

THEOR.ERROR(%)
1.62
0.85

OBS.ERROR(%)
5.44
1.07

CHI-SQUARED
11.25
1.60

PROBABILITY(%)
0.00
20.23

2.29.3.1 Printer output

Printer output selection (line 30 above) demonstrates the default statistics output format. In
single label counting only a line for CCPM1 is generated.
2.29.3.2 External output
External output selection (lines 34 and 35 above) demonstrates a very compact statistical
output format. The program did not use the default statistical output format because there were
statistical output fields in the programmable output i.e. MEAN1, SEM1%, MEAN2 and SEM2%.
Here the user was not interested in all of the statistical values which can be selected.
POS
A01
A02
A03

CTIME
9.9
9.9
9.9

CCPM1
29737
29997
32362

CCPM1% CCPM2
2.9 112834
2.8 110761
2.7 111652

CCPM2%
1.5
1.5
1.5

MEAN1

SEM1%

30699.4

5.4

MEAN2 SEM2%
111749.1

1.1

2.29.3.3 File output

File output selection (lines 36 and 37 above) demonstrates the use of the control switch STAT.
The output items to the left of STAT (POS CTIME CCPM1 CCPM1% :6 CCPM2 CCPM2%) are
output for each individual result. The empty text definition :6 is used to shift CCPM2 and
CCPM2% seven (=6+1) characters to the right to leave space for the second line SEM1%. The
output items to the right of STAT (NL MEAN:12 MEAN1 TSEM1% SEM1% MEAN2 TSEM2%
SEM2%) are output only when the replicate (or repeat) mean value is available. Then at the first
NL a new line is output. MEAN:12 outputs a commentary word MEAN under the CTIME of the last
replicate. MEAN1 (MEAN2) outputs a mean value under the CCPM1 (CCPM2) of the last replicate
etc.

145

2.29 Statistics

POS
A01
A02
A03

CTIME
9.9
9.9
9.9
MEAN

CCPM1
MEAN1
29737
29997
32362
30699.4

CCPM1%
TSEM1%
2.9
2.8
2.7
1.6

CCPM2
MEAN2
112834
110761
111652
5.4 111749.1

SEM1%

146

CCPM2%
TSEM2%
1.5
1.5
1.5
0.8

SEM2%

1.1

2.30 System

2.30 System
(See chapter 2J.30 for JET information)

2.30.1 Selecting System

System settings control the general way MicroBeta works - its defaults, hardware connections
etc. To enter the System state press S in the Ready state. You are then given a number of
options as example 1 shows. These are described in more detail here.
Example 1
(H)elp
(C)ount

(I)nfo
(P)rotocols

(S)ystem

Ready>S
(H)elp
(E)rrorbeep
(C)lock
(D)ata drive
Customi(z)ing

(Q)uit
(S)tatus display
(R)S-232C
Ter(m)inal

MS-D(O)S
(V)ersion
(P)rinter
P(a)sswords

System>

2.30.2 (H)elp
Press H to get help about the various selections.

2.30.3 (E)rror beep

User or instrument error normally causes a beep. This can be switched off by selecting N.
Example 2
System>E
Error beep (Y/N) Y ->N

2.30.4 (S)tatus display

Normally when counting is occurring its status is shown on the Terminal PC screen (see
chapter 2.5 Counting for more details). The counting status display can be switched off or only
currently counted positions can be shown.
Example 3
System>S
Status display:

1)No
2)Current positions
3)Whole plate

147

3->

2.30 System

2.30.5 (V)ersion
Show the program and MS-DOS version numbers.
Example 4
System>V
MicroBeta program V4.4. Copyright (C)
Wallac Oy, 1997. All rights reserved.
MS-DOS V3.2. Copyright (C) Microsoft
Corporation, 1987. All rights reserved.

2.30.6 (C)lock
This allows the date and time to be set. MicroBeta has a calendar clock with battery back-up so
you do not have to set it each time the power has been switched off, normally only when the
instrument is installed. Details of setting the clock are given in chapter 2.3 Clock. When you
set the clock the Terminal PC date and time are also set if GenTerm is used.
Example 5
System>C
Date 12-Jul-2000 -> 13-Jul-2000
Time
14:44 -> 9:30
Set clock(Y/N)->Y

2.30.7 (R)S-232C
There are three RS-232C communication ports which can be used:
1 for the Terminal PC
2 for an external computer
3 for the printer
You can specify the communication parameters for each of these ports separately. Give the
number of the port and press Enter.
The first parameter that can be changed is the speed of transferring the data, the Baud rate. The
possibilities are: 300, 600, 1200, 2400, 4800 or 9600.
The default value for the terminal and external ports is 9600. For the printout port it is 4800. If
these values are not suitable then select the ones you need.
Next is the parity selection, you can have None, Odd or Even where None is the default.

148

2.30 System

The number of data bits can be 8 or 7 where 8 is the default.

The number of stop bits can be 2 or 1 where 2 is the default.
Lastly you can select the kind of checking process for the data transfer, the handshaking. It can
be None, DTR or XON/XOFF where the default is DTR.
Confirm the selection of these parameters by typing Y twice. When you have done this the
program prints the original and the new settings and returns to the System state if serial port 2
or 3 setting is in question. However when changing the terminal port (port 1) setting the
program then shows a help text that informs you to set your terminal to the same settings which
were selected for MicroBeta before confirming the new setting. When ready press any key. The
program should then ask:
Do you see this text (Y/N)?

If the text appears it is quite sure that the communication works and you can press Y to confirm
the change of terminal port setting. If the text does not appear it is obvious that your new
terminal and MicroBeta serial port settings differ. In that case check from the printer the new
MicroBeta setting and modify your terminal setting if required until you see the above text. If
you cannot make the communication work you have still one possibility. Set the terminal to the
original serial port setting. Switch off the counter and switch on again. Restarting MicroBeta at
this phase returns the original terminal port communication setting. Note: You can lose
communication with the counter also by changing the serial port setting of your terminal alone.
Usually then some strange characters appear around the screen and commands do not function.
To get communication functioning again you can change terminal settings in the following way
if you do not remember the terminal port setting of MicroBeta:
- Try different baud rates. Press e.g. space bar after setting each baud rate.
- If there is no success then change the number of data bits (usually 7 or 8 bits come into
question) and try different baud rates again.
- Usually parity and handshake differences should not prevent normal manual terminal use.
- Another method is to switch the counter off and on again.
The correct parameters (terminal type, baud rate and data bits) are set automatically after
instrument restart. If they differ from the current parameters then a text like the following is
shown: Incorrect terminal baud rate. 9600 bps used. Please set terminal RS232C parameters in (S)ystem /(R)S-232C.

149

2.30 System

When it seems that communication is working, go to the MicroBeta System-mode and check
the serial port 1 settings and make sure that they are identical with settings of the terminal.
Example 6
System>R
RS-232C port number
1->
Baud rate: 1)300 2)600 3)1200
4)2400 5)4800 6)9600
6->
Parity: 1)None 2)Odd 3)Even
1->
Data bits
8->
Stop bits
2->
Handshake: 1)None 2)DTR 3)XON/XOFF 2->
Set terminal port parameters
(Y/N)->Y
Are you sure?
(Y/N)->Y
Do you see this text (Y/N)?Y

2.30.8 (P)rinter
Set the printer type and the number of characters per inch. The options are: IBM or Epson FX
and the number of characters per inch 10, 12 or 17. The defaults are Epson FX and 12 cpi.
Example 7
System>P
Printer type:

1)IBM

2)EpsonFX

Printer charac.per inch:

1)10

2->

2)12

3)17

2->

2.30.9 (D)ata drive

Select diskette or hard disk drive of the terminal PC or MicroBeta on which the results are to
be stored (when using UltroTerm define the result file path in UltroTerm Result filing options).
The result file path can also be given in each protocol.
Example 8
System>D
Data drive:
Path name:

1)A:

2)B:

3)C:

4)Path

4->

C:\MB\RESULTS\->

The options are:

1 = diskette drive A. Select this if you have:
(a) a single drive terminal PC (no other possibilities)
or

150

2.30 System

(b) a hard disk PC but you want to store results on the diskette (e.g. to be transferred
somewhere else).
2 = diskette drive B. Select this if you have a dual diskette drive terminal PC. Then terminal
emulator software can be kept in drive A all the time (e.g. to enable automatic restart after
power failure).
3 = hard disk drive C. Select this if your terminal PC is equipped with hard disk and you want
large storage capacity. If the GenTerm terminal emulator is used, then results will be stored
into the directory which was active when GenTerm was started.
4 = full disk drive and directory path. Use this if options 1-3 do not match your requirements.
E.g. if you want to store results:
(a) on a hard disk drive other than C: (e.g. D:),
(b) on a local area network (LAN) connected to the terminal PC,
(c) in a specific directory or
(d) on the MicroBeta drive.
If 4 is selected then the program asks for the full drive and directory path. Here understanding
of MS-DOS drive and directory concepts is needed. The directory path given here will be used
as such without any checking and modifications in front of the file name. Note that the
directories must exist before any data can be sent to them. Use the command MKDIR in DOS
to make new directories.
Examples (referring to examples a-d above):
(a) Path name: D: ->
- Text D: will be inserted in front of result file name. E.g. D:2.003 , the file will be stored in the
active directory on drive D.
(b) Path name: F: ->
- Here it is supposed that drive F: belongs to the LAN server. Text F: will be inserted in front
of the result file name. E.g. F:17.002 , the file will be stored in the active directory of the LAN
server drive F.
(c) Path name: C:\MB\RESULTS\ ->
- Text C:\MB\RESULTS\ will be inserted in front of result file name. Note that if you specify an
exact directory path for the result files then the path name must end with backslash \. For
example the path C:\MB\RESULTS\3.023 stores result file 3.023 into the directory
\MB\RESULTS\ of hard disk drive C.

151

2.30 System

(d) Path name: @B:\DATA\->

- Results are stored on the MicroBeta protocol disk B in directory DATA. The character @ at the
head of the path shows that the MicroBeta disk is to be used for storing the results.

2.30.10 Ter(m)inal
Select the terminal emulation (see chapter 2.31 Terminal emulators for details) that your
terminal or terminal PC uses. The additional questions depend on the emulation selected. The
possible emulations are:
1 = GenTerm in WT emulation mode. This emulation is fully tailored to be used with
MicroBeta, so the only things the user has to specify are (see example 9):
Example 9
System>M
Terminal emulation:
1)GenTerm 2)UltroTerm 3)MultiCalc
4)VT52
5)VT100
6)MBW
2->1
Number of lines on screen
24->
Print through terminal (Y/N)
N->
Do you accept the new terminal setting? (Y/N)->N

- the number of lines on the screen.

- whether the printer should be connected to the terminal PC or not (see 2.30.10.5).
2 = UltroTerm (see example 10). Select this if you are using the UltroTerm terminal emulator.
UltroTerm V2.0 or later is required.
Example 10
System>M
Terminal emulation:
1)GenTerm
2)UltroTerm
3)MultiCalc
4)VT52
5)VT100
6)MBW
2->
Number of lines on screen
23->
Print through terminal (Y/N)
N->
Do you accept the new terminal setting? (Y/N)->Y
Prepare your terminal ready for emulation change.
Press any key when ready.

3= MultiCalc VT52. Extensions to enable result file saving to PC disk when using MultiCalc.

152

2.30 System

4 = VT52 (see example 11). Select this if you are using a terminal emulator that can provide
only this emulation.
5 = VT100. This emulation is quite popular in commercial terminal emulation programs. If
your terminal emulator implements both VT52 and VT100, select VT100 because it supports a
better range of MicroBeta features i.e. counting results displaying (screen attributes, result
window scrolling etc.).
6 = MBW (MicroBeta Windows workstation VT52). If this is selected then results sorting and
live data automatic sending must be specified. The workstation program sets the terminal and
other settings automatically.
Additional questions:
2.30.10.1 Number of lines on screen
Possible values from 15 to 25, the default value is 24 lines. Some terminal emulator programs
can scroll the whole screen if something is written at the lower right corner of screen (e.g.
column 80 on line 24) which mixes the status display. If you encounter this problem give here
a smaller value (e.g. 23 lines). You can select a smaller value also if your terminal PC cannot
display 24 lines (e.g. some portable PCs can display 16 lines).
2.30.10.2 Block graphics
For VT52 and VT100, default Y=Yes. If your terminal shows some strange characters around
the status display and it seems that it cannot display block graphic characters, select N=No.
2.30.10.3 Send result files to terminal
For VT52 and VT100. The method of sending result files to the terminal PC depends on the
terminal emulator software. Often a terminal emulator can capture data between a start string
and a stop string. If your terminal emulator is capable of doing this then answer Y=Yes. Then
MicroBeta stores a result file at first on the protocol diskette and sends it to the terminal as one
block as:
Start of assay:XX
...
... Result data
...
End of assay

where xx is the protocol number.

If your terminal emulator cannot capture data then answer N=No. Then the result files are
stored on the protocol diskette of MicroBeta but not transferred to the terminal PC.

153

2.30 System

Note 1: Result file will be output if protocol line 36 FILE OUTPUT is set to other than 1 (No
output).
Note 2: See chapter Results for details.
2.30.10.4 Automatic result file deletion
For VT52 and VT100, default Y=Yes. The capacity of the protocol diskette is limited (300KB
- 1.3MB depending on the number of protocols) and it is capable of storing the results from at
least 20 cassettes if the output selection for FILE OUTPUT does not specify too many different
printout fields. If this selection is set to Y=Yes then the result files on protocol diskette will be
deleted automatically when starting counting and after sending a result file to the terminal PC
is complete. If you want to ensure that your data has been transferred to another diskette or to
some PC then answer N=No. In that case after starting counting the program confirms deletion
before operation by asking:
Delete result files saved on protocol disk?

and you can answer N=No to reject deletion or Y=Yes to accept deletion.
2.30.10.5 Print through terminal
Select here where to connect the printer. Answer:
N=No if you do not need to connect the printer to the terminal PC. Connect instead the printer
to serial port 3 of MicroBeta with serial cable 1221-502.
Y=Yes to connect the printer to the terminal PC. In this case connect the printer to the parallel
port of the terminal PC with a Centronics cable 1221-122.
2.30.10.6 Exit terminal setting
To exit terminal setting press slash / and Enter on any line. Otherwise, after you have answered
all the questions described above, the program confirms that you accept the change. If not,
press N, or else press Y which causes the following message to appear:
Prepare your terminal ready for emulation change. Press any key when ready.

Now you can make the preparations possibly required for the emulation change (e.g. to switch
from GenTerm to UltroTerm). When ready press any key. Then if the change succeeded the
program returns to the System state. However if the message Terminal type check failed!
appears (see example 11) it means that MicroBeta has found that the terminal program does not
use the same emulation as was selected (MicroBeta sends an identity inquiry to the terminal
and the answer sent by the terminal emulator software is different for each emulation -

154

2.30 System

GenTerm, UltroTerm, VT52, VT100). Answer N if you want to cancel the new terminal
emulation. However if you know that your terminal has the correct emulation or you want to
switch to the new emulation later you can answer Y.
Example 11
System>M
Terminal emulation:
1)GenTerm 2)UltroTerm 3)MultiCalc
4)VT52
5)VT100
6)MBW
2->4
Number of lines on screen
24->
Block graphics (Y/N)
Y->
Send results file to terminal (Y/N)
N->
Automatic result file deletion (Y/N)
Y->
Print through terminal (Y/N)
N->
Do you accept new terminal setting? (Y/N)
->Y
Prepare your terminal for emulation change.
Press any key when ready
Terminal type check failed!
Do you accept the new terminal setting? (Y/N)->N

2.30.11 P(a)sswords
Press A to print protocol passwords, entered on line 45 in the protocol editor (see chapter 2.25
Protocols). This is possible only if no System password is in use or in Setup mode (see
2.30.15).
Example 12
System>A
Printing protocol passwords, press / to exit

2.30.12 Customi(z)ing
Press Z to set environment strings to customize the program. The environment string name and
value must be given. For a list of possible strings, see (I)nfo/Customi(z)ing. Giving D as the
value sets the default value for the string.
Example 13
System>Z
Customized: MENU = N
Environment string name
MENU
Environment string name

-> MENU
-> D
-> /

155

2.30 System

2.30.13 MS-D(O)S
Press O to get to the MicroBeta MS-DOS. You must confirm this twice as example 14 shows,
after which the text MicroBeta A:\> appears:
Example 14
System>O
Exit to MS-DOS? (Y/N) ->Y
Are you sure? (Y/N) ->Y
Type MB to start the program again
MicroBeta A:\>MB
(H)elp
(I)nfo
(C)ount
(P)rotocols

(S)ystem

Ready>

Type MB and press Enter to get back to the MicroBeta program.

Note: Avoid going to MS-DOS in routine work. The main reasons to go there are to make a
backup copy of the protocol or program diskette.

2.30.14 (Q)uit
Press Q to quit from the System state.

2.30.15 Se(t)up mode

If a system password is used then the system mode menu is quite short (see example 15).
Example 15
(H)elp
(Q)uit
MS-D(O)S
(E)rror beep (S)tatus display (V)ersion
(C)lock
Se(t)up mode
System>T
Password->
(H)elp
(Q)uit
(E)rror beep (S)tatus display
(C)lock
(R)S-232C
(D)ata drive Ter(m)inal
Customi(i)zing

MS-D(O)S
(V)ersion
(P)rinter
P(a)sswords

System>

156

2.30 System

All the functions that can affect routine use are hidden. The person responsible for the
MicroBeta system can make the hidden functions visible if he or she knows the password.
Press T to get into the setup mode. The program then asks for the password. Type the password
and press Enter. The characters you type are not displayed on the screen. If you enter a wrong
password the program displays the message:
Incorrect password

Otherwise the program shows the full System menu.

To set a system password, exit from the MicroBeta program to MS-DOS as described earlier.
After the text MicroBeta A:\> appears, type PASSWORD xxxx and press Enter (xxxx in the
above means the new password). If you forget the password then set a new one.

157

2.30 System

158

2.31 Terminal emulators

2.31 Terminal emulators

2.31.1 Introduction
MicroBeta can be used with a variety of different terminals and terminal emulator programs.
Most of the chapters in part 2 of this manual describe terminal operations. The only exception
is when the MultiCalc counter program is used. This chapter describes operation with two
general purpose terminal emulator programs from Wallac - GenTerm and UltroTerm, and
VT52 and VT100 terminal emulators (see below for an explanation of the terminology). For
information about MultiCalc see chapter 2.22 MultiCalc. You can select whichever of these
terminal emulations is most suitable for your needs.

2.31.2 Terminology
2.31.2.1 Terminal
This consists of a keyboard, display and electronics, and is not usable as a general purpose
computer.
2.31.2.2 Terminal PC
This is any general purpose personal computer that can run a terminal emulator program.
2.31.2.3 Terminal emulator program
This is a program which allows a PC to be used as a terminal and often implements extra
features such as data logging, multiple terminal emulations etc.
2.31.2.4 Terminal emulation
This is a control language that can be used to command the terminal to perform simple
operations such as clear display, move cursor, use underline etc. The control language consists
usually of plain ASCII text, control codes such as backspace and escape sequences (character
sequences started by the Esc-character).
Terminal manufacturers have introduced several different terminal emulation languages e.g.
VT52, VT100 and Wallac WT.
2.31.2.5 MS-DOS
MS-DOS means Microsoft Disk Operating System (a product of Microsoft Corporation). MSDOS enables the user to operate on disks, files, directories, execute programs etc. The
MicroBeta program itself and usually the terminal emulator program run under MS-DOS.
When you exit from the MicroBeta program to MS-DOS which is running in the instrument
microcomputer you see the prompt text MicroBeta A:\>. In contrast when you exit from the

159

2.31 Terminal emulators

terminal emulator program to the MS-DOS of the terminal PC the prompt usually shows either
the current disk drive (e.g. A:>) or the current directory (e.g. C:\GENTERM>). For details about
using MS-DOS see the MS-DOS User's Manual of your terminal PC.
2.31.2.6 Temporary exit from a terminal emulator program
Temporary exit from a program means that the current program remains in the memory but you
can run MS-DOS commands and other (small) programs in the remaining free memory. To
return to the original program type EXIT and press Enter.
2.31.2.7 Exit from a terminal emulator program
Exit from a program means that the current program is terminated totally; to return to it you
must start the program again.
2.31.2.8 Backup
It is recommended that you make a backup copy of your terminal emulator program diskette(s)
before installation. Use the MS-DOS command DISKCOPY at the MS-DOS prompt of the
terminal PC. See the MS-DOS User's Manual of your terminal PC for details.
2.31.2.9 Start up of the terminal PC
A terminal PC can be started up using one of the following methods:
- switch on power
- if power is on, press the Ctrl, Alt and Del keys simultaneously or press the reset button (this
button does not exist in all PC models).

2.31.3 Conventions
Manual references
Interpret notations such as System/Ter(m)inal/Block graphics as:
- chapter = System
- section = Ter(m)inal
- topic = Block graphics

160

2.31 Terminal emulators

2.31.4 GenTerm
2.31.4.1 Product number and manual
1221-243 GenTerm terminal emulator
1221-921 GenTerm User Manual
2.31.4.2 Version
V2.B or later
2.31.4.3 Installation
(a) For details see the GenTerm User Manual three last pages.
(b) When the installation program asks:
Select emulation mode:
1. Normal VT-52 terminal
2. Optimized Wallac Terminal (WT)

then answer 2 (WT emulation).

(c) If installation was made on hard disk then select the way how GenTerm should be started as
follows (these instructions suppose that GenTerm was installed into directory C:\GENTERM):
(1) If GenTerm should start automatically after terminal PC start up, copy lines of
C:\GENTERM\ GTERM.BAT to the file C:\AUTOEXEC.BAT e.g. using the COPY command:
COPY C:\AUTOEXEC.BAT+C:\GENTERM\GTERM.BAT C:\AUTOEXEC.BAT

(2) Execute GenTerm always from the directory C:\GENTERM.

(3) Insert the directory C:\GENTERM to the MS-DOS PATH variable.
(4) Copy file C:\GENTERM\GTERM.BAT into one of the directories listed in the MS-DOS PATH
variable.
d) In the GenTerm setup menu set the Buffer length to a minimum of 100 characters
2.31.4.4 Starting GenTerm
(a) If installation was made on diskette either:
insert terminal diskette into drive A: and start up the PC
161

2.31 Terminal emulators

or
type GENTERM at the MS-DOS prompt and press Enter.
(b) If installation was made on hard disk then you have three possibilities to start GenTerm
depending on the type of installation:
- Start up the terminal PC (installation (c)(1))
- Type CD C:\GENTERM and press Enter then type GTERM and press Enter (installation (c)(2))
- Type GTERM and press Enter (installation (c)(3) and (4))
2.31.4.5 Changing parameters in GenTerm
The main reason for communication problems is that the counter is configured to use one
emulation and the terminal emulator uses some other emulation (see 2.31.4.11 Troubleshooting
below). If there still seems to be communication problems after changing to the same
emulation in both the counter and the PC the reason can be that there are different
communication parameters in the counter and the emulator. Here is a quick guide for how to
change the communication parameters in GenTerm. For further information see the GenTerm
User Manual.
1. Press the Ctrl and Break keys to get to the GenTerm main menu (or in VT-52 mode press
F8).
2. Use the down arrow key to go to Set terminal parameters and press Enter.
3. Select an item from the Set-Up menu by pressing the up or down arrow keys.
4. Change the value of the selected item by pressing the left or right arrow keys.
5. Store the setup on disk as default settings by pressing F3.
6. Press F9 to return to the terminal emulator mode.
Section 2.30 System/(R)S-232C in this manual describes how to change communication
parameters in the counter. This section also contains the default parameter settings.
2.31.4.6 Special features
GenTerm WT emulation emulates the IBM ROM BIOS keyboard and display functions. If
communication problems arise see the Troubleshooting section below.

162

2.31 Terminal emulators

2.31.4.7 Restrictions
Do not change GenTerm to ADM-3A or VT52 emulation.
2.31.4.8 Result files
The destination disk drive and directory can be set in System/(D)ata drive or in Protocol/File
output.
The MicroBeta program uses a special GenTerm WT emulation to direct output to the display,
the printer and the result file. Result file saving is automatic and it is enough for you to only
select on the Protocols/File output-line which data should be saved.
2.31.4.9 Printer connection
The MicroBeta program can automatically direct printer output through GenTerm to the printer
connected to the terminal PC with a standard Centronics cable. The printer is normally
connected to port 3 on MicroBeta.
2.31.4.10 Exit to MS-DOS
To get to the menu press Ctrl-Break or Ctrl-Scroll Lock if the Break-key is missing. Select
either Exit to DOS temporarily (temporary exit) or Exit from GenTerm to DOS (total exit),
press Enter and follow the instructions. During counting it is recommended that you type D
(see chapter 2.16 Interrupt) before temporary exit from GenTerm to MS-DOS. It is then also
possible to exit totally from GenTerm and to use the terminal PC for running other programs.
After returning to GenTerm press D again to allow MicroBeta to send data to GenTerm.
See the GenTerm User Manual for details of other menu operations.
2.31.4.11 Troubleshooting
(1) Communication problems:
Communication problems (e.g. characters are displayed correctly but the keyboard does not
function) will occur if MicroBeta is configured to use GenTerm WT emulation and the
terminal emulator uses some other emulation (UltroTerm,VT52,...) or vice versa.
If you suspect that kind of situation then restart -MicroBeta by putting power off and then on
again. Terminal type, baud rate and data stop bits are set automatically after instrument restart
(see 4.7.2 and 4.7.3). Another method is (if the instrument is in the Ready state) to press
several times the key that sends the Escape character to MicroBeta (usually labelled Esc; in
UltroTerm F3).
MicroBeta should respond: Please check terminal type in (S)ystem/Ter(m)inal!.

163

2.31 Terminal emulators

After doing this the keyboard will work and you can then set terminal emulation in
System/Ter(m)inal. If the message above does not appear then the problem is probably in the
serial communication (cable, RS-232C protocol).
(2) Printer errors:
GenTerm shows an error window. Solve the printer problem (power off, off-line mode, paper
out etc.) and then press R (Retry) to continue printing. If you cannot fix the problem press A to
abort printing and then P until the MicroBeta program asks Pause printing (Y/N). Then answer
Y.
(3) Diskette and hard disk problems:
When the diskette or hard disk becomes full, GenTerm shows a message window that enables
you to:
(a) Change diskette: insert a new formatted diskette (the result file continues on the new
diskette with the same name)
(b) Give a new file name: e.g. direct the result file to another disk drive
(c) Stop logging data
Select the recovery method using the left and right cursor keys, press Enter and follow the
instructions.
If the result diskette is missing or is unformatted etc., then GenTerm displays a Drive not ready
window. Put a properly formatted diskette into the correct drive and press R to continue result
saving. If you cannot solve the problem then press A to stop saving data.

164

2.31 Terminal emulators

2.31.5 UltroTerm
2.31.5.1 Product number and manual
1221-244 UltroTerm 2 Terminal Emulator
1221-922 UltroTerm 2 User Manual
2.31.5.2 Version
V2.0 or later
2.31.5.3 Installation
(a) For details see the UltroTerm 2 User Manual chapter 3 Installation.
(b) Floppy disk installation: follow steps 1-8 as described in the manual. When you reach step
9, instead type INSTALLF 1205 and press Enter.
Note: MicroBeta uses the same mode as Wallac 1205 Betaplate. The number 1205 is therefore
also used with MicroBeta. The number can be changed in communication parameter setting.
(c) Hard disk installation: follow steps 1-2 and 4-7 as described in the manual, in step 3 instead
type INSTALLH 1205 and press Enter.
(d) If installation was made on hard disk then select the way in which UltroTerm should be
started (supposing UltroTerm was installed into directory C:\UTERM):
(1) If UltroTerm should start automatically after the terminal PC start up, then copy lines of
C:\UTERM\ START.BAT to the file C:\AUTOEXEC.BAT e.g. using the COPY command:
COPY C:\AUTOEXEC.BAT+ C:\UTERM\START.BAT C:\AUTOEXEC.BAT

(2) Execute UltroTerm always from directory C:\UTERM.

(3) Insert the directory C:\UTERM into the MS-DOS PATH variable.
(4) Copy the files START.BAT, TASK1.BAT and BPNAME.EXE from the directory C:\UTERM
into one of the directories listed in the MS-DOS PATH variable.
Check that COMMAND.COM exists in the root directory (C:\) of the terminal PC hard disk.

165

2.31 Terminal emulators

2.31.5.4 Starting UltroTerm

(a) If installation was made on diskette either
- insert the terminal diskette into drive A: and start up the PC
or
- type START at the MS-DOS prompt and press Enter.
(b) If installation was made on hard disk then you have three possibilities to start UltroTerm
depending on the type of installation:
- Start up the terminal PC (installation (d)(1))
- Type CD C:\UTERM and press Enter then type START and press Enter (installation (d)(2))
- Type START and press Enter (installation (d)(3) and (4)).
2.31.5.5 Changing parameters in UltroTerm
The main reason for communication problems is that the counter is configured to use one
emulation and the terminal emulator uses some other emulation (see 2.31.5.11 Troubleshooting
below). If there still seems to be communication problems after changing to the same
emulation in both the counter and the PC, the reason can be that there are different
communication parameters in the counter and the emulator. Here is a quick guide for how to
change the communication parameters in UltroTerm. For further information see the
UltroTerm 2 User Manual.
1. Press the Esc key to get to the UltroTerm main menu.
2. Use the down arrow key to go to Set RS232 comm's protocol and press Enter.
3. Enter communication port number and press Enter.
4. Select an item from the Set-Up menu by pressing the up or down arrow keys.
5. Change the value of the selected item by typing the new value or by selecting from the list
using the Home and End keys.
6. Confirm the change by pressing Enter.
7. Store the setup on disk as default settings by answering Y(es) to the last question.

166

2.31 Terminal emulators

8. Press Esc twice to return to the terminal emulator mode.

Section 2.30 System/(R)S-232C in this manual describes how to change communication
parameters in the counter. This section also contains the default parameter settings.
2.31.5.6 Special features
- UltroTerm uses an extended VT52 terminal emulation.
- UltroTerm allows file transfer to a computer, built-in file editing and printing.
2.31.5.7 Restrictions
- UltroTerm can serve only one instrument when in 1205 mode.
In 1205 mode MicroBeta controls printing, so the F7 key does not work.
2.31.5.8 Result files
Select Auto filing start string and Auto filing stop string in Filing parameters. A column
selection marker, e.g. # can be used with programmable output, see 2.26.3. See the UltroTerm
User Manual for details.
2.31.5.9 Printer connection
The MicroBeta program can automatically direct printer output through UltroTerm to the
printer connected to the terminal PC with a standard Centronics cable. The printer is normally
connected to port 3 on MicroBeta.
2.31.5.10 Exit to MS-DOS
To get to the menu press the Esc-key. Select either Terminal-DOS gateway (temporary exit) or
Exit the program (total exit), press Enter and follow the instructions. When the menu is on the
screen press the Esc-key to get back to the terminal mode. During counting it is recommended
that the user types D (see chapter 2.16 Interrupt) before temporary exit from UltroTerm to MSDOS. It is then also possible to exit totally from UltroTerm and to use the terminal PC for
running other programs. After returning to UltroTerm press D again to allow MicroBeta to
send data to UltroTerm. See UltroTerm 2 User Manual/Menu for details of other menu
operations.
2.31.5.11 Troubleshooting
(1) Communication problems
See GenTerm/ Troubleshooting (1)

167

2.31 Terminal emulators

(2) Printer errors

UltroTerm shows an error window. Solve the printer problem (power off, off-line mode, paper
out etc.) and then press the number 1 key to continue printing.
(3) Diskette and hard disk problems
See the UltroTerm User Manual chapter 4.

168

2.31 Terminal emulators

2.31.6 VT52 and VT100 compatible terminals and terminal emulators

2.31.6.1 Installation
This depends on the product, see the manuals which accompany the product.
For cabling see the chapter 4.1 Installation.
Define the terminal features in System/Ter(m)inal (see below for details and chapter 2.30
System/Terminal).
2.31.6.2 Special features
The terminal understands standard VT52/VT100 escape and other control codes.
2.31.6.3 Restrictions
If the product does not understand VT52/VT100 block graphics characters then set
System/Ter(m)inal/Block graphics to N=No.
2.31.6.4 Result files
There are several possibilities:
(1) MicroBeta automatically saves results defined on the Protocol/36 File output line into
directory B:\RESULTS on the protocol diskette. You can copy files from there if your PC can
read standard MS-DOS, 3.5 inch, 1.44MB diskettes.
(2) If you set System/Ter(m)inal/Send result files to terminal to Y=Yes then
MicroBeta sends the result file saved as in (1) at the end of each assay as one block of text in
the format:
Start of assay:xx
...
...
End of assay

where xx is the protocol number. The terminal emulator can now capture the data within Start
of assay and End of assay.
(3) It is possible to define the following environment strings in the MicroBeta AUTOEXEC.BAT.
SET
SET
SET
SET

SOPN
SCLS
SBEG
SEND

=
=
=
=

<open>
<close>
<begin>
<end>

(open result file, name follows)

(close result file)
(save characters until <end> is found)
(end character saving)

169

2.31 Terminal emulators

In the above, <open>, <close>, <begin> and <end> can be any character sequences which do
not include the VT52/VT100 control codes and do not appear in the outputs. For the terminal
emulator the data coming from the RS-232C port looks as follows:
...
<open><path> xx.yyy <CR>
...
<begin> POS CTIME CCPM1
<end>...
...
<begin> A01
60 102356
<end>...
...
<begin> A02
60 115294
<end>...
...
...
<close>

(Display/printer output)
(Open file xx.yyy in directory <path>)
(save this data into file xx.yyy)
(save this data into file xx.yyy)
(save this data into file xx.yyy)

(close file xx.yyy)

where xx=protocol number, yyy=running result file number, <CR>= carriage return character
and <path> = drive and directory set in System/(D)ata drive.
2.31.6.5 Printer connection
VT52 and VT100 terminal emulations have control codes to direct output to the printer
connected to the terminal. These codes are:
- VT52:
<esc>W
<esc>X

(Printer controller mode on)

(Printer controller mode off)

- VT100
<esc>[5i
<esc>[4i

(Printer controller mode on)

(Printer controller mode off)

All characters between printer controller mode on and off codes will be transferred to the
printer. All other characters will be displayed. However usually some control character
(NULL, backspace etc.) not directed to the printer can disable graphics printing.
2.31.6.6 Troubleshooting
For communication problems see GenTerm/Troubleshooting (1).

170

2.31 Terminal emulators

2.31.7 UTMAC
2.31.7.1 Product number and manual
1221-170 UTMAC 1.1 Terminal Emulator for Macintosh.
1221-926 UTMAC 1.1 User Manual.
2.31.7.2 Version
V.1.1 or later.
2.31.7.3 Installation
(a) for details see UTMAC User Manual chapter 3 Installation
(b) If you want to run UTMAC automatically on starting up the Macintosh, make an alias of
the appropriate UTMAC setup document and put it into the Startup folder.
2.31.7.4 Starting UTMAC
To start UTMAC double-click on the icon of the appropriate setup document or on the icon of
the UTMAC application itself. At the present time there is no UTMAC emulation mode in the
counter, but UTMAC is 100% compatible with the UltroTerm emulation mode.
2.31.7.5 Changing parameters in UTMAC
The main reason for communication problems is that the counter is configured to use one
emulation and the terminal emulator uses some other emulation, so check that the counter is set
to UltroTerm terminal emulation mode.
If there still seems to be communication problems after changing to the correct emulation
mode, the reason can be that there are different communication parameters in the counter and
the emulator. Here is a quick guide for how to change the communication parameters in
UltroTerm. For further information see the UTMAC User Manual.
1. Choose Communication from the Settings menu of UTMAC.
2. Modify the appropriate parameters: baud rate, number of data bits, number of stop bits,
parity and type of handshake.
3. Press the OK button to confirm the changes.
4. Save the modified parameters in the appropriate setup file using the Save or Save As
command from the File menu.
Section 2.30 System/RS-232C in this manual describes how to change communication
parameters in the counter. This section also contains the default parameter settings.

171

2.31 Terminal emulators

2.31.7.6 Special features

UTMAC uses an extended VT52 terminal emulation.
UTMAC allows file transfer to Macintosh and printing.
2.31.7.7 Result files
Select the automatic filing start and stop strings in the Filing dialogue. A column selection
marker (data marker) can be used with programmable output, see 2.26.3. See the UTMAC User
Manual for details.
Result files will be saved in the same directory where the current UTMAC setup document lies.
2.31.7.8 Printer connection
The MicroBeta program can automatically direct printer output through UTMAC. However, it
is not printed immediately but gathered into a temporary printer output file. You can print the
output by pressing the Print button that appears at the top of the terminal window or by
selecting Print Printer Output from the File menu. After printing, the contents of the temporary
file will be deleted. It is recommended that you print the printer output only when the counter
is in the Ready state.
2.31.7.9 Exit to Finder
If you are running System 7 or MultiFinder you can always switch to other programs and let
UTMAC work in the background. If you want to stop counting temporarily, type D (see
chapter 2.16 Interrupt) before switching to some other application. Press D again when back in
UTMAC to resume counting.
When you want to exit completely from UTMAC, choose Quit from the file menu. If there is
some printer output pending, you will be asked whether or not you would like to print it. If you
have modified the setup parameters you will be asked to save them.
2.31.7.10 Troubleshooting
(1) Communication problems
See GenTerm/Troubleshooting (1)
(2) Disk full
UTMAC periodically checks the free space left on volume(s) used for filing and warns you if
there is too little space. If any of the volumes gets really full UTMAC halts processing of data
sent by counter and displays a warning message. You should switch to Finder and make some
space or alternatively you can close the data files. Processing of counting data continues
automatically when there is enough free space again or when there are no data files open. Note

172

2.31 Terminal emulators

however, that you cannot close the temporary printer output file which is always located in the
System folder of the boot volume. So if you use the print through terminal option of
MicroBeta, make sure that you have enough free space for printer output on the boot volume.

173

2.31 Terminal emulators

174

2.32 Thermostat option

2.32 Thermostat option

When the Thermostat option is installed it allows the temperature of the upper detectors to be
reduced and stabilized. This lowers the luminescence counting background because the cooling
decreases the photomultiplier tube thermal background. E.g. changing the detector temperature
from 27o C to 20 o C decreases the thermal background by about 60 %.
The thermostat option can be used to cool the upper detectors up to 9 o C below room
temperature.
The detector temperature can be selected in the System-state by using the menu selection
'(-)Detector temp'.
System settings with the Thermostat option
(H)elp
(C)ount

(I)nfo
(P)rotocols

(S)ystem

Ready>S
(H)elp
(E)rror beep
(C)lock
(D)ata drive
Customi(z)ing

(Q)uit
(S)tatus display
(R)S-232C
Ter(m)inal
(-)Detector temp

MS-D(O)S
(V)ersion
(P)rinter
P(a)sswords

System>
Example of how to use the Thermostat option
System> Current detector temperature: 25.2oC
Detector temperature [oC]
24.0->20
Control detector temperature (Y/N)N->Y
Detector cooling can cause water to condense inside the instrument if the
relative humidity is above 73 % !

When the detectors are cooled, water can condense inside the instrument depending on room
temperature, detector temperature and relative humidity. Please check that the relative
humidity is below the number given in the warning text.
Detector temperature can be selected to be part of programmable output by using the heading
TD.

175

2.32 Thermostat option

176

2.33 Total count rate

2.33 Total count rate

A total count rate register is provided for each of the 7 standard isotopes and one register for
the user definable isotope (called Other). At the end of each assay a total accumulated count
rate or activity for that assay is printed out. Each of these count rates is then accumulated to
form the Total count rate. This is then the total count rate for all assays since the last time the
total count rates were reset. You can display the total count rates, print or reset them. You can
also define efficiencies for each isotope; these are used to transform count rates into activities.
Type T in the Ready or Count state to get to the Total count rate functions. See the example on
the next page.

177

2.33 Total count rate

Count>T
(H)elp
(S)how

(Q)uit
(P)rint

(R)eset

(D)efine efficiencies

Total countrate>S
Total countrate and activity since Wed 12-July-2000 13:53
Isotope
H-3
I-125
C-14
S-35
Cr-51
P-32
P-32C
Other
(H)elp
(S)how

CCPM
32597739
0
124189482
86669
0
0
0
2014554
(Q)uit
(P)rint

DPM
0
0
2978592
0
0
0
0
0

(R)eset

(D)efine efficiencies

Total countrate>D
Efficiency estimates used (Y/N) N->Y
Isotope
1 H-3
2 I-125
3 C-14
4 S-35
5 Cr-51
6 P-32
7 P-32C
8 Other
(H)elp
(S)how

Efficiency
50.00->
0.00->
75.00->90
0.00->
0.00->
0.00->
0.00->
0.00->
(Q)uit
(P)rint

(R)eset

(D)efine efficiencies

Total countrate>S
Total countrate and activity since Wed 12-July-2000 13:53
Isotope
H-3
I-125
C-14
S-35
Cr-51
P-32
P-32C
Other
(H)elp
(S)how

CCPM
32597739
0
124189482
86669
0
0
0
2014554
(Q)uit
(P)rint

Eff
50.0
90.0

(R)eset

DPM
0
0
2978592
0
0
0
0
0

nCi
29367
0
62157
0
0
0
0
0

(D)efine efficiencies

Total countrate>

178

kBq
1087
0
2300
0
0
0
0
0

2.34 Window settings

2.34 Window settings

2.34.1 Radiation Energy
Radioactive isotopes disintegrate spontaneously. The disintegration results in emissions of
alpha, beta or gamma radiation. The radiation type and the energy emitted is specific for each
isotope. In the case of beta radiation the emitted beta particles have a wide range of energies,
the energy spectrum forming a continuous distribution.
Beta-radiation energy ranging from 0 to 2000 keV can be measured in MicroBeta (the most
common beta emitters have their energies between these energy values). The energy scale is
converted into a logarithmic scale and divided into 1024 channels. The energy spectrum of a
beta emitting isotope falls into a certain segment of the energy scale, called an energy window.
A window is a part of the energy scale consisting of many adjacent channels e.g. a window
beginning from channel 5 and extending to channel 360 will cover the energy spectrum of
tritium in MicroBeta.

2.34.2 Window setting

The MicroBeta LSC has built in windows for the isotopes 3H, 125I, 14C, 35S, 33P, 51Cr, 32P, 32P
Cerenkov and for Luminescence counting. The window for Other can be set by the user. The
default value is 5-1024. The preset energy windows shown below are used for these isotopes
when the isotope is chosen in the protocol. When measuring dual labelled samples any of these
window combinations can be used. Note: Before counting, MicroBeta should be normalized for
each isotope used.
Isotope
3
H
125
I
14
C
35
S
33
P
51
Cr
32
P
32
P Cerenkov
Other
Luminescence

Channel settings
5 - 360
5 - 530
150 - 650
150 - 650
150 - 650
5 - 170
250 - 960
5 - 1024
5 - 1024
5 - 1024

If only upper or lower photomultiplier tubes are used, then the default window is 150 -1024 for
non-luminescence counting.

179

2.34 Window settings

180

Part 2J Operating information for

MicroBeta JET
The information in these chapters is for MicroBeta JET only. For the equivalent MicroBeta
TriLux information the chapter with the same number in the preceding part.

181

182

2J.0 Injector setup

2J.0 Injector setup

2J.0.1 Injector modules and channels
When setting up the injectors you are sometimes required to specify the injector module
number(s) and the channel number(s). An injector module is a pump with 1, 2, 3 or 6 syringes
each feeding tubing that goes to the mask adapter which positions the tips of the tubes above
the well(s). Each syringe and hence piece of tubing constitutes one channel. A single detector
instrument can have up to four modules each with one channel. A multiple detector instrument
can have up to two modules, each with as many channels as there are detectors in the
instrument i.e. 2, 3 or 6.

This figure shows the injector system for a six detector MicroBeta JET in which there are two
modules each with six channels.
Each module and channel can be switched on or off and thus needs to be identified in
parameter setting. This is done by giving the number 0 or 1 in the right position to identify the
modules and then 0 or 1, again in the right position to identify the channels. E.g.
Injector module number
Use channels

>1
>100100

would mean that there is a six detector instrument with a single injector module using channels
(syringes) 1 and 4.
Injector module number
Use channels

>2
>111111

This is a six detector instrument with two modules and all six channels of the second module in
use.

183

2J.0 Injector setup

2J.0.2 Selecting Injector setup

The MicroBeta JET injectors need setting up before they can be used. This can be done by
pressing command letter X when the Counting menu is shown. The injector setup menu then
appears as follows:
(H)elp (Q)uit
(I)nit (D)ispense (P)rime
(T)ip
(W)ash Suck-(b)ack Co(m)mand (S)tatus
O(f)f (N)ext pos (G)oto pos (C)learconveyor
Disp(e)nser IN/OUT

2J.0.3 (H)elp
Press H to get help about the various selections.

2J.0.4 (Q)uit
Press Q to quit from the Injector menu.

2J.0.5 (I)nit
Press I to initialize the injector module. You must give the number(s) of the injector module(s)
to be initialized. Power is switched on to the module and the piston position is reset. This must
be the first operation in setup to get the injector system ready.

2J.0.6 (P)rime
This operation fills the tubing with liquid so that there is no dead volume during dispensing.
Make sure the tubing is properly connected before this operation. The input tubing should be in
the reservoir from where the liquid is coming. Press P to begin this operation. It will take a
few seconds. You must give the number(s) of the injector module(s) to be primed and also the
numbers of the channels to be used.

2J.0.7 (T)ip
This operation is to make sure that there are no drops hanging from the tips of the injectors. It
should be performed after prime has been done. Press T to perform it. You must give the
number(s) of the injector module(s) to be tipped and also the numbers of the channels to be
used.

184

2J.0 Injector setup

2J.0.8 Disp(e)nser IN/OUT

This command should be used to move the mask adapter with its embedded injectors into the
counter ready for dispensing to occur. Press E to cause this to happen. This command should
be given before the Dispense command described next.
If the mask adapter is already in position in the counter and you give this command then the
adapter will be moved out to the injector chamber.

2J.0.9 (D)ispense
Press D to actually cause injection to actually occur. This should not be done until Init, Prime
and Tip have been performed. Also the mask adapter should have been moved into place and
the plate should be in position. You must give the number(s) of the injector module(s) to be
dispensed and also the numbers of the channels to be used. Then give the volume to be
dispensed. The units are microlitres. The minimum volume is 10 L which is also the default.
The maximum volume is set by the Dispense volume max. parameter in which the sample
volume in the well before dispensing is also taken into account.
For test purposes you can have the mask adapter out and put a waste tray or an empty plate
under the injectors to catch the liquid.

2J.0.10 Suck-(b)ack
This command is used to empty the tubing after you have completed dispensing. The liquid
will be sucked back into the liquid reservoir, not only emptying the tubing but ensuring that no
liquid is wasted. Press B to begin this operation. This operation should be performed before
Wash is done.

2J.0.11 (W)ash
This command is used for cleaning the tubing after operation has finished and the tubing has
already been emptied of liquid by the Suckback command. Make sure the mask adaptor has
been moved out to the injector chamber and a waste tray has been put under the injectors
before beginning this operation. The input tubing must be in a reservoir of water or if necessary
the cleaning liquid recommended by the manufacturer of the liquid you have been dispensing.
Press W to begin this operation. You must give the number(s) of the injector module(s) to be
washed and also the numbers of the channels to be used. You must also specify the volume of
liquid to be used for dispensing. The units are microlitres and the maximum value is 30,000
L.

185

2J.0 Injector setup

2J.0.12 Co(m)mand
Pressing M allows injector specific macros to be sent to the injector unit. This is mainly for
service purposes.

2J.0.13 (S)tatus
If you want to see the status of the injectors, then press S. If there is an error condition you will
be informed of it. Otherwise you will be told that the system is ready for operation.

2J.0.14 O(f)f
Press F to switch off the injectors.

2J.0.15 (N)ext pos

When you have the mask adapter and a plate in position you can press N to move the plate to
the next position ready for dispensing to the next well.

2J.0.16 (G)oto pos

Press G to move the plate to a selected well position.

2J.0.17 (C)lear conveyor

Press C to move the plate from under the mask adapter back to the cassette rack.

186

2J.1 Beginning operation of MicroBeta JET

2J.1 Beginning operation of MicroBeta JET

The following conventions are followed all through the MicroBeta user interface:

2J.1.1 Menu selections

Menu selections can be entered when the system is in one of the following states:
Ready>
Protocols>
Count>
Info>
System>
Counting protocol>
CPM normaliz. protocol>
DPM standard. protocol>
Assay protocol>
Conveyor>
Total count rate>
Injector>

=
=
=
=
=
=
=
=
=
=
=
=

Ready for next operation

Protocol operations
Counting operations
Additional info.
System level operations
Counting protocol editor
CPM norm. protocol editor
DPM std. Protocol editor
MultiCalc assay protocol operation
Conveyor operation
Total count rate manipulation
Injector operations

Command options available, e.g. those shown below, are displayed before the prompt text, e.g.
Ready:
(H)elp
(C)ount

(I)nfo
(P)rotocols

(S)ystem

Ready>

The command is executed by pressing the letter (either lower or upper case) enclosed inside
parentheses. However commands which include a protocol number (e.g. giving a pure protocol
number in Ready state to start counting of that protocol) must be completed by pressing the
Enter key.
Help text can be obtained by pressing H, h or ?. Exit to a higher level by pressing Q. Pressing
Control-E (the Ctrl and the E keys at the same time) exits always to the Ready state.

2J.1.2 Confirming questions and messages

Quite often MicroBeta displays a message such as:
Press any key when ready or / to exit ->

This kind of message is used to give to the user time to think if this is really the correct
operation and to check that everything is prepared for the operation. Press any key (such as
space bar) when ready to continue. Otherwise the user can cancel the operation by pressing
slash /.
187

2J.1 Beginning operation of MicroBeta JET

Messages such as:

Do you accept the new terminal setting? (Y/N) ->

are used to notify the user that the operation under question can affect the functioning of the
instrument, counting results etc. Press Y=Yes to accept the operation or N=No to cancel it.

2J.1.3 Input of data

The rest of the user interface consists mostly of data input. Data input differs somewhat from
menu selection and confirming questions:
- The default or the current value is usually displayed before an arrow sign ->.
- After typing the new value you must press Enter.
- Press the Enter key alone to leave the default or current value unchanged.
- Either letter H, h or character ? provide help text about the subject under question. Press Enter
after H, h or ?.
- Press slash/ or backslash "Error! Bookmark not defined.\" to exit input mode. In the
protocol editor / exits with saving the protocol and \ without saving it. Elsewhere usually both
cause exit without saving the data.
- Input data can be edited (see below).

2J.1.4 Editing input data

If you make a mistake when entering a new input value you can use the Backspace key
(sometimes labelled as BS or a thick left arrow) or the DEL-key to erase the previous
character. If the input data is quite long this can be tedious. For this kind of situation a simple
keyboard editing facility is provided.
Keyboard editing only uses a few keys so it is easy to learn. The default value or the current
value of input data is used as a template for entering a new value. So if you notice that you
have made a mistake during an early phase of entering a long input (such as output selection in
the protocol editor) you can press Enter to complete entering a new value and then go back to
the same line to correct the mistake.

188

2J.1 Beginning operation of MicroBeta JET

The editing keys are:

<- or Control-B

Go back one character in the template (default or current value).

-> or Control-N

Go to the next character in the template.

E.g. Control-B is entered by first pressing down the Control key (usually labelled with Ctrl)
and then typing letter B (lower case or upper case does not matter). If you keep pressing both
the Ctrl-key and letter B down for a while then the repeat function of the terminal sends several
Control-Bs to MicroBeta.
Note that Control-B and Control-N move only inside the default or the current value (displayed
before ->), not inside a newly typed text.
The cursor control keys (left and right arrows) can sometimes be used instead of Control-B and
Control-N. This does not work with every terminal emulator. The up and down arrow keys can
be used when moving up and down in the protocol editor or you can type Lx to jump to line
number x. Please note that the Enter key must be pressed if making changes to any row.
As an example of keyboard editing, suppose you have entered the following printout selection
(the second line):
Printer output: POS CTIME CCPM1 CCPM1%
-> POS CTIMR COUNTS1 CCPM1 CCPM1%

After you press Enter you will see the text:

Error in output heading : CTIMR

and an arrow -> will appear. Press the Control-N keys. As you do this, you will see the letters
POS CTIMR COUNTS1 etc. appearing. After CCPM1% has been displayed the program will
beep to inform that the end of the current value has been reached. Keep Control-B pressed
down and you will see that characters at the end of output selection start disappearing.
Continue until the cursor blinks just after CTIMR (_ shows the place of the cursor):
-> POS CTIMR_

If you go too far then stop pressing Control-B and press Control-N to make the lost characters
visible again. When the cursor is placed just after CTIMR (as in the example) press Backspace
or the DEL key once and R will disappear. Type the letter E.
-> POS CTIM_

(Backspace or DEL pressed)

189

2J.1 Beginning operation of MicroBeta JET

-> POS CTIME_

(Letter E typed)

Keep pressing Control-N until there is a beep and you get:

-> POS CTIME COUNTS1 CCPM1 CCPM1%

You can now continue typing new headings after CCPM1%. E.g.:
-> POS CTIME COUNTS1 CCPM1 CCPM1% MEAN1 CV1%

Now press Enter to complete editing.

If you later want to remove heading COUNTS1 go to line 31 to get the programmable output
selection. Keep pressing Control-N until you are just past COUNTS1:
-> POS CTIME COUNTS1_

Press Backspace or DEL 8 times so that COUNTS1 and the preceding space disappears:
-> POS CTIME_

Keep pressing Control-N until there is a beep and press Enter:

-> POS CTIME CCPM1 CCPM1% MEAN1 CV1%_

You can also add a new heading afterwards. Go to programmable output selection again. Keep
pressing Control-N until the cursor is just after CTIME:
-> POS CTIME_

Now type space and the text SQP(I):

-> POS CTIME SQP(I)_

Keep pressing Control-N until the program beeps and then press Enter:
-> POS CTIME SQP(I) CCPM1 CCPM1% MEAN1 CV1%_

Note: Keyboard editing works with most of the type (2J.1.3) data input

190

2J.1 Beginning operation of MicroBeta JET

2J.1.5 System operation

The figure following is the flow diagram of the whole MicroBeta operating system. It shows
how the control letters inside parentheses lead to various functions. The functions are described
briefly on the following pages.

191

2J.1 Beginning operation of MicroBeta JET

Flow chart of MicroBeta JET commands (normal level)

Info >
(H)elp
(Q)uit
(U)sage
(C)onsumables
(P)ositions
Ch(a)nges
Cu(s)tomizing
S(y)stem

Ready >
(H)elp
(I)nfo
(C)ount
(P)rotocols
(S)ystem

Injector
(H)elp
(Q)uit
(I)nit
(D)ispense
(P)rime
(T)ip
(W)ash
Suck-(b)ack
Co(m)mand
(S)tatus
O(f)f
(N)ext pos
(G)oto pos
(C)lear conveyor
Disp(e)nser IN/OUT

Count >
(H)elp
(Q)uit
(nn) (count.prot.no)
(?)
(Nnn) (norm.prot.no)
(N?)
(Dnn) (std.prot.no)
(D?)
(A)utomatic counting
Operate con(v)eyor
(T)otal count rate
Injector (X)L

Protocols >
(H)elp
(Q)uit
(C)ounting protocol
CPM (n)ormalization prot.
DPM (s)tandardization prot.
(M)ultiCalc assay protocol

System >
(H)elp
(Q)uit
MS-D(O)S
(E)rror beep
(S)tatus display
(V)ersion
(C)lock
Se(t)up mode
(R)S-232C
(P)rinter
(D)ata drive
Ter(m)inal
P(a)sswords
Customi(z)ing
(-)Detector temp.

Conveyor>
(H)elp
(Q)uit
(O)ff
(C)lear conveyor
Rack (u)p
Rack (d)own
Total count rate>
(H)elp
(Q)uit
(S)how
(P)rint
(R)eset
(D)efine efficiencies
Counting protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
CPM norm. protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
(N)ormalization data
DPM stand. protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
(R)eplot
MultiCalc assay protocol >
(H)elp
(Q)uit
(S)how
(P)rint
(L)ist
MicroBeta A:\ >
MB - Back to Ready >

192

2J.1 Beginning operation of MicroBeta JET

MicroBeta JET commands (normal level) with a brief description of each

Operations in the Ready State
(H)elp
- Display Help text
(I)nfo
- Get information on using the instrument
(C)ount
- Start counting or operate conveyor
(P)rotocols
- Edit counting parameters
(S)ystem
- Edit system parameters
Count state operations are available in the Ready state too.
Operations in the Info State
(H)elp
(Q)uit
(U)sage
(C)onsumables
(P)ositions
Ch(a)nges
Cu(s)tomizing
S(y)stem

- Display Help text

- Back to the Ready state
- View a short user's manual
- View a list of consumables available
- View normalization and standardization positions
- View a list of changes after the previous version
- View information on how to customize the program
- View information about the system

Operations in the Count State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(nn) (count.prot.no)
- Start automatic counting by giving protocol number
(?)
- Start automatic counting by selecting protocol from list
(Nnn) (norm.prot.no)
- Start CPM normalization by giving protocol number
(N?)
- Start CPM normalization by selecting protocol from list
(Dnn) (std.prot.no)
- Start DPM standardization by giving protocol number
(D?)
- Start DPM standardization by selecting protocol from list
(A)utomatic counting
- Start automatic counting using IDs
Operate con(v)eyor
- Conveyor operations
(T)otal count rate
- Show and reset total counts
(M)anual counting
- Start manual counting (only at Test level)
Operations in the Protocols State
(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(C)ounting protocol
- Edit counting protocol
CPM (n)ormalization prot.
- Edit CPM normalization protocol
DPM (s)tandardization prot.
- Edit DPM standardization protocol
(M)ultiCalc assay protocol
- Show and print MultiCalc assay protocol

193

2J.1 Beginning operation of MicroBeta JET

Operations in the Counting Protocol State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(E)dit
- Edit or create and edit a protocol
(nn) (= prot.no)
- Edit a protocol by giving protocol number
(D)elete
- Delete a protocol
(S)how
- Show protocol parameters
(P)rint
- Print a protocol
(L)ist
- Print list of protocols
De(f)ault
- Give default values to a protocol
(C)opy
- Copy one protocol to another
Operations in the CPM Normalization Protocol State
The same operations as in the Counting protocol state, plus:
(N)ormalization data
- Print normalization data
Operations in the DPM Standardization Protocol State
The same operations as in the Counting protocol state, plus:
(R)eplot
- Plot DPM standardization curve
Operations in the MultiCalc Assay Protocol State
Contains only operations (S)how, (P)rint and (L)ist.
Operations in the Conveyor State
(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(O)ff
- Stop conveyor
(C)lear conveyor
- Move cassette to rack and move rack to the middle
Rack (u)p
- Move the rack up one level
Rack (d)own
- Move the rack down one level
Test level contains also:
Remove ca(s)s
(N)ext cass
(P)rev cass
Cu(r)rent cass
Ne(x)t pos
(G)oto pos
C(l)ose detector
Op(e)n detector

- Move cassette to rack

- Move next cassette to conveyor and read ID
- Move previous cassette to conveyor and read ID
- Move current cassette to conveyor and read ID
- Move cassette to the next position
- Move cassette to a specified position
- Close detector block
- Open detector block

194

2J.1 Beginning operation of MicroBeta JET

Next le(v)el
Reset r(a)ck
(I)nit mask
Change (m)ask
(T)est IDs
(F)ocus ID reader
(J)ust read IDs
Print ever(y) ID
Print ra(w) IDs
Print (b)ad IDs
Si(z)e checkpoint
Calib (0)-pos
Calib det bloc(k)
(.)Park det
Shelf (1)..(16)
(*)Calib sensors

- Move rack down to next measurement level

- Move rack to the highest level
- Initialize detector mask (small holes)
- Change detector mask
- Read and print ID of each cassette continuously
- Focus and stop with ID reader on
- Read IDs continuously and show IDs on the screen
- Toggle printing of ID bitmaps in ordinary form
- Toggle printing of ID bitmaps in raw form
- Toggle printing of bad ID bitmaps in raw and ordinary
form
- Move cassette to cassette type checkpoint
- Move cassette between zero point and D06
- Move detector block up and down at mask change
position
- Park detector block for shipping
- Move rack to the specified shelf
- Calibrate sensor offset steps

Operations in the Total Count Rate State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(S)how
- Show the accumulated total count rate and activity
(P)rint
- Print the accumulated total count rate and activity
(R)eset
- Reset the accumulated total count rate and activity
(D)efine efficiencies
- Define counting efficiencies of the isotopes
Operations in the Injector State
(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
(I)nit
- Initialize the selected injector module
(D)ispense
- Volume to be dispensed via the selected tubing of the
selected injector module
(P)rime
- Fill the selected tubing of the selected injector module
(T)ip
- Ensure there is no drop hanging from the selected tubing
of the selected module
(W)ash
- Wash the selected tubing of the selected module with the
selected volume of liquid
Suck-(b)ack
- Suck liquid in selected tubing of the selected injector
module back into bottle

195

2J.1 Beginning operation of MicroBeta JET

Co(m)mand
(S)tatus
O(f)f
(N)ext pos
(G)oto pos
(C)lear conveyor
Disp(e)nser IN/OUT

- Injector specific macros can be sent to the selected

injector module
- Show the status of the selected injector module
- Stop injection
- Move the plate to the next position
- Move the plate to the selected position
- Return the plate to the rack
- Move the mask adapter with injectors (injector assembly)
in or out

Operations in the System State

(H)elp
- Display Help text
(Q)uit
- Back to the Ready state
MS-D(O)S
- Exit to DOS
(E)rror beep
- Set error sound on or off
(S)tatus display
- Set status display on or off
(V)ersion
- Show version number of program and MS-DOS
(C)lock
- Set date and time
Se(t)up mode
- Select Setup mode
Setup mode contains also:
(R)S-232C
(P)rinter
(D)ata drive
Ter(m)inal
P(a)sswords
Customi(z)ing
(-)Detector temp
Test level contains also:
Serial (n)o
Temp cali(b)
Detector/shelf (u)sage

- Set serial port parameters

- Set printer parameters
- Select drive for data saving (results)
- Set terminal parameters
- Print protocol passwords
- Set environment strings to customize the program
- Set thermostat (optional)

- Set instrument serial number

- Edit temperature calibration values
- Set detector usage during counting, set program support
for the ParaLux board and set program support for the
plate ID reader if installed and specify 1/16/32 shelf
model

196

2J.6 Counting control

2J.6 Counting control

2J.6.1 Counting parameters
The counting process is controlled by the following parameters in the counting protocol:
10 Counting time
12 Precision (2 sigma)
41 Counting control
Number of repeats
Number of replicates
Number of cycles
Maximum channel counts (a hidden parameter).
48 Delay between plates

Note: Assay protocol parameters are described in chapter 2.26 MultiCalc.

A CPM normalization protocol or DPM standardization protocol does not include parameter 41
Counting control. Instead the crosstalk normalization and crosstalk standardization protocols
contain counting time for both crosstalk samples and standards, e.g.:
10 Counting time for crosstalk
11 Counting time for standards

2J.6.2 Counting time

Counting will be stopped after the specified counting time (in seconds) has elapsed from the
start of the counting, unless some other conditions have already stopped it. The maximum
counting time is 999 999.9 s (more than 11 days).

2J.6.3 Precision (2 sigma)

The statistical uncertainty in samples is expressed as the standard deviation or 'sigma' value
and is calculated by dividing number one by the square root of the counting value. E.g. 10000
counts gives a 'one sigma %' value of 1%. The 2 sigma % value is two times the one sigma %
value, or 2% for 10000 counts.
In MicroBeta, precision is given as 2 sigma % in a range from 0 to 99.9. Zero means that no
precision test is used.
Make sure that the counting time is set long enough if precision is wanted. E.g. two samples of
1000 and 10000 CPMs will take 10 min and 1 min respectively to achieve a 2 sigma % value
of 2%. Precision is checked once a second. The termination flag in results output will be set to

197

2J.6 Counting control

'PREC' if the precision has been reached. The counting stops if the precision has been reached
in all detectors.

2J.6.4 Number of repeats

To check sample stability or to check instrument performance, the same sample can be counted
repeatedly, up to 999 times. If the repeat value is less than or equal to 5 then results are sorted
and some statistical values are calculated and printed after the sample results.

2J.6.5 Number of replicates

To evaluate sample preparation errors, a number of replicates of a sample can be used. The
results for each sample are output separately. After each group of replicate samples, some
statistical values will be sent to output. The maximum number of replicates is 99.

2J.6.6 Number of cycles

All the sample cassettes in the rack can be counted in repeating cycles. One cycle means
counting of all cassettes in a batch once (i.e. those being counted with one protocol). A batch
ends when the next cassette with a protocol ID is found. When the instrument has counted all
the cassettes in this batch, it moves the rack up until the first cassette in the batch is found.
Then it recounts the cassettes in the batch. Counting of the next batch starts when the cassettes
have been counted the number of times specified by the Cycle parameter. The repeating cycles
can be used to check sample stability. The maximum number of cycles are 99.
If the number of cycles is greater than 1, then you will be asked to give the Cycle delay. This is
in minutes and is the time between when one cycle ends and the next one begins.

2J.6.7 Order of operation of repeated counting

Without using injection
If you have set two or all three of the repeat, replicate and cycle parameters to values greater
than 1 the order of operation is:
First, repeat counting of each sample. The output consists of the individual results for each
repeat count for a sample followed by statistics for the repeat.
Second, statistics for all the replicates of the sample are calculated using the statistics for the
repeat.

198

2J.6 Counting control

This procedure is repeated for all samples in an assay batch. When the end of the batch is
reached, counting starts from the beginning by counting the repeats and replicates again. This
procedure continues for as many times as you have defined for the cycle parameter.
Using injection
When injection is selected it changes the counting procedure so that the positions are dispensed
and counted multiple times, depending on the number of Injector Modules used in the protocol.
Also the background sample measuring (using the Background sample = Y option) is different,
the background is measured for every sample as the first repeat (RP = 0), using the counting
time specified for the background sample.
The following example shows the order in which operation occurs.
Dispense with two modules, background sample selected with two cycles, two repeats and two
plates:
Plate 1
For all coded positions
Background
Count with counting time of background, repeat 0
Module 1
Dispense with Module 1
Wait the delay time of module 1
Count with counting time of module 1, repeat 1
Count with counting time of module 1, repeat 2
Module 2
Dispense with Module 2
Wait the delay time of module 2
Count with counting time of module 2, repeat 1
Count with counting time of module 2, repeat 2
Wait plate delay time
Plate 2
For all coded positions
Background
Count with counting time of background, repeat 0
Module 1
Dispense with module 1
Wait the delay time of module 1
Count with counting time of module 1, repeat 1
Count with counting time of module 1, repeat 2

199

2J.6 Counting control

Module 2
Dispense with module 2
Wait the delay time of module 2
Count with counting time of module 2, repeat 1
Count with counting time of module 2, repeat 2
Wait cycle delay time
Cycle 2
Plate 1
For all coded positions
Module 1
Count with counting time of module 1, repeat 1
Count with counting time of module 1, repeat 2
Module 2
Count with counting time of module 2, repeat 1
Count with counting time of module 2, repeat 2
Wait plate delay time
Plate 2
For all coded positions
Module 1
Count with counting time of module 1, repeat 1
Count with counting time of module 1, repeat 2
Module 2
Count with counting time of module 2, repeat 1
Count with counting time of module 2, repeat 2
Note: dispensing and background counting happens only at first cycle.

2J.6.8 Delay between plates

This parameter can be used to control the interval between when the counting of one sample
plate has finished and the next is started. The range is 0 to 9999 minutes.

200

2J.18 Luminescence counting

2J.18 Luminescence counting

2J.18.1 Introduction
Luminescence assays come in two main forms; those making use of luminescence that has a
short signal (flash luminescence) and those making use of that which emits a long stable signal
(glow luminescence). Both can be measured with MicroBeta, however the short emission
assays require reagent addition, mixing and temperature control. This means that you must use
MicroBeta JET with the injector module(s) installed.
Assays with a more stable luminescence chemistry can be measured with MicroBeta TriLux or
MicroBeta JET without using injection. These luminescence assays are designed to be
completed on a solid support e.g. nylon membrane, glass fibre or coated plate, as well as in
solution and can be directly quantified in a short time.
Due to the high signal, glow luminescence reactions can be measured using single PMTs. An
optional mask adapter can be moved into place to reduce the signal by a factor of 10 if
required.
The traditional microtitration plates are totally opaque (black or white). These plates are
counted with the upper tube and a 1450-105 cassette is used. The filters are counted with the
upper or lower tube and a 1450-104 filter cassette is used.
A special feature when using MicroBeta as a glow-type luminometer is the 1450-466 coloured
sealing tape. The saturation of PMTs is prevented and crosstalk between adjacent positions is
minimized by cutting the intensity of luminescence light with the 1450-466 coloured sealing
tape.
The luminescence units used are LCPS: luminescence counts per second i.e. corrected CPSvalues/100. Before counting the actual samples, the detector and hence the LCPS-values should
be normalized with a luminescence solution as similar to the actual samples as possible.
The normalization is done first and it is made use of when counting the actual samples.

2J.18.2 Preparation of glow-type normalization samples (96 and 384well format)

Make a sample solution which corresponds to the samples to be analyzed, i.e. has the same
enzyme and substrate and pipette the same volume as in the samples, into well/membrane G11
(96-well, 1-6 det.), N22 (384-well, 1-6 det.).
An empty background plate is used for background measurement
201

2J.18 Luminescence counting

The 96-well sample array is as follows, where S1 is the sample solution:

1

A
B
C
D
E
F
G
H

10

11

12

S1

Plate
Close the sample plate with 1450-461 sealing tape or with 1450-466 coloured sealing tape and
place on a 1450-105 cassette for counting.
Filter
Enclose the filter in a 1450-432 plastic sample bag and place it on a 1450-104 cassette; use
1450-466 coloured sealing tape between the sample bag and detector.

2J.18.3 Preparation of glow-type normalization samples (24-well

format)
The procedure is the same as with a 96-position sample support, but the normalization sample
position is D5 for S1.
1
A
B
C
D

S1

2J.18.4 Preparation of flash-type normalization samples (96-well

format)
Standards are arranged in blocks on the normalization plate. If background sample
measurement is used then the background sample must be in the first position in each block on
the plate, see the plate maps following.

202

2J.18 Luminescence counting

The plate maps for different detector configurations with 96-well plates and the maximum
number of replicates are shown below. If there is less than the maximum number of replicates,
the replicates positions are from left-to-right and top-to-bottom.
3 and 6-det. models without background samples. Rows E-H are empty in the 3-det. model.

A
B
C
D
E
F
G
H

1
S1
S1
S1
S1
S4
S4
S4
S4

2
S1
S1
S1
S1
S4
S4
S4
S4

3
S1
S1
S1
S1
S4
S4
S4
S4

4
S1
S1
S1
S1
S4
S4
S4
S4

5
S2
S2
S2
S2
S5
S5
S5
S5

6
S2
S2
S2
S2
S5
S5
S5
S5

7
S2
S2
S2
S2
S5
S5
S5
S5

8
S2
S2
S2
S2
S5
S5
S5
S5

9
S3
S3
S3
S3
S6
S6
S6
S6

10
S3
S3
S3
S3
S6
S6
S6
S6

11
S3
S3
S3
S3
S6
S6
S6
S6

12
S3
S3
S3
S3
S6
S6
S6
S6

3- and 6-det. models with background samples. Rows E-H are empty in the 3-det. model.

A
B
C
D
E
F
G
H

1
B1
S1
S1
S1
B4
S4
S4
S4

2
S1
S1
S1
S1
S4
S4
S4
S4

3
S1
S1
S1
S1
S4
S4
S4
S4

4
S1
S1
S1
S1
S4
S4
S4
S4

5
B2
S2
S2
S2
B5
S5
S5
S5

6
S2
S2
S2
S2
S5
S5
S5
S5

7
S2
S2
S2
S2
S5
S5
S5
S5

8
S2
S2
S2
S2
S5
S5
S5
S5

9
B3
S3
S3
S3
B6
S6
S6
S6

10
S3
S3
S3
S3
S6
S6
S6
S6

11
S3
S3
S3
S3
S6
S6
S6
S6

12
S4
S3
S3
S3
S6
S6
S6
S6

1- and 2-det. models without background samples. Rows E-F are empty in the 1-det. model.

A
B
C
D
E
F
G
H

1
S1
S1

2
S1
S1

3
S1
S1

4
S1
S1

5
S1

6
S1

7
S1

8
S1

9
S1

10
S1

11
S1

12
S1

S2
S2

S2
S2

S2
S2

S2
S2

S2

S2

S2

S2

S2

S2

S2

S2

203

2J.18 Luminescence counting

1- and 2-det. models with background samples. Rows E-F are empty in the 1-det. model.

A
B
C
D
E
F
G
H

1
B1
S1

2
S1
S1

3
S1
S1

4
S1
S1

5
S1

6
S1

7
S1

8
S1

9
S1

10
S1

11
S1

12
S1

B2
S2

S2
S2

S2
S2

S2
S2

S2

S2

S2

S2

S2

S2

S2

S2

2J.18.5 Preparation of flash-type normalization samples (24-well

format)
Plate maps for different detector configurations with 24-well plates and the maximum number
of replicates are shown below. If there is less than the maximum number of replicates, the
replicates positions are from left-to-right and top-to-bottom.
3- and 6-det. models without background samples. Rows C-D are empty in the 3-det. model.

A
B
C
D

1
S1
S1
S4
S4

2
S1
S1
S4
S4

3
S2
S2
S5
S5

4
S2
S2
S5
S5

5
S3
S3
S6
S6

6
S3
S3
S6
S6

3- and 6-det. models with background samples. Rows C-D are empty in the 3-det. model.

A
B
C
D

1
B1
S1
B4
S4

2
S1
S1
S4
S4

3
B2
S2
B5
S5

4
S2
S2
S5
S5

204

5
B3
S3
B6
S6

6
S3
S3
S6
S6

2J.18 Luminescence counting

1- and 2-det. models without background samples. Row C is empty in the 1-det. model.

A
B
C
D

1
S1

2
S1

3
S1

4
S1

S2

S2

S2

S2

1- and 2-det. models with background samples. Row C is empty in the 1-det. model.

A
B
C
D

1
B1

2
S1

3
S1

4
S1

B2

S2

S2

S2

2J.18.6 Normalization protocol

Edit the normalization protocol so that it is suitable for luminescence measurements. To do this
in the Ready-state press P (P)rotocol then N (N)ormalization protocols. Select edit (E) and the
protocol number.
Edit the protocol parameters. See chapters 2.23 and 2J.23 for examples of normalization
parameters. Select luminescence (9) on line 5, isotope 1, and the counting time on (line 10).
Typically this is 1 s. If required, you should also select the mask adapter and if you want
injection then the number of injector modules must be given (i.e. not zero). These are both part
of the settings for parameter 5.
For glow-type luminescence a background plate is counted only if selected on line 42.
For flash-type luminescence using injectors a background plate is not used but instead, if
required, background samples as described in sections 2J.18.4 and 5 above.
Make sure the injectors are ready for operation as described in chapter 2J.0 Injector operation.

2J.18.7 Normalization counting

First place the empty background plate in a cassette (if you are using such a plate)
Place the plate with the normalization samples in a cassette that is identified with the
corresponding normalization protocol number in the protocol number area and with a NORM
label in the function code area.

205

2J.18 Luminescence counting

Insert the cassette(s) into the rack and close the door. Press either A (Automatic counting), or
Nxx, where xx is the normalization protocol number, in the Ready or Count state to start
counting.
If Nxx is selected the program asks:
Do you want to continue counting after normalization? (Y/N)->

If Y (Yes) is selected, the counting continues after counting the normalization plate, if N (No)
is selected the counting stops after the normalization. If normalization has already been done
for the protocol then the program asks first:
This protocol contains normalization data.
Continue? (Y/N) ->

This is to prevent accidental overwriting of normalization data.

If automatic counting is selected the normalization cassette can be placed anywhere in the rack.
The NORM label is needed in this case.
The counting results for the normalization are output as specified on lines 30 - 37 in the
normalization protocol, and after them the background and detector efficiency values are given.

2J.18.8 Counting luminescence samples

Edit a counting protocol:
In the Ready state select P (P)rotocol then C (C)ounting protocols.
Then select edit (E) and protocol number.
On parameter line 2 select 1 (CPM) and on line 3 select the number of the luminescence
normalization protocol defined above. On line 42 select Y (Background correction) and on the
next line Y (Use normalization background). Edit the other parameters normally.
Start counting in the Ready or Count state using automatic counting and a protocol ID label or
start with the protocol number and press Enter.
When counting the samples, the CPS-values are shown on the status display. The stored
normalization factors are used for correcting the LCPS-values in the printout. The normalized
LCPS-values are shown after counting on the status display.

206

2J.23 Normalization

2J.23 Normalization
2J.23.1 What is normalization?
MicroBeta JET has 1, 2, 3, or 6 detectors allowing it to count the respective number of samples
simultaneously. In order for the results from each detector to be equivalent, irrespective of
small variations in efficiency and background between detectors, it is necessary to determine
the relative efficiency and background of each detector and then correct for it. This is called
normalization.
Normalization without injection
In the case when the injection modules are not used, normalization is done by measuring an
optional background plate followed by one or two standard samples with defined activity or
CPM in each detector. Once CPMs have been measured the relative efficiencies can be
calculated. The efficiency of the detector giving the highest count rate is then defined to be one
and the other detector efficiencies are expressed as a fraction of this value. These fractions are
called efficiency coefficients.
If isotope activity is given, then absolute efficiencies are calculated by dividing the count rates
by the activity. In this way, sample quench and detector absolute efficiencies can be corrected
for.
When measuring a sample with a particular detector, the CPM of the sample is corrected by
dividing the CPM by the efficiency coefficient. In dual label counting, corrections are made
using dual label correction formulae (see 3.3 Calculation methods).
When normalization is done the results are stored with the normalization protocol. The
normalization data can be used by one or several protocols and is selected when editing the
protocol (see 2J.25 Protocols).
Normalization with injection
The principle is the same as normalization without injection but there is a difference in the
arrangement of the background and standards. In addition parameter line 5 must be:
9) Luminescence. With other selections the Mask Adapter can be selected but not injection.
Instead of a background plate, a background sample is used for each detector. The position of
the background sample and standards is shown in the plate maps in section 2J.23.4.

2J.23.2 When is normalization necessary?

Each counting or assay protocol requires normalization data to be stored before it can calculate
corrected CPMs for the samples counted. A fresh normalization must be done when a new
207

2J.23 Normalization

isotope is counted or when counting features, such as isotope or window, are changed. When a
new type of microtitration plate is used, crosstalk normalization may be necessary.

2J.23.3 Editing normalization parameters

Normalization parameters are stored in normalization protocols. These parameters are shown in
the figure for the case in which injection is selected i.e. luminescence and mask adapter are
both selected.
Note: If you do not want injection then set the dispensing modules parameter to zero.
Otherwise set the number of dispensing (injector) modules using the code described on the next
page:
Line (1..46, type / to exit, ? for help)
1
1 Protocol name:
->flash lumi norm
Normalization not done
3 Crosstalk correction (Y/N)
N
4 Number of labels:
1)Single 2)Dual
1
5 Isotope 1: 1)H-3
2)I-125 3)C-14
4)S-35
5)Cr-51 6)P-32
7)P-32 Cerenkov
8)Other
9)Luminescence
9
PMT use: 1)Normal 2)Upper 3)Lower 2
Window 1:
5-1024
Use mask adapter (Y/N)
N
Dispensing: 1)High 2)Low 3)Custom
1
Dispensing modules:
11
7 Sample volume [uL]
100
8 Dispensing volume [uL]
10
9 Delay time after dispensing [s]
1.0
10 Counting time (s)
1.0
12 Precision (2 sigma) [%]
0.2
30 Printer output: 1)No
2)Short
3)Long 4)Programmable 3
32 Display output: 1)No
2)Short
3)Long 4)Programmable 3
34 External output: 1)No
2)Short
3)Long 4)Programmable 1
36 File output:
1)No
2)Short
3)Long 4)Programmable 1
40 Change special features (Y/N)
N
41
Number of replicates
1
42
Background sample (Y/N)
N
Background counting time [s]
60.0
43
Half-life correction (Y/N)
N
45
Use password (Y/N)
N
46
Special plate: 1)Wallac 2)BP filter
3)Other
1

208

->

->
->

->
->
->Y
->
->01
->
->
->
->
->
->
->
->
->
->Y
->
->
->
->
->
->

2J.23 Normalization

One module or only the first module

The second of two modules
Both of two modules
The third of three modules
The first and third of three
etc up to
All four of four modules

=1
= 10
= 11
= 100
= 101
= 1111.

There can be a maximum of 100 normalization protocols, numbered from 0 to 99. Protocol
number 0 is the default protocol.
Protocols 91 to 98 inclusive are made at the factory for isotopes 3H, 125I, 14C, 51Cr, 32P, dual
label 3H/14C, 35S, and 32P Cerenkov respectively. These may not be deleted because they are
connected to the corresponding counting protocols. Note that no normalization is done for
those protocols. You must use these protocols to do the normalization before they can be used
for counting unknowns.
Press P and N in the Ready state to choose the normalization protocol state. The protocols can
then be edited, copied, deleted, printed and displayed. Press F to get default parameters and E
to edit a protocol. C is used for copying a protocol including normalization data and D for
deleting a protocol.
Note: the normalization data will be destroyed when editing an old normalization protocol. The
normalization must therefore be done again for this protocol. Only the protocol name and
password can be edited without destroying normalization data.
See chapter 2J.25 Protocols for further details on normalization parameters and editing.

2J.23.4 Preparing the normalization samples

When injection is not used
Depending on which type of MicroBeta you have it is supplied from the factory with:
(for 1-6 detector instruments with 24 or 96-well capability) a 24-well normalization sample
plate containing standards for 3H, and 14C. The 3H standard is in D5 and the 14C standard is in
D6.
The sample positions and the activities for the standards are shown on a label on the plate. The
activity values can be inserted into the normalization protocol.

209

2J.23 Normalization

Be careful that the samples are in the correct positions when making your own normalization
plates. The positions cannot be changed by modifying protocol parameters. In 24-well plates
the standard for isotope 1 is in D5 and the standard for isotope 2 is in D6. Isotope 1 is the lower
energy isotope in dual label counting. In single label counting the standard is in the isotope 1
position, i.e. D5, with one exception:
the standard for 14C must be in the isotope 2 position in single label counting with 24-well
plates.
This is because the factory made normalization plate could be used for both single label 3H and
14
C normalizations and dual label 3H/14C normalizations.
For best results, 96-well plates should be used as normalization plates if 96-well sample plates
are to be counted in a 1-6 detector counter. The isotope 1 standard should be in G11 and the
isotope 2 standard in H12.
See 2.7.2 Crosstalk CPM normalization for crosstalk sample positions and 2J.18 for
luminescence sample positions.
If background sample is selected on line 42, then thermal backgrounds for count subtraction
are measured automatically. An empty background cassette must be the first normalization
cassette followed by the normalization cassette with standards.

210

2J.23 Normalization

See the table below for sample positions in normalization plates:

Sample position in a normalization plate
24-well
96-well
(1-6 det.)
(1-6 det.)
Isotope 1 standard
Isotope 2 standard
14
C single label

D5
D6
D6

G11
H12

When injection is used

Standards are arranged in blocks on the normalization plate. If background sample
measurement is used then the background sample must be in the first position in each block on
the plate, see the plate maps following.
The plate maps for different detector configurations with 96-well plates and the maximum
number of replicates are shown below. If there is less than the maximum number of replicates,
the replicates positions are from left-to-right and top-to-bottom.
3- and 6-detector models without background samples. The rows E-H are empty in the 3detector model.

A
B
C
D
E
F
G
H

1
S1
S1
S1
S1
S4
S4
S4
S4

2
S1
S1
S1
S1
S4
S4
S4
S4

3
S1
S1
S1
S1
S4
S4
S4
S4

4
S1
S1
S1
S1
S4
S4
S4
S4

5
S2
S2
S2
S2
S5
S5
S5
S5

6
S2
S2
S2
S2
S5
S5
S5
S5

211

7
S2
S2
S2
S2
S5
S5
S5
S5

8
S2
S2
S2
S2
S5
S5
S5
S5

9
S3
S3
S3
S3
S6
S6
S6
S6

10
S3
S3
S3
S3
S6
S6
S6
S6

11
S3
S3
S3
S3
S6
S6
S6
S6

12
S4
S3
S3
S3
S6
S6
S6
S6

2J.23 Normalization

3- and 6-detector models with background samples. Rows E-H are empty in the 3-detector
model.

A
B
C
D
E
F
G
H

1
B1
S1
S1
S1
B4
S4
S4
S4

2
S1
S1
S1
S1
S4
S4
S4
S4

3
S1
S1
S1
S1
S4
S4
S4
S4

4
S1
S1
S1
S1
S4
S4
S4
S4

5
B2
S2
S2
S2
B5
S5
S5
S5

6
S2
S2
S2
S2
S5
S5
S5
S5

7
S2
S2
S2
S2
S5
S5
S5
S5

8
S2
S2
S2
S2
S5
S5
S5
S5

9
B3
S3
S3
S3
B6
S6
S6
S6

10
S3
S3
S3
S3
S6
S6
S6
S6

11
S3
S3
S3
S3
S6
S6
S6
S6

12
S3
S3
S3
S3
S6
S6
S6
S6

1- and 2-detector models without background samples. Rows E-F are empty in the 1-detector
model.

A
B
C
D
E
F
G
H

1
S1
S1

2
S1
S1

3
S1
S1

4
S1
S1

5
S1

6
S1

7
S1

8
S1

9
S1

10
S1

11
S1

12
S1

S2
S2

S2
S2

S2
S2

S2
S2

S2

S2

S2

S2

S2

S2

S2

S2

1- and 2-detector models with background samples. Rows E-F are empty in the 1-detector
model.

A
B
C
D
E
F
G
H

1
B1
S1

2
S1
S1

3
S1
S1

4
S1
S1

5
S1

6
S1

7
S1

8
S1

9
S1

10
S1

11
S1

12
S1

B2
S2

S2
S2

S2
S2

S2
S2

S2

S2

S2

S2

S2

S2

S2

S2

212

2J.23 Normalization

Plate maps for different detector configurations with 24-well plates and the maximum number
of replicates are shown below. If there is less than the maximum number of replicates, the
replicates positions are from left-to-right and top-to-bottom.
3- and 6-detector models without background samples. Rows C-D are empty in the 3-detector
model.

A
B
C
D

1
S1
S1
S4
S4

2
S1
S1
S4
S4

3
S2
S2
S5
S5

4
S2
S2
S5
S5

5
S3
S3
S6
S6

6
S3
S3
S6
S6

3- and 6-detector models with background samples. Rows C-D are empty in the 3-detector
model.

A
B
C
D

1
B1
S1
B4
S4

2
S1
S1
S4
S4

3
B2
S2
B5
S5

4
S2
S2
S5
S5

5
B3
S3
B6
S6

6
S3
S3
S6
S6

1- and 2-detector models without background samples. Row C is empty in the 1-detector
model.

A
B
C
D

1
S1

2
S1

3
S1

4
S1

S2

S2

S2

S2

1- and 2-detector models with background samples. Row C is empty in the 1-detector model.

A
B
C
D

1
B1

2
S1

3
S1

4
S1

B2

S2

S2

S2

213

2J.23 Normalization

2J.23.5 Normalization procedure

-First place the empty background plate in a cassette (if you are using such a plate) and then
place the plate with the normalization samples in a cassette that is identified with the
corresponding normalization protocol number in the protocol number area and with a NORM
label in the function code area. Insert the cassette into the rack and close the door. Press either
A (Automatic counting), or Nxx, where xx is the normalization protocol number, in the Ready
or Count state to start counting. If Nxx is selected the program asks:
Do you want to continue counting after normalization? (Y/N)->

If Y (Yes) is selected, the counting continues after counting the normalization plate, if N (No)
is selected the counting stops after the normalization. If normalization has already been done
for the protocol then the program asks first:
This protocol contains normalization data.
Continue? (Y/N) ->

This is to prevent accidental overwriting of normalization data.

If automatic counting is selected the normalization cassette can be placed anywhere in the rack.
The NORM label is needed in this case.
The counting results for the normalization are output as specified on lines 30 - 37 in the
normalization protocol, and after them the background and detector efficiency values are given.
Note! Normalization and unknown samples should have the same quench level in dual label
counting.

2J.23.6 Preset normalizations

There are four preset normalizations available in MicroBeta:
Normalization protocol 1 contains a 3H normalization using ScintiStrip plate 1450-419, dry,
200 L volume with crosstalk correction.
Normalization protocol 2 is the same as 1 but using 125I.
Normalization protocol 3 contains a generally used 3H normalization.
Normalization protocol 4 contains a generally used 14C normalization.

214

2J.25 Protocols

2J.25 Protocols
2J.25.1 Four types of protocols
A protocol is a set of parameters that controls the counting of samples. There are four types:
- CPM normalization protocol
- DPM standardization protocol
- MicroBeta Counting protocol
- MultiCalc Assay protocol
A CPM normalization protocol contains parameters used when doing normalization including
when using the JET features (see chapter 2J.23 Normalization) and a DPM standardization
protocol contains parameters for standardization (see chapter 2.12 DPM Standardization).
Standardization is not used with the JET feature. A MicroBeta counting protocol is used when
counting unknown samples. An assay protocol is the corresponding protocol in MultiCalc (see
MultiCalc). A counting protocol uses either a normalization protocol (in CPM counting) or a
standardization protocol (in DPM counting) to get certain parameters, e.g. isotopes and
window settings. Assay protocols are connected to counting protocols, from which they get
both basic parameters, such as isotopes and counting mode, and special features, such as
background and half-life corrections.
The maximum number of protocols for each type is 100 (0 to 99). Number 0 is the default
protocol, which is used if no protocol number is specified.
Counting protocol number 99 is a so-called GLP (Good Laboratory Practice) protocol. Count
the normalization sample plate supplied with the instrument using this protocol. Do this from
time to time to make sure that the instrument produces consistent results. The background, 3H
and 14C samples are counted with all detectors so background level, CPM values and quench
parameter values can easily be compared. See chapter 3.2 Routine maintenance for more
details.
In this chapter only MicroBeta protocols are described. For further information about assay
protocols see chapter 2.22 MultiCalc and the MultiCalc User manual Protocols.

2J.21.2 Listing protocols

To list or edit protocols, press P and then C for counting protocols, N for normalization
protocols or S for standardization protocols in the Ready state. To select the MultiCalc assay
functions as described in chapter 2.22 press M.
215

2J.25 Protocols

To display the protocol list, in the Protocols menu press S (Show). You will see a two column
list of protocols containing protocol number, name (if given) and isotopes. The list of the
counting protocols also contains counting mode (CPM or DPM) and the number for the
normalization or standardization protocol used. The lists of the normalization and
standardization protocols have a column containing the date when the normalization or
standardization has been done. An equals (=) character indicates that crosstalk correction is
used.
If there are more protocols than there is space for on the display, press Enter to get the next
page of protocols. When you have found the one you want to look at, type the number of that
protocol and press Enter.
Type L (List) to printout the protocol list and P (Print) followed by protocol number and Enter
to print a particular protocol.

2J.25.3 Creating and editing protocols

To create a new protocol with default values, press E (Edit). Choose a protocol number not in
the list and press Enter. The first free protocol number is shown in the menu. Protocols have
already some values in them, either default or old parameters. If you want to set default
parameters, press F (deFault) and select the protocol number.
You will be asked for a line number when starting editing a protocol. Press Enter to start
editing from the beginning of the protocol or type a line number followed by Enter to edit a
certain protocol line. You can always jump between protocol lines by pressing L followed by a
line number and Enter. Type P and press Enter to jump to the previous line. The up and down
arrow keys can also be used.
The value in front of the arrow on each line is the default value. Just press Enter if you do not
want to change this value, or type a new value and press Enter if it must be changed. The CtrlB (Ctrl key held down and the B key pressed) and Ctrl-N keys can be used to move within the
input text; the left and right arrow keys can also be used. See 2J.1.4 Editing input data.
NOTE: Unnecessary lines in a protocol can be hidden to make the editor shorter. Press Ctrl-K
(Ctrl and K keys pressed at the same time) to hide the current line. To reveal a hidden line, go
to the next line and press Ctrl-U. To unhide all hidden lines go to line selection line in the
beginning of the editor and press Ctrl-A.
Press / and Enter when you are finished with your editing and want to save the changes made.
If you want to exit without saving the new values, press \ and Enter.

216

2J.25 Protocols

2J.25.4 Example protocol listing

The example alongside shows a luminescence protocol listing. The parameters are described in
section 2J.25.7.
Counting protocol no: 26 Tue 18-July-2000 10:49
Name: Flash lumi
CPM normalization protocol no:2
Wallac 1450 MicroBeta V4.4 S/N 4500759
1 Protocol name:
Flash lumi
2 Counting mode:
1)CPM 2)DPM
1
3 CPM normalization protocol
4 Number of labels: 1)Single 2)Dual
1
5 Isotope 1: 1)H-3
2)I-125
3)C-14
4)S-35
5)Cr-51
6)P-32
7)P-32 Cerenkov
8)Other
9
PMT use: 1)Normal
2)Upper
3)Lower 2
Window 1:
5-1024
Use mask adapter (Y/N)
Y
Use injector (Y/N)
Y
Dispensing: 1)Automatic
2) Manual
1
Dispensing: 1)High 2)Low 3) Custom
1
Dispensing modules
11
7 Sample volume [uL]
100
8 Dispensing volume 1 [uL]
10
Dispensing volume 2 [uL]
10
9 Delay time after dispensing 1 [s]
1.0
Delay time after dispensing 2 [s]
1.0
10 Counting time 1 [s]
1.0
Counting time 2 [s]
1
12 Precision (2 sigma) [%]
0.2
20 Count all positions in cassette (Y/N)
Y
23 Plate orientation 1)Normal
2)Rotated
3)Mirrored 4)Both
1
30 Printer output: 1)No
2)Short
3)Long
4)Programmable
3
32 Display output: 1)No
2)Short
3)Long
4)Programmable
3
34 External output: 1)No
2)Short
3)Long
4)Programmable
1
36 Fileoutput:
1)No
2)Short
3)Long
4)Programmable
1
40 Change special features (Y/N)
Y
41 Counting control (Y/N)
Y
Number of repeats
1
Number of replicates
1
Number of cycles
1
42 Background correction (Y/N)
Y
Use normalization background (Y/N)
N
Background sample (Y/N)
Y
Bgnd counting time [s]
1.0
43 Half-life correction (Y/N)
N
45 Use password (Y/N)
N
48 Delay between plates [m]
0

217

2J.25 Protocols

2J.25.5 Protocol copy and delete

The contents of one protocol can be copied to another by pressing C (Copy). Select the No. of
the protocol to be copied and enter the new protocol number. If you want to delete a protocol,
press D (Delete) and select the protocol number. Press Q (Quit) to return to the Ready state.

2J.25.6 Protecting protocols

Protocols can be protected against unauthorized editing by using passwords. The password is
selected on line 45 in the protocol editor. This prevents other users from changing or deleting
your protocol, but they can still use it (e.g. listing, copying and counting with it). The
MicroBeta supervisor has the authority to list the passwords.

2J.25.7 Parameters in a counting protocol

1. Protocol name
Type a name for the protocol, maximum 170 characters (only the first 16 characters will be
shown in the protocol list) and press Enter. The name may not start with characters ?, / or \.
2. Counting mode
Type 1 for CPM counting or 2 for DPM counting.
3. CPM normalization protocol / DPM standardization protocol
Type the number for the normalization (in CPM counting) or standardization protocol (in DPM
counting) used. Type H or ? to get a list of normalization/standardization protocols. Isotopes
must be entered if no protocol is selected, otherwise the counting protocol uses the same
isotope(s) as in the norm./std. protocol. Type - if no protocol is needed. This means that no
normalization correction is done in CPM counting and no DPMs are calculated in DPM
counting. Lines 4 and 5 are displayed only if no normalization or standardization protocol
number is selected. Information about whether norm./std. has been done is given after line 3.
4. Number of labels
Type 1 for single label counting (one isotope, default) or 2 for dual label counting (two
isotopes).

218

2J.25 Protocols

5. Isotope 1
Select an isotope, in dual labelled samples the isotope with the lower energy. The alternatives
are:
1) 3H (default) 2) 125I
3) 14C
4) 35S
5) 51Cr
6) 32P
32
7) P Cerenkov 8) Other
9) Luminescence
You will be asked for window limits if you select 'Other', otherwise window settings for the
selected isotope are shown.
ParaLux counting can be selected in single label non-crosstalk and non-luminescence
protocols. If ParaLux is selected then AQP(I) is used instead of SQP(I). ParaLux counting has
two modes: High efficiency and Low background. The discriminator channel that is used to
calculate AQP(I) and High efficiency counts can also be optimized.
For PMT use select normal coincidence counting, or use only upper or lower photomultiplier
tubes. Upper is the default if luminescence counting is selected.
Window 1 shows what the window setting is. If 'Other' has been selected as isotope 1 enter the
limits for window 1 in a range from 1 to 1024.
Window 3 is displayed only if 'Other' has been selected as isotope 1 or isotope 2 in dual label
counting. Enter the limits for window 3 in a range from 1 to 1024 or 0 - 0 if no third window is
used.
Use mask adapter - if you select Y for this then the mask adapter will be used for the counting.
Note: this must be set to Y if you want to use the injector because the injector uses the mask
adapter. If you select N the parameters lines following as far as parameter 12 will not appear.
Use injector - select Y if you actually want to use the injector system. If you select N then even
though the mask adapter with injectors is in position, no injection will be done. In the latter
case lines from this one as far as parameter 12 will not appear.
Dispensing - you should select automatic dispensing. The other option, Manual is not available
at present.
Dispensing - you can select the dispensing speed to be high, low or a custom speed set in the
system settings.
219

2J.25 Protocols

Dispensing modules - here you give the number of modules to be used. If you have a single
detector instrument you can have up to four injector modules. If you have 2, 3 or 6 detectors
you can have one or two modules. In order specify the modules used use the following codes:
One module or only the first module
The second of two modules
Both of two modules
The third of three modules
The first and third of three
etc up to
All four of four modules

=1
= 10
= 11
= 100
= 101
= 1111

6. Isotope 2
Displayed only in dual label counting. The higher energy isotope is selected. The alternatives
are the same as for parameter 5 with 14C as default isotope.
7 Sample volume
Displayed only if Use injector has been selected to be Y. This parameter defines the original
amount of sample in the well before dispensing. This is a mandatory entry and it is used for
checking that the dispensed volumes do not exceed the maximum allowed volume. The value
is an integer number in micro litres (L). The minimum value is 0. The default value for this
entry field is empty.
8. Dispensing volume
Displayed only if Use injector has been selected to be Y. This parameter defines the amount of
liquid to be dispensed. The value is an integer number in microlitres (L). The minimum value
is 10 L. The default value is 10 L. There is a separate parameter for each module used. In a
single detector counter this can be up to 4 modules and in multiple detector counters, up to 2
modules.
9. Delay time after dispensing
Displayed only if Use injector has been selected to be Y. The Delay Time defines the time to
wait after dispensing, before the counting starts. There is a separate parameter for each module
used. The resolution is 0.1 seconds. The default value is 0.1 second.
10. Counting time
The counting time is given in seconds. The minimum value is 0.1 and the maximum is 999
999.9 seconds. The default value is 60.0. If Use injector was selected for paramer 5 then there
will be as many counting time parameters as there are dispensing modules.

220

2J.25 Protocols

12. Precision (2 sigma)

Counting stops when the standard deviation is below the two sigma value in all detectors.
Precision is given in percent in a range from 0 to 99.9. 0 means that no precision limit is used.
The default value is 0.2 %.
20. Count all positions in cassette
Press Y (Yes) if you want to count the whole cassette (default) or N (No) if you want to specify
the positions to be counted. If No has been selected the next parameter will appear:
21. Cassette type
Type 1 if you use cassettes with 96 wells (default) or 2 if the cassettes have 24 or 384 wells.
22. Active rows
If you answered No on line 20 you will have to specify the rows to be counted for the first
cassette by typing the letters for these rows. E.g. 'AC' means that samples A1 to A12 and C1 to
C12 will be counted. For a more detailed specification, i.e. giving the positions instead of rows
only, press R (Row coding). In this case specify which positions are to be counted for each
row. The positions are in order from 1 to 12 (or 6 for 24-well plates). Type 1 to count a
position or 0 to skip it. E.g. Row A: ->0011110 means that positions A3, A4, A5 and A6
should be counted. You do not have to enter all the zeros at the end of the line; the row end is
filled with 0 or 1 according to the last typed character. See the following example:
20 Count all positions in cassette (Y/N)
Y
21 Cassette type: 1)96 2)24
1
22
Cassette 1:(96)
Rows:
AB
Row A: 111111111111
Row B: 001111000000
Row C: 000011000000
Row D: 000000000000
Cassette 2:(96)
Rows:
ABCDEFGH
Cassette 3:(96)
Rows:
ABCDEFGH

->N
->
->R
->0011110
->0000110
->0
->AB
->/

An alternative method is to type the column numbers e.g. 5,6,7 equals 000011100000, 1,
equals 100000000000 and 3,, equals 001111111111.
After pressing Enter you will be asked for positions for the next row. The default values are the
same as you have given on the previous row.
Press / when you want to stop specifying counting positions. The rest of the rows will then be
counted as the last edited row. In the example above, the following positions will be counted
for cassette 1: A3, A4, A5, A6, B5 and B6. The following cassettes can then be coded in the
same way as the first one. Press / once again to exit cassette coding. If there are more cassettes

221

2J.25 Protocols

in a batch than there are defined cassettes, the rest of them are counted according to the last
cassette specification.
W can be pressed to code additional cassettes later.
X can be pressed to change the cassette type (96 to 24 (or 96 to 384) and vice versa) for
cassettes 2 to 16, see the example:
20 Count all positions in cassette (Y/N)
Y
21 Cassette type: 1)96 2)24 3)12
1
22
Cassette 1:(96)
Rows:
ABCDEFGH
Row A: 111111111111
Row B: 001111000000
Row C: 000011000000
Row D: 000000000000
Cassette 2:(96)
Rows:
ABCDEFGH
Cassette 2:(24)
Rows:
ABCD
Cassette 3:(24)
Rows:
ABCD

->N
->
->R
->3,4,5,6
->5,6
->0
->X
->AB
->/

23 Plate orientation
Select 2 if rotated plates (A1 at the bottom right corner) are used, 3 if mirrored (A1 at the top
right corner), and 4 if both rotated and mirrored (A1 at the bottom left corner).
30. Printer output
There are four printer output alternatives:
1) No
2) Short
3) Long (default)
4) Programmable
1) No
If No is selected no results are printed.
2) Short
If Short is selected, sample results are printed using the short results format: POS, CCPM1,
CCPM1%. In dual label CCPM2 and CCPM2% are added. CCPM is replaced with DPM in DPM
counting. In luminescence counting it is POS, LCPS and LCPS%.
3) Long
The long output contains: PROT, POS, CTIME, SQP(I) (or AQP(I)), CCPM1, CCPM1%,
plus CCPM2, CCPM2% in dual label counting. In DPM counting DPM1 and DPM1% are added. In
luminescence counting the output is PROT, STIME, POS, CTIME, CPS1, LCPS, LCPS%.

222

2J.25 Protocols

4) Programmable
The output items and arithmetic expressions are defined separately on the next line. Sample
quality monitors can also be selected in the programmable output. Press ? or H to get a list of
output items, monitors and rules for using own formulae. See also chapters 2.1 'Beginning
operation', 2J.23 'Results', 2.29 'Statistics' and 2.21 'Monitors and flags'.
If SPECTRA is selected, three more lines are shown:

The first asks for the energy scale to be selected. It can be logarithmic with the units in
channels or linear with the units in keV or both of these. Next you are asked to specify the
range of the plot. This will be channels or keV depending on your previous selection. Finally
you are asked for the spectrum size, small, medium or large. A smaller size will give faster
output. The two figures following illustrate the type of plots you may get.

Spectrum with logarithmic scale

Spectrum with linear scale

223

2J.25 Protocols

32. Display output

This output is selected when you want to display results on the screen. It can be chosen
independently of the printer output.
Display output has the same output alternatives as the printer output.
The default programmable output, PRINTER means the same data as specified for printer
output.
34. External output
This output is selected when sending data to an external device such as a mainframe computer
connected to the MicroBeta via port 2. The data is in ASCII format (text files).
The output alternatives are the same as in printer and display output, except that alternative 1,
No output, is the default value.
The default programmable output, PRINTER means that the same data is specified as for printer
output.
36. File output
The counting results can be stored on a floppy disk or hard disk on the PC or MicroBeta, or on
the server of a local area network (LAN) (section 2.8 Datafiles). The default setting is 1, no file
output. The alternatives are the same as in printer output.
The default programmable output, PRINTER means that the same data is specified as for printer
output.
The result files are named so that the first part of the name (the part before the period) is the
protocol number and the second part (after the period) is a running number indicating the
assay's execution order. E.g. the first file for protocol 5 is named 5.001, the second 5.002, etc.
Result file names for normalizations begin with N (e.g. N1.001) and standardizations with an S
(e.g. S1.001).
The disk drive (A:, B: for floppy disks, C: and D: for hard disks, F: etc. for LAN drives) and
the path (e.g. \MB\DATA\) for the result files are chosen after the File output. This over-rides
any path set in System mode. The drive and path defined in the System state is the default
value (see chapter 2J.30.9 System/Data drive).

224

2J.25 Protocols

This is the terminal PC path. If results should be stored on the MicroBeta disk, then the path
must begin with an @, e.g. @B:\DATA\.
If a VT52 or VT100 terminal is used then results are stored on the protocol disk as default
(directory B:\RESULTS), see 2.30.10 Terminal.
The file name extension (the part after the period) can be changed on the next line. This must
be a number in the range 1 to 999.
Note: this line is not available if UltroTerm is used.
40. Change special features
Press Y to change special features. These are counting control, background, half-life and
chemiluminescence correction, password and special plate type.
41. Counting control
This and the following questions will be displayed only if Y has been selected on the previous
line.
Press Y to change the counting control parameters. These are number of repeats, replicates and
cycles.
If N is selected then these parameters have no effect.
The number of repeats is the number of times each sample will be counted. The minimum
value is 1 (default) and the maximum is 999. If repeat is greater than 5, results are not sorted
and statistical results are not printed. Repeats cannot be selected in crosstalk.
The number of replicates specifies how many replicates there are for a sample. This is used for
evaluating sample preparation errors. The minimum value is 1 (default) and the maximum is
99.
The number of cycles is the number of times a batch of sample cassettes will be counted. The
minimum is 1 (default) and the maximum is 99. If the number of cycles is greater that 1 then
you will be asked to give the Cycle delay. This is in minutes and is the time between when one
cycle ends and the next one begins.
See chapter 2J.6 Counting control for more information.

225

2J.25 Protocols

42. Background correction

Press Y to use background correction. You can choose between using the background
measured in normalization, giving the position for a background sample or giving a value to be
subtracted from the sample CPM. Answer Yes to the background sample question if you want
to use a background sample. Enter the position for this sample. H12 (96-well, 1-6 detectors),
G12 (12-detectors), D6 (24-well) or P24 (384-well, 1-6 detectors) is the default. Enter also the
counting time (60s is the default). Note that the background sample must be in a position
reached by every detector (see chapter 2.9 Detectors for these positions). The measured
backgrounds are subtracted from CPMs when calculating CCPMs.
Answer No to the background sample question if you want to give a background value. The
value, given in CPM, is subtracted from the sample CPM for an isotope. Two values are given,
one for each window, if dual label counting is used. In dual label DPM counting, three values
are given. The value is in the range from 0.0 (default) to 99.9 CPM or 999 999.9 LCPS.
Note: in the normalization protocol press Y if you want to count the background plate.
43. Half-life correction
Press Y if you want to use half-life correction. The half-life for the isotope is displayed, e.g.
107 500 hours for 3H. No half-life value is given for 14C because it is so long. Note that you
must always specify the half-life if you have selected an isotope of type 'Other'. It must be in a
range from 0 (no half-life) to 999 999.99 hours (114 years).
If normalization or standardization protocol is selected on line 3 then half-life is as specified in
this protocol and it cannot be changed in the counting protocol
Press Y on the next line if you want to set zero time, otherwise the activity is corrected to the
time when counting of the first sample of the assay was started.
The zero time consists of date and time. First enter the date in the format DD-MMM-YYYY, where
DD is the day number (1 - 31), MMM the month in letters (JAN, FEB, MAR, APR, MAY, JUN,
JUL, AUG, SEP, OCT, NOV and DEC) and YYYY the year, e.g. 15-JUN-2000. The date can
also be in the format YYYY-MM-DD, e.g. 2000-06-15. Then enter clock time in the format
HH:MM:SS, where HH is hours in the 24 hour mode (0 - 23), MM minutes (0 - 59) and SS seconds
(0 - 59), e.g. 14:05:30. It is not necessary to give seconds. The default value for zero time is
the current time. See also chapter 2.14 Half-life.
44. Chemiluminescence correction
Press Y to use chemiluminescence correction, otherwise N (default). This correction is done
with a correction formula, see chapter 3.3 Calculation methods.

226

2J.25 Protocols

45. Password
Press Y to select a password if you want to prevent other users from changing your protocol.
Enter the password, maximum 12 characters. This will NOT be echoed to the screen. You will
also be asked to retype it to prevent typing errors. The protocol cannot later be edited or deleted
without knowing the password. You must therefore try to remember it. Only the MicroBeta
supervisor has the authority to list the passwords (see chapter 2J.30 System/Passwords). He or
she can help you if you have forgotten your password.
46. Special plate
Select Special plate type. Wallac is the standard plate. Betaplate filtermat plates and other 24well plates have different dimensions and are counted using just one detector. However, it is
possible to count other 24-well plates with all detectors. This selection comes after the plate
dimension questions. For Betaplate filtermats you can select the 96-well plate format output.
This is a composite of four 24-well plate format outputs. If you select the Other option for plate
type, you must give the plate dimensions. These are the distances of the centres of the A1, A6
and D1 wells from the plate sides. Special cassettes must be used for Betaplate filtermat plates
and other 24-well plates. The default dimensions 17.3/13.4/113.6/71.5 are for the Costar plate
type. For the Falcon plate type the dimensions 14.5/13.4/110.7/71.5 should be used.
The default dimensions for a special 384-well plate are 11.8/9.0/115.3/76.5. These are the
distances of the centres of the A1, A24 and P1 wells from the plate sides.
Note: this line is not available if 96-well cassette has been selected elsewhere in the protocol.
47. Autoquench correction
Automatic quench correction can be done in single label CPM counting. Isotopes must be 1, 2,
3, 4 or 6, and a non-crosstalk CPM normalization protocol must be selected.
48 Delay between plates
The delay between the end of plate measurement and the start of the next plate measurement
can be specified. The range is 0 to 9999 minutes.

2J.25.8 Parameters in a normalization protocol

The parameters are much the same as in the counting protocol with some exceptions.
Information whether normalization/standardization has been done is given after the protocol
name. The following counting protocol parameters are not included in the normalization
protocol:

227

2J.25 Protocols

2.
3.
20.
21.
41.

Counting mode
CPM normalization/DPM standardization protocol
Count all positions in cassette
Cassette type
Counting control

The following parameters are new or changed when compared with the ones in the counting
protocol:
3. Crosstalk correction
Press Y to use crosstalk correction otherwise select N. see chapter 2.7 Crosstalk correction for
further information.
10. Counting time for background
The counting time for the background and crosstalk counting samples in crosstalk counting is
given in seconds. The minimum value is 0.1 and the maximum is 999 999.9 seconds. The
default value is 60.0.
11. Counting time for standards
The counting time for the standard samples is also given in seconds, from 1 to 999 999.9 with
60.0s as the default value.
41. Isotope activity setting
Press Y to set activities for normalization samples. This is isotope dependent and given in
DPM. The range is from 1000.0 to 999 999.9. The default value is 200 000 for isotope 1 and
100 000 for isotope 2.
If activities are given, absolute normalization is done with efficiencies as count rates divided
by the given activity.
If N is selected, relative normalization is done with efficiencies as a fraction of the highest
detector count rate.
47. Strip plate
Press Y if you use plates where crosstalk is different in the X and Y directions. The crosstalk
sample in G10 is counted in this case.
See chapter 2J.23 Normalization for further information.

228

2J.25 Protocols

2J.25.9 Parameters in a standardization protocol

The parameters are much the same as in the counting and normalization protocols with some
exceptions. JET features are not available with standardization but the mask adapter can still be
used. Information whether normalization/standardization has been done is given after the
protocol name. The following counting protocol parameters are not included in the
standardization protocol:
2.
3.
20.
21.
41.
42.

Counting mode
CPM normalization/DPM standardization protocol
Count all positions in cassette
Cassette type
Counting control
Background correction

The following parameters are new or changed when compared with the counting protocol:
2. Sample type
Select normal or scintillation proximity assay for sample type. ParaLux counting is set
automatically if SPA is selected.
3. Crosstalk correction
Press Y to use crosstalk correction otherwise select N. See chapter 2.7 Crosstalk correction for
further information.
14. Easy DPM
Press Y to use Easy DPM quench curves. Only the two DPM normalization samples are
counted in standardization.
20. Number of standards
Give the number of DPM standards used for standardization. The value must be in a range
from 2 to 12 (6 in dual label) with 6 as default.
21. Isotope 1 activity
Give the isotope activity in DPM. The range is from 1000.0 to 999 999.9 with 200 000 as the
default value.
23. Isotope 2 activity
Give the Isotope 2 activity in DPM. The range is from 1000.0 to 999 999.9 with 200 000 as the
default value.

229

2J.25 Protocols

24. Standard curve fit selection

Press Y to use standard curve fitting, otherwise N. If No is selected, smoothing spline with
automatic smoothing will be used or linear interpolation if SPA is selected for the sample type.
25. Curve fit method
This line is displayed only if you have selected Yes on the previous line. The methods for
standard curve fitting are:
1) Smoothing spline (default)
2) Interpolation spline
3) Linear interpolation (default for SPA)
4) Linear regression
Select a method by typing the number and Enter. If you have selected the first alternative,
smoothing spline, you will be asked if the method is to include automatic smoothing. Press Y
(default) if so. Otherwise press N and you will be asked for a smoothing parameter. Enter a
value between 0.0 (no smoothing) and 1000.0 (maximum smoothing), with 1.0 as default.
26. Edit standard curve
Press Y to edit the quench parameter, efficiency and weight values for a standard curve or to
delete curve points.
41. Adjusted activities
Press Y if you want to adjust the activities for standards, otherwise the activity given on line 21
will be used for every standard. After pressing Y you will be able to change activities for the
same number of DPM standards as given on line 20. You can also change the activities for the
two Easy DPM standards. The range is from 1000.0 to 999 999.9 DPM. The value given on
line 21 is default. See chapter 2.12 DPM Standardization for further information.

230

2J.25 Protocols

2J.27 Robotic loading interface

The robotic loading interface is a special door/adapter system that brings a cassette outside so
that a robotic arm can change the plate.
In the MicroBeta program the robotic loading interface is selected as follows:
(S)ystem
(L)evel
(T)esting
Detector/shelf (u)sage
Use shelves: 1)1 2)16 3)32 ->1
(L)evel
(N)ormal counting

The operation is controlled using the RS-232C port 2 (external port) of MicroBeta. When the
cassette is outside, MicroBeta sends the text Ready and carriage return / line feed. The possible
commands are:
H or ?
Nxx
Dxx
xx
A

- list commands
- start normalization using protocol number xx
- start standardization using prot. number xx
- start counting using protocol number xx
- start counting using IDs.

All Ready commands must be followed by a carriage return / line feed or #. Counting can be
interrupted with O. After a plate has been measured and the cassette is outside, MicroBeta
sends the text Continue and carriage return / line feed. At this point O stops counting and any
other character continues counting.
If plates are counted in the inverted position (A1 at the bottom right corner) then Plate
orientation "Rotated" should be selected in the counting protocol. This selection reorders
results so that outputs POS and SEQ are correct.
Warning
The mask adapter has to be moved manually into the counting chamber before starting
counting.

231

2J.27 Robotic loading interface

232

2J.30 System

2J.30 System
2J.30.1 Selecting System
System settings control the general way MicroBeta works - its defaults, hardware connections
etc. To enter the System state press S in the Ready state. You are then given a number of
options as example 1 shows. These are described in more detail here.
Example 1
(H)elp
(C)ount

(I)nfo
(P)rotocols

(S)ystem

Ready>S
(H)elp
(E)rrorbeep
(C)lock
(D)ata drive
Customi(z)ing

(Q)uit
(S)tatus display
(R)S-232C
Ter(m)inal
(I)jector module

MS-D(O)S
(V)ersion
(P)rinter
P(a)sswords

System>

2J.30.2 (H)elp
Press H to get help about the various selections.

2J.30.3 (E)rror beep

User or instrument error normally causes a beep. This can be switched off by selecting N.
Example 2
System>E
Error beep (Y/N) Y ->N

2J.30.4 (S)tatus display

Normally when counting is occurring its status is shown on the Terminal PC screen (see
chapter 2.5 Counting for more details). The counting status display can be switched off or only
currently counted positions can be shown.
Example 3
System>S
Status display:

1)No
2)Current positions
3)Whole plate

233

3->

2J.30 System

2J.30.5 (V)ersion
Show the program and MS-DOS version numbers.
Example 4
System>V
MicroBeta program V5.0. Copyright (C)
Wallac Oy, 1998. All rights reserved.
MS-DOS V3.2. Copyright (C) Microsoft
Corporation, 1987. All rights reserved.

2J.30.6 (C)lock
This allows the date and time to be set. MicroBeta has a calendar clock with battery back-up so
you do not have to set it each time the power has been switched off, normally only when the
instrument is installed. Details of setting the clock are given in chapter 2.3 Clock. When you
set the clock the Terminal PC date and time are also set if GenTerm is used.
Example 5
System>C
Date 12-Jul-2000 -> 13-Jul-2000
Time
14:44 -> 9:30
Set clock(Y/N)->Y

2J.30.7 (R)S-232C
There are three RS-232C communication ports which can be used:
1 for the Terminal PC
2 for an external computer
3 for injector module communication
You can specify the communication parameters for each of these ports separately. Give the
number of the port and press Enter.
The first parameter that can be changed is the speed of transferring the data, the Baud rate. The
possibilities are: 300, 600, 1200, 2400, 4800 or 9600.

234

2J.30 System

The default value for the terminal and external ports is 9600. For the printout port it is 4800. If
these values are not suitable then select the ones you need.
Next is the parity selection, you can have None, Odd or Even where None is the default.
The number of data bits can be 8 or 7 where 8 is the default.
The number of stop bits can be 2 or 1 where 2 is the default.
Lastly you can select the kind of checking process for the data transfer, the handshaking. It can
be None, DTR or XON/XOFF where the default is DTR.
Confirm the selection of these parameters by typing Y twice. When you have done this the
program prints the original and the new settings and returns to the System state if serial port 2
or 3 setting is in question. However when changing the terminal port (port 1) setting the
program then shows a help text that informs you to set your terminal to the same settings which
were selected for MicroBeta before confirming the new setting. When ready press any key. The
program should then ask:
Do you see this text (Y/N)?

If the text appears it is quite sure that the communication works and you can press Y to confirm
the change of terminal port setting. If the text does not appear it is obvious that your new
terminal and MicroBeta serial port settings differ. In that case check from the printer the new
MicroBeta setting and modify your terminal setting if required until you see the above text. If
you cannot make the communication work you have still one possibility. Set the terminal to the
original serial port setting. Switch off the counter and switch on again. Restarting MicroBeta at
this phase returns the original terminal port communication setting. Note: You can lose
communication with the counter also by changing the serial port setting of your terminal alone.
Usually then some strange characters appear around the screen and commands do not function.
To get communication functioning again you can change terminal settings in the following way
if you do not remember the terminal port setting of MicroBeta:
- Try different baud rates. Press e.g. space bar after setting each baud rate.
- If there is no success then change the number of data bits (usually 7 or 8 bits come into
question) and try different baud rates again.
- Usually parity and handshake differences should not prevent normal manual terminal use.
- Another method is to switch the counter off and on again.
235

2J.30 System

The correct parameters (terminal type, baud rate and data bits) are set automatically after
instrument restart. If they differ from the current parameters then a text like the following is
shown: Incorrect terminal baud rate. 9600 bps used. Please set terminal RS232C parameters in (S)ystem /(R)S-232C.
When it seems that communication is working, go to the MicroBeta System-mode and check
the serial port 1 settings and make sure that they are identical with settings of the terminal.
Example 6
System>R
RS-232C port number
1->
Baud rate: 1)300 2)600 3)1200
4)2400 5)4800 6)9600
6->
Parity: 1)None 2)Odd 3)Even
1->
Data bits
8->
Stop bits
2->
Handshake: 1)None 2)DTR 3)XON/XOFF 2->
Set terminal port parameters
(Y/N)->Y
Are you sure?
(Y/N)->Y
Do you see this text (Y/N)?Y

Note: for serial port 3, the connection to the injector modules, the parameters must be: Baud
rate = 6, Parity = 1, Data bits = 8, Stop bits =2 and handshake = 1.

2J.30.8 (P)rinter
Set the printer type and the number of characters per inch. The options are: IBM or Epson FX
and the number of characters per inch 10, 12 or 17. The defaults are Epson FX and 12 cpi.
Example 7
System>P
Printer type:
1)IBM 2)EpsonFX
Printer charac.per inch: 1)10 2)12

3)17

2->
2->

2J.30.9 (D)ata drive

Select diskette or hard disk drive of the terminal PC or MicroBeta on which the results are to
be stored (when using UltroTerm define the result file path in UltroTerm Result filing options).
The result file path can also be given in each protocol.
Example 8
System>D
Data drive:
Path name:

1)A:

2)B:

3)C: 4)Path
4->
C:\MB\RESULTS\->

236

2J.30 System

The options are:

1 = diskette drive A. Select this if you have:
(a) a single drive terminal PC (no other possibilities)
or
(b) a hard disk PC but you want to store results on the diskette (e.g. to be transferred
somewhere else).
2 = diskette drive B. Select this if you have a dual diskette drive terminal PC. Then terminal
emulator software can be kept in drive A all the time (e.g. to enable automatic restart after
power failure).
3 = hard disk drive C. Select this if your terminal PC is equipped with hard disk and you want
large storage capacity. If the GenTerm terminal emulator is used, then results will be stored
into the directory which was active when GenTerm was started.
4 = full disk drive and directory path. Use this if options 1-3 do not match your
requirements. E.g. if you want to store results:
(a) on a hard disk drive other than C (e.g. D),
(b) on a local area network (LAN) connected to the terminal PC,
(c) in a specific directory
or
(d) on the MicroBeta drive.
If 4 is selected then the program asks for the full drive and directory path. Here understanding
of MS-DOS drive and directory concepts is needed. The directory path given here will be used
as such without any checking and modifications in front of the file name. Note that the
directories must exist before any data can be sent to them. Use the command MKDIR in DOS
to make new directories.
Examples (referring to examples a-d above):
(a) Path name: D: ->
- Text D: will be inserted in front of result file name. E.g. D:2.003 , the file will be stored in the
active directory on drive D.
(b) Path name: F: ->
- Here it is supposed that drive F: belongs to the LAN server. Text F: will be inserted in front
of the result file name. E.g. F:17.002 , the file will be stored in the active directory of the LAN
server drive F.

237

2J.30 System

(c) Path name: C:\MB\RESULTS\ ->

- Text C:\MB\RESULTS\ will be inserted in front of result file name. Note that if you specify an
exact directory path for the result files then the path name must end with backslash \. For
example the path C:\MB\RESULTS\3.023 stores result file 3.023 into the directory
\MB\RESULTS\ of hard disk drive C.
(d) Path name: @B:\DATA\->
- Results are stored on the MicroBeta protocol disk B in directory DATA. The character @ at the
head of the path shows that the MicroBeta disk is to be used for storing the results.

2J.30.10 Ter(m)inal
Select the terminal emulation (see chapter 2.31 Terminal emulators for details) that your
terminal or terminal PC uses. The additional questions depend on the emulation selected. The
possible emulations are:
1 = GenTerm in WT emulation mode. This emulation is fully tailored to be used with
MicroBeta, so the only things the user has to specify are (see example 9):
Example 9
System>M
Terminal emulation:
1)GenTerm 2)UltroTerm 3)MultiCalc
4)VT52
5)VT100
6)MBW
2->1
Number of lines on screen
24->
Print through terminal (Y/N)
N->
Do you accept the new terminal setting? (Y/N)->N

- the number of lines on the screen.

- whether the printer should be connected to the terminal PC or not (see 2J.30.10.5).
2 = UltroTerm (see example 10). Select this if you are using the UltroTerm terminal emulator.
UltroTerm V2.0 or later is required.
Example 10
System>M
Terminal emulation:
1)GenTerm
2)UltroTerm
3)MultiCalc
4)VT52
5)VT100
6)MBW
2->
Number of lines on screen
23->
Print through terminal (Y/N)
N->
Do you accept the new terminal setting? (Y/N)->Y
Prepare your terminal ready for emulation change.

238

2J.30 System

Press any key when ready.

3= MultiCalc VT52. Extensions to enable result file saving to PC disk when using MultiCalc.
4 = VT52 (see example 11). Select this if you are using an emulator that can provide only this
emulation.
5 = VT100. This emulation is quite popular in commercial terminal emulation programs. If
your terminal emulator implements both VT52 and VT100, select VT100 because it supports a
better range of MicroBeta features i.e. counting results displaying (screen attributes, result
window scrolling etc.).
6 = MBW (MicroBeta Windows workstation VT52). If this is selected then results sorting and
live data automatic sending must be specified. The workstation program sets the terminal and
other settings automatically.
Additional questions:
2J.30.10.1 Number of lines on screen
Possible values from 15 to 25, the default value is 24 lines. Some terminal emulator programs
can scroll the whole screen if something is written at the lower right corner of screen (e.g.
column 80 on line 24) which mixes the status display. If you encounter this problem give here
a smaller value (e.g. 23 lines). You can select a smaller value also if your terminal PC cannot
display 24 lines (e.g. some portable PCs can display 16 lines).
2J.30.10.2 Block graphics
For VT52 and VT100, default Y=Yes. If your terminal shows some strange characters around
the status display and it seems that it cannot display block graphic characters, select N=No.
2J.30.10.3 Send result files to terminal
For VT52 and VT100. The method of sending result files to the terminal PC depends on the
terminal emulator software. Often a terminal emulator can capture data between a start string
and a stop string. If your terminal emulator is capable of doing this then answer Y=Yes. Then
MicroBeta stores a result file at first on the protocol diskette and sends it to the terminal as one
block as:
Start of assay:XX
... ... Result data
...
End of assay

239

2J.30 System

where xx is the protocol number.

If your terminal emulator cannot capture data then answer N=No. Then the result files are
stored on the protocol diskette of MicroBeta but not transferred to the terminal PC.
Note 1: Result file will be output if protocol line 36 FILE OUTPUT is set to other than 1 (No
output).
Note 2: See chapter Results for details.
2J.30.10.4 Automatic result file deletion
For VT52 and VT100, default Y=Yes. The capacity of the protocol diskette is limited (300KB
- 1.3MB depending on the number of protocols) and it is capable of storing the results from at
least 20 cassettes if the output selection for FILE OUTPUT does not specify too many different
printout fields. If this selection is set to Y=Yes then the result files on protocol diskette will be
deleted automatically when starting counting and after sending a result file to the terminal PC
is complete. If you want to ensure that your data has been transferred to another diskette or to
some PC then answer N=No. In that case after starting counting the program confirms deletion
before operation by asking:
Delete result files saved on protocol disk?

and you can answer N=No to reject deletion or Y=Yes to accept deletion.
2J.30.10.5 Print through terminal
Select here where to connect the printer. Answer: N=No if you do not need to connect the
printer to the terminal PC. Connect instead the printer to serial port 3 of MicroBeta with serial
cable 1221-502.
Y=Yes to connect the printer to the terminal PC. In this case connect the printer to the parallel
port of the terminal PC with a Centronics cable 1221-122.
2J.30.10.6 Exit terminal setting
To exit terminal setting press slash / and Enter on any line. Otherwise, after you have answered
all the questions described above, the program confirms that you accept the change. If not,
press N, or else press Y which causes the following message to appear:
Prepare your terminal ready for emulation change. Press any key when ready.

Now you can make the preparations possibly required for the emulation change (e.g. to switch
from GenTerm to UltroTerm). When ready press any key. Then if the change succeeded the
program returns to the System state. However if the message Terminal type check failed!
240

2J.30 System

appears (see example 11) it means that MicroBeta has found that the terminal program does not
use the same emulation as was selected (MicroBeta sends an identity inquiry to the terminal
and the answer sent by the terminal emulator software is different for each emulation GenTerm, UltroTerm, VT52, VT100). Answer N if you want to cancel the new terminal
emulation. However if you know that your terminal has the correct emulation or you want to
switch to the new emulation later you can answer Y.
Example 11
System>M
Terminal emulation:
1)GenTerm 2)UltroTerm 3)MultiCalc
4)VT52
5)VT100
6)MBW
2->4
Number of lines on screen
24->
Block graphics (Y/N)
Y->
Send results file to terminal (Y/N)
N->
Automatic result file deletion (Y/N)
Y->
Print through terminal (Y/N)
N->
Do you accept new terminal setting? (Y/N)
->Y
Prepare your terminal for emulation change.
Press any key when ready
Terminal type check failed!
Do you accept the new terminal setting? (Y/N)->N

2J.30.11 P(a)sswords
Press A to print protocol passwords, entered on line 45 in the protocol editor (see chapter 2J.25
Protocols). This is possible only if no System password is in use or in Setup mode (see
2.30.15).
Example 12
System>A
Printing protocol passwords, press / to exit

2J.30.12 Customi(z)ing
Press Z to set environment strings to customize the program. The environment string name and
value must be given. For a list of possible strings, see (I)nfo/Customi(z)ing. Giving D as the
value sets the default value for the string.
Example 13
System>Z
Customized: MENU = N
Environment string name

-> MENU

241

2J.30 System

MENU
Environment string name

-> D
-> /

2J.30.13 MS-D(O)S
Press O to get to the MicroBeta MS-DOS. You must confirm this twice as example 14 shows,
after which the text MicroBeta A:\> appears:
Example 14
System>O
Exit to MS-DOS? (Y/N) ->Y
Are you sure? (Y/N) ->Y
Type MB to start the program again
MicroBeta A:\>MB
(H)elp
(I)nfo
(C)ount
(P)rotocols

(S)ystem

Ready>

Type MB and press Enter to get back to the MicroBeta program.

Note: Avoid going to MS-DOS in routine work. The main reasons to go there are to make a
backup copy of the protocol or program diskette.

2J.30.14 (I)njector
Press I to set injector parameters for MicroBeta JET.

242

2J.30 System

Example 15
System>I
Mask adapter (Y/N)
Y->
Injector module (Y/N)
Y->
Raise detector (Y/N)
Y->
Multichannel pump (Y/N)
Y->
Number of channels
6->
Number of injectors
2->
Injector 1:
Syringe volume: 1)100uL 2)250uL 3)500uL
3->
Aspirating tube I.D. ["]
0.053->
Aspirating tube length ["]
27.0->
Dispensing tube I.D. ["]
0.030->
Dispensing offset [uL]
0->
Injector2:
Syringe volume: 1)100uL 2)250uL 3)500uL
3->
Aspirating tube I.D. ["]
0.053->
Aspirating tube length ["]
27.0->
Dispensing tube I.D. ["]
0.030->
Dispensing offset [uL]
0->
Dispense fast command string S10L7IAxOv1000V4000c2700L20A0R ->
Dispense slow command string S10L7IAxOv1000V2000c2700L20A0R ->
Dispense custom command string S10L7IAxv1000V3000c2700L20A0R ->
Wash command string
S10IAxV3800OA0R ->
Tip init command string
S10L7OM200A0A24IR ->
Prime command string
S10IAxOV3800A0R ->
Suck-back command string
S10OAxIV3800A0R ->
Initialize command string
Z6R ->
Stop command string
T ->
Dispensing volume max [uL]
250 ->
Store data? (Y/N)
->

2J.30.14.1 Mask adapter.

This is the plate that fits between the detectors and the microtitration plate. The number of
holes in the plate correspond to the number of detectors. This parameter allows you to select if
the mask adapter is to be used. It must be used if you want to use the injector system since the
injector tubing is fitted into the mask adapter.
2J.30.14.2 Injector module
This parameter must be Y if you want to use injection.
2J.30.14.3 Raise detector
This parameter must be Y if you want to use injection because the upper detector must be
raised to make space for the mask adaptor and injector(s).

243

2J.30 System

2J.30.14.4 Multichannel pump

There can be from 1 to 6 channels for the pump. If the pump is a single channel one, i.e. for a
single detector MicroBeta JET then select N here. Otherwise select Y for a 2, 3 or 6 channel
pump.
2J.30.14.5 Number of channels
Give here the number of channels 1, 2, 3 or 6.
2J.30.14.6 Number of injectors
A single channel system can have up to four injectors, whereas a multiple channel system can
have a maximum of two injectors. Give the number for this parameter.
2J.30.14.7 Injector 1
As many injectors will appear as there are selected. Each injector has its own set of paramters
as follows.
2J.30.14.8 Syringe volume
This is the amount of liquid a syringe takes in during one stroke. There are three possibilities
100, 250 or 500 L. Select the one you want.
2J.30.14.9 Aspirating tube I.D.
The internal diameter of the tubing from the liquid reservoir to the syringe. The units are
inches.
2J.30.14.10 Aspirating tube length
The length of the tubing from the liquid reservoir to the syringe. The units are inches.
2J.30.14.11 Dispensing tube I.D.
The internal diameter of the tubing from the syringe passing through the injector assembly to
above the well.
2J.30.14.12 Dispensing offset
This is the offset volume for injector modules and it is set in steps of 1 L. It is a volume that
effects the injector dispensing volume. If the volume is selected from the protocol as 10 L and
the offset is 0, then the dispensed volume is 10 L. If the offset is 2 L then when the protocol
selection is 10 L the real dispensed volume is 12 L. So if in the injector there is for some
reason a systematic error in the dispensing volume it can be corrected with the offset.

244

2J.30 System

The following parameters are common for all channels and should not be changed by the user
without contact with a service person.
2J.30.14.13 Dispense fast command string
This defines how fast the dispensing occurs for fast dispensing.
2J.30.14.14 Dispense slow command string
This defines how fast the dispensing occurs for slow dispensing.
2J.30.14.15 Dispense custom command string
This defines how fast the dispensing occurs for the custom dispensing. This can be set to the
speed you require if neither slow nor fast speeds are acceptable.
2J.30.14.16 Wash command string
This defines how the wash procedure occurs. Wash is used to clean the tubing after use.
2J.30.14.17 Tip init command string
This ensures that there are no drops on the end of the tubing.
2J.30.14.18 Prime command string
This fills the tubing with liquid before operation can begin.
2J.30.14.19 Suck-back command string
This command causes the liquid in the tubing to be returned to the reservoir.
2J.30.14.20 Initialize command string
This prepares the system for use and is the first operation before any other injection operation.
2J.30.14.21 Stop command string
The is the command to stop operation.
2J.30.14.22 Dispensing volume max.
This is the maximum volume that may be in a well. It is the sum of the sample volume and the
dispensed volume. It ensures that the well is not overfilled.

2J.30.15 (Q)uit
Press Q to quit from the System state.

245

2J.30 System

2J.30.16 Se(t)up mode

If a system password is used then the system mode menu is quite short (see example 16).
Example 16
(H)elp
(E)rrorbeep
(C)lock

(Q)uit
(S)tatus display
Se(t)up mode

MS-D(O)S
(V)ersion

Ready>T
Password ->
(H)elp
(E)rrorbeep
(C)lock
(D)ata drive
Customi(z)ing

(Q)uit
(S)tatus display
(R)S-232C
Ter(m)inal
(I)jector module

MS-D(O)S
(V)ersion
(P)rinter
P(a)sswords

System>

All the functions that can affect routine use are hidden. The person responsible for the
MicroBeta system can make the hidden functions visible if he or she knows the password.
Press T to get into the setup mode. The program then asks for the password. Type the password
and press Enter. The characters you type are not displayed on the screen. If you enter a wrong
password the program displays the message:
Incorrect password

Otherwise the program shows the full System menu.

To set a system password, exit from the MicroBeta program to MS-DOS as described earlier.
After the text MicroBeta A:\> appears, type PASSWORD xxxx and press Enter (xxxx means the
new password). If you forget the password then set a new one.

246

Part 3 General description

247

248

3.1 Instrument description

3.1 Instrument description

3.1.1 Introduction
This section describes the hardware features of 1450 MicroBeta TriLux and MicroBeta JET.

3.1.2 Combined liquid scintillation and luminescence counter

With MicroBeta TriLux you can count beta or gamma emitting samples or glow luminescence
samples. With MicroBeta JET you can in addition count flash luminescence samples in which
reagent injection is required. You get the luxury of several powerful counting techniques
combined in one counter.

3.1.3 Microtitration plate based operation

In MicroBeta samples are counted in a microtitration plate. Before counting, the instrument
itself determines which type of plate is being used.
The plate is placed in a special cassette for counting which you can identify with barcoded
labels. Load the cassette into the cassette rack behind the door with the plate upwards and the
ID labels towards you, then close the door.

3.1.4 Cassette rack and transportation system

The cassette rack is located inside the door and consists of 16 (32) shelves. These are
numbered 1 to 16 (32) from the bottom upwards. The maximum capacity is 1536 (3072)
samples when using 96-well plates and 384 (768) samples when using 24-well plates and 6144
(12288) samples when using 384-well plates.
The cassette rack is in its middle position when being loaded. A motor moves the rack up or
down one step at a time. Sensors are used for identifying shelf numbers and for checking if
there are cassettes on the shelves and whether they are 24, 96 or 384-well type. A sensor will
cut off the high voltage if the door is opened during counting in order to protect the detectors.
One cassette at a time is moved to the transportation system. This is an X-Y transfer
mechanism which first moves it to the ID reader which reads the barcodes on the cassettes.
Then it is moved to the first counting position. After a cassette has been counted, it is moved
back to the rack. The elevator moves the cassette rack down one position and the next cassette
is moved to the transportation system.

249

3.1 Instrument description

3.1.5 Coding
Code labels can be used to identify which protocol (set of counting parameters) is to be used
for counting the samples in a cassette. These identification labels are printed with barcodes,
and the reader is situated in front of the detector block. There are identification labels for
identifying the protocol and the cassette number. There are also three special function codes:
STOP, NORM and STD. When a cassette carries a STOP code, it is counted normally, and
after it the counting stops. NORM and STD show that the cassette carries normalization or
standardization samples respectively. See chapter 2.4 Coding the cassette for more details. You
can also identify the plate itself with a barcode, see section 2.4.6 Plate ID reader.

3.1.6 Detectors
One, two, three, six or in the case of MicroBeta TriLux twelve pairs of photomultiplier tubes
view the samples in the measuring position. The detectors are placed in a 1x1, 2x1, 1x3, 2x3 or
3x4 formation respectively, see chapter 2.9 Detectors for pictures of this formation. The tubes
of one pair are situated on opposite sides of the counting block, one above and one below the
block. Each pair operates in coincidence mode which greatly reduces single photon
background events. The out-of-coincidence signal is also measured simultaneously.
When a cassette moves into the counting position the labels on the cassette are read by the
barcode reader. Then 1, 2, 3, 6 or 12 samples on the cassette are counted simultaneously
according to the protocol specified by the label on the cassette.
Due to the relatively small sample volume there is such a low background count that no extra
lead shielding is needed, thus reducing the weight of the instrument.
To avoid ambient light affecting the photomultiplier tubes, the inside of the instrument is
painted black and the door has a light seal along its edge.
To protect the detectors, the high voltage of the photomultiplier tubes is cut off when the door
is open. A warning message is displayed if counting is attempted while the door is not properly
closed. See chapter 2.16 Interrupt.

3.1.7 Glow luminescence assays

With MicroBeta TriLux, luminescence assays designed to be completed on a solid support e.g.
nylon membrane, glass fibre or coated plate, and in solution can be directly quantified.
Due to the high signal, luminescence reactions can be measured using single PMTs.

250

3.1 Instrument description

Traditional microtitration plates are totally opaque (black or white). These plates are counted
with the upper tube and a 1450-105 cassette is used. The filters are counted with the upper or
lower tube and a 1450-104 filter cassette is used.
A special feature when using MicroBeta as a luminometer is the 1450-466 coloured sealing
tape. The saturation of PMTs is prevented and crosstalk between adjacent positions is
minimized by cutting the intensity of luminescence light with the 1450-466 coloured sealing
tape.
The luminescence units used are LCPS: luminescence counts per second i.e. corrected CPSvalues/100.

3.1.8 MicroBeta JET features

In addition to the MicroBeta TriLux features described above, MicroBeta JET makes it
possible to measure flash luminescence, dual luminescence and do kinetic studies because it
allows reagent addition and mixing by dispensing.
MicroBeta JET can be purchased without the injector assembly or injector modules and these
can be added later in the field by the user when required. Also additional injector modules can
be added.
An injector module is a pump with 1, 2, 3 or 6 syringes each feeding tubing that goes to the
mask adapter which positions the tips of the tubes above the well(s). Each syringe and hence
piece of tubing constitutes one channel. A single detector instrument can have up to four
modules each with one channel. A multiple detector instrument can have up to two modules,
each with as many channels as there are detectors in the instrument i.e. 2, 3 or 6.
Each module and channel can be switched on or off independently.
The use of at least two injectors for the same well means that dual luminescence assays can be
performed.
If reagent injection is selected in any particular counting protocol, the mask adapter (injector
assembly) is driven into position above the microplate. The injector tips which are embedded
in the mask adapter are automatically positioned over the wells to allow injection under
optimum conditions. The light resulting from the reaction passes through the fibre optic
coupling in the injector assembly to the upper PMT.

251

3.1 Instrument description

The mask adapter can also be used without injection simply for masking. It reduces the signal
to the upper detector by a factor of ten which can be helpful in cases of luminescence with very
high signal.

3.1.9 Robotic loading interface

The robotic loading interface is a special door/adapter system that brings a cassette outside so
that a robotic arm can change the plate. In this case only one shelf is used.
Note: the mask adapter has to be moved manually into the counting chamber before starting
counting.

3.1.10 Microcomputer control

Operation of the instrument and calculation of the results is performed by a combination of a
terminal PC running a Wallac terminal interface program or the MultiCalc immunoassay
program, and the instrument itself.
The built-in computer is composed of a 16-bit Intel 80186 microprocessor with 1 MB RAM
and 64 kB ROM. There are two microfloppy 3.5 disk drives; each disk has a capacity of 1.44
MB. The disks loaded onto these drives contain the program and the protocols.
The Terminal PC is a personal computer that is used to run the terminal emulation program or
MultiCalc and can be used to collect counting data for further evaluation.
Output possibilities can be selected by means of the programmable RS-232C interface with
three output ports. The uppermost port sends the output to the terminal, the second to an
external computer and the third to a printer or in the case of MicroBeta JET to the injector
modules.
Should any fault occur, the built-in service test routines allow the service engineer to quickly
diagnose the cause of the fault and, due to the flexible arrangement of the electronics, easily
change any components where necessary.

252

3.2 Routine maintenance

3.2 Routine maintenance

3.2.1 Cleaning the instrument
The cassette rack should be kept clean. The cassettes should be kept clean in order to prevent
dirt from coming between the detector blocks, to minimise the background and maximise the
light collection efficiency.
In MicroBeta JET the optical fibres of the mask adapter (injector assembly) must be kept clean.
If necessary, wipe them with a moist cloth.
Tubing should be washed with water using the Wash command.
Note: refer also to the reagent manufacturer's instructions to see if there is additional
information on cleaning tubing.

3.2.2 ID label
Check the labels on the ID support plates. Those in bad condition should be replaced by new
ones to guarantee correct reading of the labels. When fixing ID labels on the support plate,
ensure that the area where the label is to be fixed is clean, e.g. that there is no perspiration from
your fingers on it.

3.2.3 Power supply fan

Check that the cooling fan in the power supply unit is working by listening for its sound.

3.2.4 Checking the instrument

To check the instrument, it is recommended that at least once a month you run counting
protocol number 99, the GLP (Good Laboratory Practice) protocol. Use this protocol to count
the normalization sample plate supplied with the instrument. The background, 3H and 14C
samples are counted with all detectors so background level, CPM values and SQP(I) values can
easily be compared.
Recommended limits:
3
14
H
C
Eff%
Bkg
Eff%
Bkg
>45%
max 50 cpm
>91
max 60 cpm
If the values obtained by running the GLP protocol are outside the recommended limits, please
contact your service support person.

253

3.2 Routine maintenance

254

3.3 Calculation methods

3.3 Calculation methods

3.3.1 Count rate
Count rate i in counts per minute (headings CPM, CPM1, CPM2, CPM3) is calculated using
the equation:
i =60*c/t

(1)

where c is counts in the counting window and t is dead time corrected counting time (heading
CTIME) in seconds.
In ParaLux high efficiency mode:
CPM1 = CPM_C + CPM_NC
where CPM_C is normal coincidence count rate and CPM_NC is non-coincidence count rate
between discriminator channel and 1024.
Count rate -sigma percentage error (headings CPM%, CPM1%, CPM2%, CPM3%) is
calculated using the equation:
*i/i = * 100 / c

(2)

Above and in what follows amu can be either 1.0 or 2.0 .

3.3.2 Normalization
3.3.2.1 Detector efficiency normalization
Detector efficiency normalization corrects for differences in detector responses. Efficiencies
and efficiency errors of each detector are calculated and stored for count rate correction.
Background corrected count rate r is calculated using the equation
r=i-b

(3)

where i is from equation (1) and b is the normalization background count rate for the same
detector and counting window. ParaLux high efficiency backgrounds are subtracted first.
Efficiencies e(mn) are calculated using the equation

255

3.3 Calculation methods

e(mn) = r(mn) /j

(4)

where r(mn) is isotope n background and half-life corrected count rate in window m and j is the
given normalization activity or the maximum background corrected count rate of isotope n in
window 1 (single label) or in window 5-1024 (dual label) or the average count rate
(luminescence).
Efficiency percentage error is calculated using the equation
e(mn)/e(mn) =
{[i(mn)/r(mn)]2 +[b(mn)/r(mn)]2 +[j/j]2 }

(5)

If j is the given activity then j=0.

3.3.3 Corrected count rate

Corrected count rate (headings CCPM, CCPM1, CCPM2) is calculated in three steps in the
following order:
- Background correction
- Detector efficiency correction or crosstalk correction
- Half-life correction
The first two corrections also affect the theoretical error (headings CCPM%, CCPM1%,
CCPM2%) of the corrected count rate.
3.3.3.1 Background correction
If background sample is specified and measured or background is given in the protocol then
count rate i is corrected for background using equation (3).
r=i-b
ParaLux high efficiency backgrounds are subtracted first.
If background sample is measured then the count rate -sigma error becomes:

256

3.3 Calculation methods

*r /r = [ (i)2 + (b)2] / r

(6)

3.3.3.2 Detector efficiency correction

In single label counting detector efficiency correction for count rate is calculated using the
equation
R=r/e

(7)

where r is from equation (3) and e = e(11) from equation (4) for the same detector as in
normalization.
Efficiency correction affects also the corrected count rate -sigma error,
*R /R = [ (r/r)2 + (e/e)2]

(8)

where e/e = e(11)/e(11) from equation (5) and r/r is from equation (6).
In dual label counting the efficiency correction has the form
R(m) = [e(nn)*r(m) - e(mn)*r(n)] / d

(9)

where
d = e(mm)*e(nn) - e(nm)*e(mn)

(10)

and e(mn) stands for isotope n efficiency in window m.

Corrected count rate -sigma percentage error in the dual label case is calculated using the
equation
* R(m)/R(m) =
*100[[r(m)2 * e(nm)2 + e(nm)2 * r(m)2 + r(n)2 * e(mm)2 +e(mm)2 *r(n)2]/p(m)2+Q/d2]
(11)
where r(m) is from equation (3), r(m) is from equation (6), d is from equation (10),
p(m) = e(nn) * r(m) - e(nm) * r(n)

(12)

257

3.3 Calculation methods

and
Q = e(mn)2 * e(nm)2 + e(nm)2 * e(mn)2 + e(nn)2 * e(mm)2 + e(mm)2 * e(nn)2

(13)

3.3.3.3 Half-life correction

Half-life corrected count rate I is calculated using the equation
I = [[(t/)*ln(2)]/{exp[-ln(2)*(T-t)/]-exp[-ln(2)*(T/)]}]*R

(14)

where t is counting time, T is elapsed time from zero time to the end of counting, is the halflife of the isotope and R is the background and detector efficiency corrected count rate. Both
single and dual label corrections have the same form. Half-life correction has no effect on the
corrected count rate errors.

3.3.4 Spectrum indexes

3.3.4.1 Isotope spectrum end point
Isotope spectrum end point (heading ISEP) is a channel at the end of the spectrum.
3.3.4.2 Spectrum quench parameter
Spectrum quench parameter of the isotope spectrum (heading SQP(I)) is the spectrum mean
pulse height and is calculated using the equation
q = [i * c(i)] / c(i)

(15)

where i = window 1 low .. Max (window 1 high, window 2 high, window 3 high) is the channel
number and c(i) is the number of counts in channel i. The SQP(I) counting window is truncated
so that a 10 counts range is removed from both ends.
The - sigma percentage error of SQP(I) is given by:
*q/q = *100 ({[i2 c(i)]/c(i) - q2}/c(i))/q

(16)

3.3.4.3 Asymmetric quench parameter

Asymmetric quench parameter (heading AQP(I)) is a measure of the isotope spectrum for an
asymmetric sample and varies as the quench changes. It establishes a relation between quench
and efficiency.

258

3.3 Calculation methods

3.3.5. Standardization
3.3.5.1 Activity standardization
Standardization corrects for differences in sample quench levels and in detector responses.
All standard samples are counted with detector 1. Only the unquenched standard N1 and the
fully quenched standard N2 are counted with all detectors. The SQP(I)s or AQP(I)s,
efficiencies and weights of standard samples are calculated and stored for activity calculations.
The efficiency in a counting window is
E = r/a

(17)

where r is background and half-life corrected count rate and a is the given activity of the
standard sample.
Standardization weight is given by
w = [(k*q)2 + (r/a)2]

(18)

where k is the slope of the standard curve at q, q is from equation (16) and r is from equation
(6).

3.3.6. Activity
3.3.6.1 Using activity standardization
Sample activity in disintegrations per minute (headings DPM and DPM1) is
A = r/E

(19)

where r is background corrected count rate (3) and E is counting efficiency (17) at SQP(I) or
AQP(I). Activity is half-life corrected using equation (14).
Efficiency E is obtained from the standardization efficiency curve of detector 1 using the
measured SQP(I) or AQP(I) of detector i.
The measured SQP(I) or AQP(I), q(i), is first corrected to correspond to detector 1,
q = q(1,N2) + [{q(1,N1)-q(1,N2)}/{q(i,N1)-q(i,N2)}]*[q(i)-q(i,N2)]

259

(20)

3.3 Calculation methods

where the first index stands for the detector number and the second index stands for the
standard sample type.
If the measured SQP(I) or AQP(I) is not between the standardization SQP(I) or AQP(I) limits,
then linear extrapolation with the two first or last points is used (or spline with the two last
points if smoothing spline is selected). If the DPM out-of-range monitor (heading DPMM) is
selected then OUT is printed to warn the user.
The interpolated efficiency is corrected to correspond to detector i using the equation
E = E(i,N2) + [{E(i,N1)-E(i,N2)}/{E(1,N1)-E(1,N2)}]*[E(1)-E(1,N2]

(21)

Activity -sigma percentage error (headings DPM% and DPM1%) is given by

*A/A = [(r/r)2 + (E/E)2]

(22)

where r/r is count rate error (6) and efficiency error comes from the equation
E = [(k*q)2 + w2].

(23)

Here k is the slope of the standard curve and w is the standardization weight at q.

3.3.7 Chemiluminescence correction

Chemiluminescence correction is done by subtracting the chemiluminescence counts from
window 1,
c(corr) = c - c(chem).

(24)

Chemiluminescence counts are calculated using the equation:

c(chem) = 7.4E-9 * c(l) * c(r)

(25)

where c(l) and c(r) are the lower and upper detector counts respectively.
The percentage of chemiluminescence in window 1 (heading CLM%) is given by
l = 100 * [c - c(corr)]/c

(26)
260

3.3 Calculation methods

If CPM1 < 10 then chemiluminescence percentage is always zero.

3.3.8 Statistics monitor

If statistics monitoring is selected (heading STM) then the counting time is divided into ten
parts and the samples are recounted if bad statistics is detected.
The counting time must be at least ten seconds for the statistics monitor to function. E.g. 53
seconds is divided into 10 times 5 seconds and the last 3 seconds is not monitored. ., REC1,
REC2 or ERR is printed for samples with no recounting, one recounting, two recountings or
two recountings and still bad statistics. Using the next sample function disables the statistics
monitor. If counting ends because of precision then the statistics monitor functions only if at
least nine parts have already been counted.
Statistics is defined as bad if the chi-square value
2 = O2/T2 = [{i2 - n * i(mean)2}/{n-1}] /[(1/n)(i/t)]>2.4

(27)

Here O is the observed deviation and T is the theoretical deviation. The count rate during time
part t is i and the number of parts n = 10.

3.3.9 Statistical calculations

3.3.9.1 Mean
The mean value of corrected count rate or activity A from n repeats or replicates (headings
MEAN, MEAN1, MEAN2) is calculated using the equation
M = A / n

(28)

where the sum is over the repeat or replicate number.

3.3.9.2 Theoretical standard error of mean
The theoretical percentage standard error T of the mean M (headings THEOR.ERROR(%),
TSEM1%, TSEM2%) is calculated using the equation
T = *100 [(A)2] / (n * M)

(29)

where A is the corrected count rate or activity -sigma error.

261

3.3 Calculation methods

3.3.9.3 Standard error of mean

The observed standard error O of the mean M in percentages (headings OBS.ERROR(%),
SEM1%, SEM2%) is calculated using the equation:
O = *100{(M-A)2/[n(n-1)]}/M

(30)

= *100{[A2 - (A)2/n]/[n(n-1)]}/M
3.3.9.4 Theoretical coefficient of variation
The theoretical coefficient of variation CV% of the mean M in percentages (headings TCV1%,
TCV2%) is calculated using the equation:
TCV% = *100 {[ (A)2]/n}/M

(31)

3.3.9.5 Coefficient of variation

The observed coefficient of variation CV% of the mean M in percentage (headings CV1%,
CV2%) is calculated using the equation:
CV% = *100{(M-A)2/(n-1)}/M

(32)

= *100{[A2 - (A)2/n]/(n-1)}/M
3.3.9.6 Theoretical standard deviation
The theoretical standard deviation TSD (headings TSD1 and TSD2) is calculated using the
equation:
TSD = * [(A)2/n]

(33)

3.3.9.7 Observed standard deviation

The observed standard deviation SD (headings SD1 and SD2) is calculated using the equation:
SD = *{[A2 - (A)2/n]/(n-1)}

(34)

3.3.9.6 Chi-square test

The reduced chi-square value 2 (headings CHI-SQUARED, CHISQ1, CHISQ2) is defined as
2 = O2 / T2

(35)

262

3.3 Calculation methods

where the observed error O is from equation (30) and the theoretical error T is from equation
(29).
How much 2 deviates from unity is a direct measure of the extent to which O deviates from T.
With a limited number of measurements we would not expect these two errors to be precisely
the same. Thus a more quantitative test is required to determine whether the observed
difference between these two deviations is really significant. For this purpose we can apply the
chi-square test.
From the 2-value we can calculate the probability (headings PROBABILITY(%), P1, P2) that
a random sample from a correct distribution would have a larger value of 2 than the observed
2-value.

263

3.3 Calculation methods

264

3.4 Specifications

3.4 Specifications
3.4.1 Physical dimensions
Width
Height
Depth
Weight

TriLux
430 mm
610 mm
645 mm
655 mm 12 det.
85 kg
90 kg 12 det.

JET
700 mm
630 mm
660 mm
90 kg

3.4.2 Power
Mains voltage selectable 100, 115, 120, 220, 240 V, +/- 10 %, 50/60 Hz, power consumption
360 VA.
Injector module: 24 V +/- 10 %, max 2A.

3.4.3 External power supply

Power failure recovery and protection can be obtained with an external UPS back-up power
supply provided. This can be ordered from PerkinElmer Life and Analytical Sciences.

3.4.4 Connections/input/output
Serial ASCII interface RS-232 C. Connector No.1 for terminal computer, No.2 for mainframe
or data net and No.3 for printer.

3.4.5 Operating conditions

Temperature +15 to +35oC.
Humidity max. 10 - 80 %.
Pollution degree 2.
Installation category II.
Altitude 2000 m.

265

3.4 Specifications

3.4.6 Sample capacity

Sample capacity using 96-well sample plates is 16(32)*96 = 1536(3072) samples, using 24well sample plates 16(32)*24 = 384(768) samples and using 384-well sample plates
16(32)*384 = 6144(12288). Any combination of cassettes either for 96 or 24-well sample
plates or 96 or 384-well sample plates is possible.

3.4.7 Sample cassette changer

A z-direction moving cassette rack with shelves for 16 or 32 cassettes. This rack functions as a
random access stack which allows access to any cassette in the rack. The cassette rack is driven
by a stepper motor and its movement is controlled by electro-optical sensors. A transportation
system moving cassettes in the x-y-direction selects one cassette from the cassette rack and
takes it into the counting position and then returns the cassette to the cassette rack. This
transportation system is driven by two stepper motors and is controlled by electro-optical
sensors.

3.4.8 Mask adapter (Injector assembly)

A transportation system for moving the mask adpater in the x-direction from the maintenance
tray into the counting chamber under the upper detector block. This transportation system is
driven by one stepper motor and is controlled by electro-optical sensors. The mask adapter
moves in the z-direction with the upper detector block.

3.4.9 Detector assembly

1, 2, 3, 6 or 12 detectors (each consisting of a pair of 19 mm PM tubes working in
coincidence), are mounted in two detector assemblies, the upper detector assembly moving in
the z-direction. PM tubes are Hamamatsu R1166 or equivalent. Coincidence resolution time is
100 ns. One to six detector assemblies are provided with circular apertures for 96 and 24-well
sample plates. The 12-detector assembly is provided with circular apertures for 96-well sample
plates and square apertures for 384-well sample plates. The apertures are automatically
changed to correspond to the plate type.

3.4.10 Cassettes
Cassettes are made of special-grade polycarbonate plastic containing a high amount of white
pigment to produce a very high degree of reflectivity (about 90 %) while eliminating optical
crosstalk between wells. Measurement is made through the cassettes (pat. pend.).

266

3.4 Specifications

3.4.10.1 96-pos. cassette 1450-101

This has 96 sample holes with a diameter of 7.8 mm. Well separation is 9 mm. The cassette is
used when counting 1450-401 (or equivalent) 96-well sample plates.
3.4.10.2 24-pos. cassette 1450-102
This has 24 sample holes with a diameter of 13.2 mm. Well separation is 18 mm. This cassette
is used when counting 1450-402 (or equivalent) 24-well sample plates.
3.4.10.3 96-pos. cassette 1450-103
This has 96 sample holes. The cassette is used when counting 1450-407 plates.
3.4.10.4 96-pos. cassette 1450-104
This is a two piece cassette with 96 sample holes. This cassette is used when counting
filtermats. The filtermat in a sample bag is placed between the base plate and the cover plate.
3.4.10.5 96-pos. cassette 1450-105
This has 96 sample holes. This cassette is used when counting 1450-410, 1450-405, 1450-419
or equivalent sample plates.
3.4.10.6 96-pos. cassette 1450-106
This has 96 sample holes. This cassette is used when counting e.g. Millipore MultiScreen
Filtration plates or equivalent sample plates.
3.4.10.7 24-pos. cassette 1450-107
This is a two piece cassette with 24 sample holes. The cassette is used when counting Skatron
filtermats cut into four pieces. The filtermat in a sample bag is placed between the two plates of
the cassette.
3.4.10.8 24-pos. cassette 1450-110
This is an open-based cassette. The cassette is used when counting 24 well culture plates, e.g.
Costar plates (or equivalent).
3.4.10.9 24-pos. cassette 1450-116
This is a two piece cassette with 24 sample holes. This cassette is used when counting
filtermats. The filtermat in a sample bag is placed between the base plate and the cover plate.
3.4.10.10 24-pos. cassette 1450-117
This is a two piece cassette with 24 sample holes. This cassette is used when counting 4 mL
sample vials. The vials are placed between the base plate and the cover plate.

267

3.4 Specifications

3.4.10.11 96-pos. cassette 1450-118

This is a two piece cassette with 96 sample holes. This cassette is used when counting e.g. 32Plabelled dot blot samples from filtermats such as 1450-423 Nylon membrane. No scintillator
needs to be added to the filtermat allowing reprobing assays. The filtermat in a sample bag is
placed between the base plate and the cover plate having solid scintillator in sample positions.
3.4.10.12 384-pos. cassette 1450-130
This has 384 sample holes. the cassette is used when counting 384-well sample plates.
All the cassettes are made of white plastic with an optical surface in the sample holes.

3.4.11 Sample identification system

Each cassette can be provided with a 1450-451 ID Support Plate for 1450-452 ID Labels. An
ID Support Plate has marked areas for ASSAY, PROTOCOL and CASSETTE numbers (0 99) and FUNCTION codes (CPM-normalization, DPM-standardization, stop). This provides
the information needed for Good Laboratory Practice.
There is also an optional Plate ID reader.

3.4.12 Electronics
3.4.12.1 Environmental
Electromagnetic susceptibility according to: EN50082-1.
Electromagnetic emissions according to: EN500081-1.
The following interconnecting cables should be used to comply with these standards:
Instrument to PC: 1221-503
Instrument to printer: 1221-502
3.4.12.2 Hardware
16-bit microprocessor controlled counting and data reduction. Memory configuration 64 kbyte
ROM and 1 Mbyte RAM. Two 3.5 inch 1.44 Mbyte disk drives: drive A for program disk and
drive B for protocol disk.
Logarithmic A/D converter energy range 1 - 2000 keV. There is a dual 1024-channel
multichannel analyzer for each detector. Built-in dead time correction.

3.4.13 Performance
(100 ns coincidence time)
268

3.4 Specifications

3.4.13.1 Background
(at the factory, Turku, Finland):
With empty 1450-514 plate:
<15 CPM in wide 3H window
<17 CPM in wide 14C window.
Unquenched sample with a volume of 150 microlitres high flash point cocktail unpurged in the
1450-401 96-well Sample Plate: typically 6 CPM in wide 3H window typically 9 CPM in wide
14
C window.
Unquenched sample with a volume of 750 microlitres high flash point cocktail unpurged in the
1450-402 24-well Sample Plate:
typically 24 CPM in wide 3H window
typically 37 CPM in wide 14C window.
3.4.13.2 Efficiency
1450-471 Normalization Standard
3
H > 45 %
14
C > 90 %
Unquenched sample with a volume of 150 microlitres high flash point cocktail unpurged in the
1450-401 96-well Sample Plate:
3
H: typically 57 %
14
C: typically 94 %
Unquenched sample with a volume of 750 microlitres high flash point cocktail unpurged in the
1450-402 24-well Sample Plate:
3
H: typically 61 %
14
C: typically 95 %
3.4.13.3 Crosstalk
Optical crosstalk due to light piping in the plastic of the sample plates. The values are given for
sample plates with black printing inhibiting crosstalk between the sample wells.
Unquenched sample with a volume of 150 microlitres high flash point cocktail unpurged in the
1450-401 96-well Sample Plate:
In wide 3H window: < 0.05 %
In wide 14C window: typically 1.8 %
Unquenched sample with a volume of 750 microlitres in the 1450-402 24-well Sample Plate:

269

3.4 Specifications

In wide 3H window < 0.02 %

In wide 14C window: typically 0.2 %
3.4.13.4 Stability
Count variation less than 0.5 % / 24 hours (not including random statistics).

3.4.14 Program Specifications

3.4.14.1 Counting protocols
100 fully programmable counting protocols. Start of counting manually through keyboard
command or automatically by the use of ID labels.
Delay Start, allows for sample incubation prior to start of measurement.
Edit, Copy, Print, Delete, List features for management of protocols.
Password protection against unwanted changes of protocol.
Help function, built-in operating instructions supports the user throughout the program.
Single label CPM or DPM counting.
Dual label CPM or DPM counting.
Crosstalk correction to allow different types of microtitration plate to be used.
Scintillation proximity assay can be done.
Preset parameter for 6 common isotopes.
Count termination by counting time or reached precision (2 sigma error).
Plate counting control, whole plate or only active rows or positions.
Default, Short or Long printer output for quick printout settings.
Fully programmable printout selection. Including statistical analysis of results and user
functions.
Fully programmable output to display. Including statistical analysis of results and user
functions.
270

3.4 Specifications

Fully programmable output to mainframe. Including statistical analysis of results and user
functions.
Fully programmable output to file on terminal PC. Including statistical analysis of results and
user functions.
Repeat or Replicates counting.
Cycle counting, repeat counting of the whole assay.
Background correction. CPM/DPM results can be corrected for experimental background
either by typed in background value or background samples.
Programmable counting time for background samples.
Half-life correction, corrects the CPM/DPM values for decay of short-lived isotopes. Zero time
start of assay or specified date and time.
Chemiluminescence correction, corrects the CPM/DPM values for random coincidence
contributions.
Statistics Monitor, detects distorting contributions from static electricity.
Static electricity discharge is a phenomenon which occurs together with the use of plastic
sample carriers such as microtitration plates.
3.4.14.2 Normalization protocols
Up to 100 Normalization protocols/data sets. Normalization of the detectors is required to
eliminate possible difference in counting efficiency between the detectors. Normalization is
done for the different isotopes, different applications or different window settings. The
Normalization protocol to be used is selected in the counting protocol.
Also available in the protocol are:
Half-life correction.
Chemiluminescence correction.
Password protection.

271

3.4 Specifications

3.4.14.3 Standardization protocols

Up to 100 Quench standardization protocols/data sets. Quench standardization is needed to
correct for counting efficiency variation or quench in the samples. The Quench Standardization
protocol to be used is selected in the counting protocol.
Selection of curve fit method; smoothed spline, interpolation spline, linear interpolation or
linear regression.
Input of individually adjusted DPM values for the standard samples.
Also available in the protocol are:
Chemiluminescence correction.
Half-life correction.
Password protection.
3.4.14.4 MicroBeta JET injector system
One injector module dispensing at a time. Reading before dispensing, reading after dispensing
and delay time after dispensing.

3.4.15 Consumables
3.4.15.1 Sample Plates
1450-401 96-well Sample Plate (25/box): 96 round-bottomed wells, 8x12 format, made of
clear PET-plastic, printed lines between wells, chemically resistant to all HiSafe-cocktails,
max. volume 250 L/well. Sealed with tape type 1450-461 or 1450-462 or heatsealed with
1450-463 (in 1495-021 Microsealer). Counted in cassette type 1450-101.
1450-402 24-well Sample Plate (25/box): 24 flat-bottomed wells, 4x6 format, made of clear
PET-plastic, printed lines between wells, chemically resistant to all HiSafe-cocktails, max.
volume 1 mL/well. Sealed with tape type 1450-461 or 1450-462. Counted in cassette type
1450-102.
1450-405 96-well Sterile Sample Plate (10/box): 96 flat-bottomed wells, 8x12 format, made
of polystyrene, white well walls and clear well bottom, sterile tissue culture-grade, with a lid,
max. volume 350 L/well. For tissue culture samples and for samples with Polystyrene Safe
cocktail 1450-444. Sealed with tape type 1450-461 or 1450-462. Counted in cassette type
1450-105.

272

3.4 Specifications

1450-407 96-well Sample Plate, PS, 400 L (25/box): 96 flat-bottomed wells, 8x12 format,
made of clear polystyrene, max. volume 400 L /well. For aqueous samples (SPA) or for
samples with Polystyrene Safe cocktail 1450-444. Sealed with tape type 1450-461 or 1450462. Counted in cassette type 1450-103.
1450-408 24-well Heat-sealable Sample Plate (25/box): 24 flat-bottomed wells, 4x6 format,
made of clear PET-plastic, printed lines between wells, chemically resistant to all HiSafecocktails, max. volume 1 ml/well. Sealed with tape type 1450-461 or 1450-462 or heatsealed
with 1450-463 (in 1495-021 Microsealer using 1495-023 Heat Seal Support 24). Counted in a
cassette 1450-102.
1450-410 96-well Rigid Sample Plate (25/box): 96 flat-bottomed wells, 8x12 format, made of
polystyrene, white well walls and clear well bottom, max. volume 350 L/well. For aqueous
samples (e.g. SPA) or for samples with Polystyrene Safe cocktail 1450-444. Sealed with tape
type 1450-461 or 1450-462. Counted in cassette type 1450-105.
1450-419 ScintiStrip (120/box): 12 breakable strips of 8 wells on a support frame 1450-482,
made of scintillation plastics, max. volume 300 L/well. For immobilised samples or aqueous
samples, no scintillation cocktail needed! Sealed with tape type 1450-461 or 1450-462.
Counted on the support frame in cassette type 1450-105.
1450-514 96-well Isoplate (25/box): 96 flat-bottomed wells, 8x12 format, made of clear
polystyrene wells with a rigid white frame, max. volume 350 L /well. For aqueous samples
(e.g. SPA) or for samples with SuperMix cocktail 1200-439. Sealed with tape type 1450-461 or
1450-462. Counted in cassette type 1450-105.
3.4.15.2 Filtermats
1450-421 Printed Filtermat A (100/box): 96-position glassfibre filter mat with printed
pattern. For harvested cell and receptor samples. Scintillant to be used: Betaplate Scint 1205440 or MeltiLex B/HS 1450-442. Placed in a sample bag type 1450-432. Counted in cassette
type 1450-104.
1450-422 Filtermat A (100/box): 24-position glass -fibre filter mat with printed pattern. For
harvested cell and receptor samples. Scintillant to be used: Betaplate Scint 1205-440 or
MeltiLex A 1450-441. Place in a sample bag 1450-432. Counted in cassette type 1450-116.
1450-423 Printed Nylon Membrane (50/box): 96-position nylon membrane with printed
pattern; for DNA samples. Scintillant to be used: Betaplate Scint 1205-440 or MeltiLex A
1450-441. Placed in sample bag type 1450-432. Counted in cassette type 1450-104.

273

3.4 Specifications

1450-424 Filtermat B (50/box): 24-position filtermat with printed pattern made of glassfibre.
For harvested or pipetted samples. Scintillant to be used: Betaplate Scint 1205-440 or MeltiLex
B/HS 1450-442. Place in a sample bag 1450-432. Counted in cassette type 1450-116.
1450-521 Printed Filtermat B (50/box): 96-position glassfibre filter mat with printed pattern.
For harvested receptor or cell samples. Scintillant to be used: Betaplate Scint 1205-440 or
MeltiLex A 1450-441. Placed in sample bag type 1450-432. Counted in cassette type 1450104.
1450-522 DEAE Filtermat (100/box): 96-position filter mat with printed pattern made of
glassfibre, containing positively charged groups. For harvested or pipetted (negatively charged)
samples. Scintillant to be used: Betaplate Scint 1205-440 or MeltiLex A 1450-441. Placed in
sample bag type 1450-432. Counted in cassette type 1450-104.
1450-523 P30 Filtermat (100/box): 96-position filter mat with printed pattern made of
glassfibre, containing negatively charged groups. For harvested or pipetted (positively charged)
samples. Scintillant to be used: Betaplate Scint 1205-440 or MeltiLex A 1450-441. Placed in a
sample bag 1450-432. Counted in a cassette type 1450-104.
3.4.15.3 Plastic Bags
1450-431 Waste Bag (100/box): Waste bag for used sample plates. Chemically resistant to
safe cocktails. Closed with a heatsealer (1295-012).
1450-432 Sample Bag (100/box): Sample bag for filters 1450-421 and 1450-423. Chemically
resistant to safe cocktails. Closed with a heatsealer (1295-012).
3.4.15.4 Scintillation Products
1450-419 ScintiStrip (120/box): 12 breakable strips of 8 wells on support frame type 1450482, made of scintillation plastics, max. volume 300 L/well. For immobilised samples or
aqueous samples, no scintillation cocktail needed! Sealed with tape type 1450-461 or 1450462. Counted on the support frame in cassette type 1450-105.
1450-441 MeltiLex A (100/box): Meltable solid scintillator for filters 1450-421 and 1450-423
or other filters. Melting temperature +60..+120oC.
1450-442 MeltiLex B/HS (50/box): Meltable solid scintillator for filters 1450-521, 1450-424
and other thick filters. Melting temperature +60..+120oC.
1450-444 Polystyrene Safe (5L/box): Scintillation cocktail to be used with polystyrene sample
plates, e.g. 1450-405, 1450-407 and 1450-410.

274

3.4 Specifications

3.4.15.5 ID Products
1450-451 ID Support Plate: Support plate for ID labels. Placed onto a counting cassette.
1450-452 ID Labels (10 sheets/box): Labels for identifying Assay number (necessary for
MultiCalc assays), Protocol and Cassette number and Function code. Fixed on a Support Plate
1450-451.
3.4.15.6 Tapes
1450-461 Sealing Tape (100/box): Sealing tape for sample plates, permanent.
1450-462 Removable Sealing Tape (100/box): Sealing tape for sample plates; removable.
1450-463 Heatsealing Foil (100/box): Sealing foil for sample plates type 1450-401 and 1450408. Heatsealed with a 1495-021 Microsealer.
1450-465 Printed Sealing Tape (100/box): Sealing tape for sample plates, printed.
1450-466 Coloured Sealing Tape (100/box): Sealing tape for luminescent sample plates.
3.4.15.7 Other Consumables
1450-481 Support Frame (25/box): Support frame for sample plates type 1450-401, 1450-402
and 1450-408. Removed before counting.
1450-482 Support Frame for 1450-407 (25/box): Support frame for sample plates type 1450407 and 1450-419. Removed before counting 1450-407. 1450-419 ScintiStrips are counted
with the support frame.
1450-486 MicroBeta Starter kit, MicroPlates: Contains the MicroBeta sample plates (5 of
each), tapes and heat sealable foil.
1450-487 MicroBeta Starter kit, Filters, Membranes and Solid Scintillators: Contains the
MicroBeta filters, membranes, MeltiLex-sheets, Sample bags and Waste bags.
3.4.15.8 Adapters and Inserts
1450-108 Adapter: For Eppendorf tubes (or equivalent). The adapters are placed on a 1450102 cassette, a bottom tape is used if needed.
1450-109 Insert: For 24-well cell culture plates. The inserts are placed in the sample wells in
order to reduce crosstalk between the wells.

275

3.4 Specifications

276

3.5 Abbreviations and acronyms used

3.5 Abbreviations and acronyms used

.
Sample passed monitor tests (monitor output)
A/D
Analogue to digital
ABS
Absolute value 1
ALT
Alternate key
AQP(I)
Asymmetric quench parameter of the isotope spectrum
AQP(I)% Theoretical percentage error of AQP(I)
AS
Value of assay ID for the current plate 1
ASCII
American Standard Code for Information Interchange
BDH
Laboratory reagent and equipment sales company
BG
Background
BGND
Background
BGNDm Background in window m 2
BIOS
Basic Input Output System
Bk
Background
bps
Bits per second
BS
Backspace key
CA
Value of cassette ID for the current plate 1
CASS
Cassette field in ID support plate
CCPM
Corrected count rate (counts per minute)
CCPM% Theoretical percentage error for CCPM
CCPM1 Corrected count rate (counts per minute) for isotope 1 1
CCPM1% Theoretical percentage error for CCPM1 1
CCPM2 Corrected count rate (counts per minute) for isotope 2 1
CCPM2% Theoretical percentage error for CCPM2 1
CCPM3 Corrected count rate (counts per minute) for window 3 1
CCPM3% Theoretical percentage error for CCPM3 1
CCPS1
Corrected count rate (counts per second) for isotope 1 1
CCPS2
Corrected count rate (counts per second) for isotope 2 1
CCPS3
Corrected count rate (counts per second) for window 3 1
CGA
Colour Graphics Adapter
ch
Channel
CHISQ1 Reduced chi-square value for CCPM1, DPM1 or LCPS 1
CHISQ2 Reduced chi-square value for CCPM2 or DPM2 1
CLM%
Chemiluminescence in percentages in window 1 1
CNO
Cassette order number 1
COUNTS1 Counts for isotope 1 1
COUNTS2 Counts for isotope 2 1
COUNTS3 Counts for window 3 1

277

3.5 Abbreviations and acronyms used

cpi
Characters per inch
CPM
Count rate (counts per minute)
CPM1
Count rate (counts per minute) for isotope 1 1
CPM1% Theoretical percentage error for CPM1 1
CPM2
Count rate (counts per minute) for isotope 2 1
CPM2% Theoretical percentage error for CPM2 1
CPM3
Count rate (counts per minute) for window 3 1
CPM3% Theoretical percentage error for CPM3 1
CPM_C Coincidence count rate (counts per minute)
CPM_NC Non-coincidence count rate (counts per minute)
CPM_AQP AQP(I) count rate (counts per minute)
CPS1
Count rate (counts per second) for isotope 1 1
CPS2
Count rate (counts per second) for isotope 2 1
CPS3
Count rate (counts per second) for window 3 1
CT
Cassette type 1
CTIME
Dead time corrected counting time in seconds 1
CTRL
Control key
CV1%
Percentage coefficient of variation for CCPM1, DPM1 or LCPS 1
CV2%
Percentage coefficient of variation for CCPM2 or DPM2 1
D
Detector number 1
DATE
Current date 1
DCCPM1 Absolute theoretical error for CCPM1 1
DCCPM2 Absolute theoretical error for CCPM2 1
DCCPM3 Absolute theoretical error for CCPM3 1
DDPM1 Absolute theoretical error for DPM1 1
DDPM2 Absolute theoretical error for DPM2 1
DEAE
Diethylaminoethyl
DEL
Delete key
DET
Detector number 2
DIN
Di-isopropylnaphthalene
DLCPS
Absolute theoretical error for LCPS 1
DNA
Deoxyribonucleic acid
DPM
Activity (disintegration per minute)
DPM%
Theoretical percentage error for DPM
DPM1
Activity (disintegration per minute) for isotope 1 1
DPM1% Theoretical percentage error for DPM1 1
DPM2
Activity (disintegration per minute) for isotope 2 1
DPM2% Theoretical percentage error for DPM2 1
DPMM
DPM out of range monitor 1
DTR
Flow control using Data Terminal Ready line

278

3.5 Abbreviations and acronyms used

EEC
EFF
EFF1
EFF2
EFFmn
EGA
ERR
ETIME1
ETIME2
EXCL
EXP
FF
FLAG
FU
FUNC
h
HIGH
IAEA
IBM
ID
INT
ISEP
ISEP%
ISO
ISOT
kB
kBq
keV
LAN
LCPS
LCPS%
LM
LN
LSC
m
MAX
MB
MC
MCA
MCGA

European Economic Community

Counting efficiency 2
Counting efficiency for isotope 1 1
Counting efficiency for isotope 2 1
Counting efficiency, window m, isotope n 2
Enhanced Graphics Adapter
Variation after two recounts (statistics monitor output)
Elapsed time in hours for isotope 1 from zero time 1 1
Elapsed time in hours for isotope 2 from zero time 2 1
Controls excluding of unrequired parts of the output 1
Exponent function 1
Formfeed character to the printer after 'End of assay'-message 1
Termination flag 1
Value of function ID code for the current plate 1
Function field in ID support plate
Hour
Too high count rate (luminescence monitor output)
International Atomic Energy Agency
International Business Machines
Identifier
Counting interrupted (termination flag)
Isotope spectrum end point 1
Theoretical percentage error for ISEP 1
International Standards Organization
Isotope
kilobyte
kiloBequerel
kiloelectronVolt
Local area network
Luminescence corrected count rate (counts per second) divided by 100 1
Theoretical percentage error for LCPS 1
LCPS out of range monitor 1
Natural logarithm 1
Liquid scintillation counter
Minute
Maximum counts per channel reached (termination flag)
MicroBeta (starts program from DOS); Megabyte
MultiCalc
Multichannel analyzer
Multicolour Graphics Array

279

3.5 Abbreviations and acronyms used

MEAN1 Mean value for CCPM1, DPM1 or LCPS 1

MEAN2 Mean value for CCPM2 or DPM2 1
MS-DOS Microsoft Disk Operating System
N
No
nCi
nanoCurie
NEWLINE Start a new line 1
NL
Start a new line 1
NO
Number
NOLIVE No status (live) display 1
NORM
Normalization ID label
OUT
Out of range (DPM monitor output)
P30
Ortho phosphate
P1
Probability in percentages for CCPM1, DPM1 or LCPS 1
P2
Probability in percentages for CCPM2 or DPM2 1
PBS
Phosphate buffered saline
PC
Personal computer
PET
Polyethylene terephtalate
PLATE
Output results in microtitration plate format 1
PLATEID Plate identifier
PMT
Photomultiplier tube
POS
Sample position on microtitration plate 1
PR
Protocol number used 1
PREC
Precision reached (termination flag)
PRINTER Output same data as specified for printer 1
PROT
Protocol listing 1; Protocol field in ID support plate
PS/2
Personal System/2
PVC
Polyvinyl chloride
RAM
Random access memory
REC1
Recounted once (statistics monitor output)
REC2
Recounted twice (statistics monitor output)
REPL
Replicate sample range 1
RESP1
Response (CCPM1 or LCPS) 1
RESP1% Theoretical percentage error for RESP1 1
RESP2
Response (CCPM2) 1
RESP2% Theoretical percentage error for RESP2 1
ROM
Read only memory
RP
Repeat number of the current measurement 1
RS-232C Recommended standard number 232, revision C, for data communications
s
Second
S
Spectrum half 1

280

3.5 Abbreviations and acronyms used

S1
SD1
SD2
SEM1
SEM1%
SEM2
SEM2%
SEQ
SH
SPA
SPECTRA
SPSAVE
SQP(I)
SQP(I)%
SQP(I)m
SQRT
STAT
STD
STIME
STM
STOP
TAB
TCA
TCV1%
TCV2%
TIME
TSD1
TSD2
TSEM1
TSEM1%
TSEM2
TSEM2%
TXT
UN
UTMAC
VGA
VT100
VT52
W1
W2
W3

Selects 1 sigma level for all deviation output fields 1

Standard deviation for CCPM1, DPM1 or LCPS 1
Standard deviation for CCPM2 or DPM2 1
Standard error of mean for CCPM1, DPM1 or LCPS 1
Percentage standard error of mean for CCPM1, DPM1 or LCPS 1
Standard error of mean for CCPM2 or DPM2 1
Percentage standard error of mean for CCPM2 or DPM2 1
Running sequential sample number 1
Shelf number 1
Scintillation proximity assay
Plot spectra 1
Save spectra to file in compressed numerical format 1
Spectrum quench parameter of isotope spectrum 1
Theoretical percentage error for SQP(I) 1
SQP(I) for isotope m 2
Square root 1
Data specified will be output when a statistics mean is available 1
Standardization ID label
Counting start time 1
Statistics monitor 1
Stop ID label
Produces tab-character between all output columns 1
Trichloroacetic acid
Percentage theoretical coefficient of variation for CCPM1, DPM1 or LCPS 1
Percentage theoretical coefficient of variation for CCPM2 or DPM2 1
Current time 1
Theoretical standard deviation for CCPM1, DPM1 or LCPS 1
Theoretical standard deviation for CCPM2 or DPM2 1
Theoretical standard error of mean for CCPM1, DPM1 or LCPS 1
Percentage theoretical standard error of mean for CCPM1, DPM1 or LCPS 1
Theoretical standard error of mean for CCPM2 or DPM2 1
Percentage theoretical standard error of mean for CCPM2 or DPM2 1
Produces 'Start of assay:nn'-message 1
United Nations
UltroTerm for Macintosh
Video Graphics Array
Digital Equipment Corporation video terminal type
Digital Equipment Corporation video terminal type
Window 1 limits 1
Window 2 limits 1
Window 3 limits 1
281

3.5 Abbreviations and acronyms used

WT
Wallac terminal emulation
XON/XOFF Flow control using XON (ASCII 17) and XOFF (ASCII 19) characters
Y
Yes
________________________________
1
Programmable output
2
Normalization and standardization tables

282

1390 3693

DECLARATION OF CONFORMITY FOR CE-MARKING

We
Supplier's name

WALLAC OY
Address

PL 10, 20101 TURKU, FINLAND

declare under our sole responsibility that the product
Name, type or model, lot, batch or serial number, possibly sources and numbers of items

1450 MicroBeta TriLux

1450 MicroBeta JET

Valid from serial number 4500762

Valid from serial number 4500877

to which this declaration relates is in conformity with the following standard(s) or other normative
document(s)
Title and/or number and date of issue of the standard(s) or other normative document(s)

EN 50082-1 :1992; EN 50081-1 :1992

EN 61000-3-2 : 1995 + A1:1998 + A2:1998 + A14:2000; EN 61000-3-3:1995
EN 61010-1 :1993

(if applicable) following the provisions of the following directives

Electromagnetic compatibility (EMC), 89/336/EEC

Low voltage (LV), 73/23/EEC

Date and place of issue

27 December 2000 TURKU, FINLAND

Name and signature or equivalent marking of authorized person

Pekka Mkinen, Quality Assurance Manager

283

284

Part 4 Installation information

285

286

4.1 Installation instructions

4.1 Installation instructions

Note: Except for where noted these instructions are valid for both MicroBeta TriLux and
MicroBeta JET.

4.1.1 Environment
Although normal clean laboratory conditions are usually quite satisfactory as an operational
environment it is useful to take the following points into consideration.
If possible a separate room should be provided for the MicroBeta LSC as this allows the best
control over the immediate environment. Ventilation in the room should be adequate for all
conditions of use, the temperature should be reasonably constant at about 22oC, relative
humidity should not be excessive, and direct sunlight should not be able to reach the
instrument. It is also important that the various isotopes are stored well away from the
instrument in another room. Only those radioactive samples that are actually measured should
be in the laboratory at any time in order to keep the background at a low level.

4.1.2 Electric power

Three electrical outlets each having a protective earth should be available, with, if possible, a
separate power line for the instrument itself having an isolation switch and a fuse box. If
excessive fluctuations in the mains voltage are anticipated, a mains stabilizer may be
necessary.

287

4.1 Installation instructions

4.1.3 Unpacking

Removing
MicroBeta from
its packaging

Cut off and remove the binding bands

Open the clinching nails and lift the cover off from the base of the package, see the figure in
the previous column.
Unpack all units and accessories and check them according to the packing list also noting any
possible transport damage.
Open the lower clinching nails and remove the packing around the main instrument.
Lift the instrument up from the bottom of the package.
Move the instrument to its place of operation.

288

4.1 Installation instructions

4.1.4 Checking the mains voltage setting

Measure and note the mains voltage at the outlets to be used.
Locate the mains selector switch, this is on the right side when looking from the
rear of the instrument. If necessary adjust the mains selector switch to correspond
with the measured supply. For supplies with a nominal voltage of 230 V it is
recommended that the selector be set to 240 V.
Check that the fuses fitted in the fuse carriers on the back panel are of the correct rating for the
local supply, and according to the label - T 4 A for supplies with a voltage in the range 100 to
120 V and T 2 A for supplies with a voltage in the range 220 to 240 V. For continued
protection against fire hazard, replace only with the same type and rating of fuse.
Insert the battery fuse in its holder into the socket at the rear of the instrument.

4.1.5 Connecting up the counter and peripherals

Connect the counter to the PC and the printer using the cables shown in the figure.

289

4.1 Installation instructions

There are two configurations, either the printer and PC are each connected to MicroBeta to
ports 3 and 1 respectively (not possible with MicroBeta JET) or the printer is connected to the
PC which is in turn connected to port 1. This latter configuration is for use with the Windows
workstation and MultiCalc.
See the additional section at the end of the chapter for information about Installation of the
Injector system to MicroBeta JET.
Plug in the power cables for each device.

4.1.6 Start-up
Although the UltroTerm terminal emulator program is the default emulator for MicroBeta,
other emulators, e.g. GenTerm and MultiCalc, or VT52 and VT100 terminals can also be used.
The following two sections describe how to start MicroBeta with the default UltroTerm and
with another emulator.
4.1.6.1 Start-up using UltroTerm
This is the default setting and therefore no parameters must usually be changed in the counter.
1. Switch on the printer.
2. Switch on the PC.
3. Install UltroTerm as described in the UltroTerm User Manual and in section 2.31.5.3
Installation in this manual.
4. Start UltroTerm as described in the UltroTerm User Manual and in section 2.31.5.4 Starting
UltroTerm in this manual.
5. If necessary, change the communication parameters as described in the UltroTerm User
Manual and in section 2.31.5.5 in this manual.
6. Insert the MicroBeta program disk into drive A:, the lower disk drive of the counter.
7. Insert the MicroBeta protocol disk into drive B:, the upper disk drive of the counter.
8. Switch on the counter with the power switch on the back of it.
9. After about 3 minutes Ready is displayed and the counter is ready for operation.
290

4.1 Installation instructions

4.1.6.2 Start-up using another terminal emulator

When the program is started it will check what kind of terminal or emulator is connected to the
counter and change its own settings according to this. It will also test the baud rate and the
number of data bits and make necessary changes. The program can identify the same emulators
as are available in the counter, i.e. GenTerm WT, UltroTerm, MultiCalc VT52, standard VT52
and VT100. It can also identify baud rates from 300 to 9600 bps and 7 or 8 data bits.
Together with changing emulator and communication parameters, the program will also notify
the user that the change has been made and that the user must check and correct the terminal
and/or RS settings in System parameters. The program will usually work fine with the
automatic settings, but it is recommended that the user checks and corrects the system
parameters.
1. Switch on the printer.
2. Switch on the PC.
3. Start your terminal emulator.
4. If necessary, change the communication parameters in the terminal emulator.
5. Insert the MicroBeta program disk into drive A:, the lower disk drive of the counter.
6. Insert the MicroBeta protocol disk into drive B:, the upper disk drive of the counter.
7. Switch on the counter with the power switch on the back of the counter.
8. After about 2 minutes a message like the following will appear: Incorrect terminal
type. GenTerm (or MultiCalc, VT52 or VT100) used. Please select terminal
type in (S)ystem/Ter(m)inal.
9. Press the S key and the System menu is displayed.
10. Press the M key to change terminal parameters.
11. Change to your terminal emulation by entering the number followed by Enter.
12. If necessary, change the terminal parameters as described in section 2J.30.10
System/Terminal in this manual.
13. Press any key, e.g. the space bar, when you are told to change emulator.

291

4.1 Installation instructions

14. The program will return to the System state.

15. If necessary, change the RS communication parameters as described in section 2J.30.7
System/RS-232C in this manual.
16. Press the Q key.
17. Ready is displayed and the counter is ready for operation.
4.1.6.3 Troubleshooting
If there seems to be communication problems, e.g. characters are displayed but the keyboard
does not function, it is probably one of following errors:
Wrong terminal emulator
Example: The counter is configured for GenTerm WT emulation but the PC uses UltroTerm.
Solution: When starting the program, the counter will usually check the emulator in the PC and
change its own settings according to this. If this does not work then proceed as follows:
Restart the counter by putting the power switch off and on again. When in the Ready state,
press the Esc key (F3 in UltroTerm) one or several times. The counter should respond Please
check terminal type in (S)ystem/Ter(m)inal!. After this the keyboard will work and
you can change the terminal emulation in System/Terminal (press S followed by M). See
chapter 2.31 Terminal emulators for further explanations.
Error in communication parameters
Example: The RS-232C communication parameters in the counter differ from those in the PC.
Solution: Set the communication parameters to be the same in the counter and PC. If you are
changing the parameters in the counter then restart it as shown above. Go to System/RS-232C
(press S followed by R) and set the parameters as described in section 2J.30 System/RS-232C.
See section 2.31 Terminal emulators/GenTerm/Changing parameters in GenTerm or 2.31
Terminal emulators/UltroTerm/Changing parameters in UltroTerm for how to change the
parameters in GenTerm or UltroTerm emulators, or see the respective manual.
4.1.6.4 Detector usage/shelf usage
In case of a 1, 2 or 3 detector model of MicroBeta, the detector usage must be defined in the
system parameters before starting counting. Go to the (S)ystem state and select the (l)evel
(T)esting. Then select Detector/shelf (u)sage and choose one of the five detector alternatives.

292

4.1 Installation instructions

Check also that the shelf usage (1, 16 or 32 shelves) mask type, ParaLux counting and Plate ID
reader are set correctly. Here you can also set the selected detector settings to all protocols. Use
this feature with care.

4.1.7 Functional check

4.1.7.1 Preparing a functional check
Carry out a functional check of the instrument, including
checking the function of the ID reader. To do this
proceed as follows:
Prepare four cassettes with labels. On the first cassette
use labels 1 - 3 and NORM in the function field (Assay =
1, Protocol = 2, Cassette = 3 and Function code =
NORM), on the second 4 - 6 and STD, on the third 7 - 9
and STOP and on the fourth 10 - 12 with the function
field empty. See chapter 2.4 for details of coding of a
cassette. Insert the first cassette into shelf 1, (see the
figure on the right), the last into shelf 16 (32) and the
other two somewhere between.
4.1.7.2 Starting the functional check
The ID test is started in the Conveyor state on the test
level. To get this go to System by pressing S, select
Level by pressing L (this does not appear on the
computer display but the figure on the next page shows
it). Then select the Test level by pressing T. Go back to
the Ready state by pressing Q and there select V (see the
example below). Then press T and enter the number of
runs. The instrument will run the cassettes once and print
the results. After this it only prints ID read errors. Keep
the door open and check that the instrument is running
alright mechanically.
NOTE! Keep your fingers away from the cassette rack
when the door is open!

293

Last
cassette

First
cassette

4.1 Installation instructions

Ready>V
(H)elp
(C)lear conveyor
(N)ext cass
(G)oto pos
Next le(v)el
Calib (0)-pos
(F)ocus ID reader
(.)Park det
Di(s)pOut

(Q)uit
Rack (u)p
(P)rev cass
Si(z)e checkpoint
Reset r(a)ck
Calib det bloc(k)
Print ever(y) ID
Shelf (1)..(16)

(O)ff
Rack (d)own
Cu(r)rent cass
C(l)ose detector
(I)nit mask
(J)ust read IDs
Print ra(w) IDs
(*)Calib sensor

Remove ca(s)s
Ne(x)t pos
Op(e)n detector
Change (m)ask
(T)est IDs
Print (b)ad IDs
Di(s)pIn

Conveyor shelf 1>T

Number or runs 100 ->5
Assay: 1
Prot:
Assay: 4
Prot:
Assay: 7
Prot:
Assay: 10
Prot:
Conveyor stopped
ID test results:

2
5
8
11

Cass:
Cass:
Cass:
Cass:

3
6
9
12

Func:
Func:
Func:
Func:

NORM
STD
STOP
NO ID

Shelf:
Shelf:
Shelf:
Shelf:

1
4
11
16

Runs: 5
Errors: 0
Read once: 20, twice: 0, thrice: 0

The text above should be printed if the test is OK. Check that there are not many occasions
when the ID label is read twice or thrice because this shows that the ID reader is performing
poorly.
4.1.7.3 Test settings
For checking the performance of the instrument, run the detector efficiency normalization with
the normalization standard plate supplied with the instrument. Attach the printout of the
measurement to the installation report.
4.1.7.4 System operation showing the TEST level
The figure on the next page is a flow diagram of the whole MicroBeta operating system. It
shows how the main control letters (in parentheses) lead to the various functions and how the
test functions are related to the normal functions. Compare with the flow diagram in 2J.1.5
showing the functions on the normal level.
4.1.7.5 Leaving the TEST level
To return to the normal level press Q to return to the Ready state, then S to go to the System
state. Press L for level selection and N for normal level (not shown). Press Q to return to the
Ready state.

294

4.1 Installation instructions

Flow chart of MicroBeta commands (service level)

Info >
(H)elp
(Q)uit
(U)sage
(C)onsumables
(P)ositions
Ch(a)nges
Cu(s)tomizing
S(y)stem

Ready >
(H)elp
(I)nfo
(C)ount
(P)rotocols
(S)ystem

System >
(H)elp
(Q)uit
MS-D(O)S
(E)rror beep
(S)tatus display
(V)ersion
(C)lock
Serial (n)o
Temp cali(b)
(*)HV On/Off
Detector (u)sage
Se(t)up mode
(R)S-232C
(P)rinter
(D)ata drive
Ter(m)inal
P(a)sswords
Customi(z)ing
(-)Detector temp.

Total count rate>

(H)elp
(Q)uit
(S)how
(P)rint
(R)eset
(D)efine efficiencies
Injector> (JET only)
(H)elp
(Q)uit
(I)nit
(D)ispense
(P)rime
(T)ip
(W)ash
Suck-(b)ack
Co(m)mand
(S)tatus
O(f)f
(N)ext pos
(G)oto pos
(C)lear conveyor
Disp(e)nser IN/OUT

Count >
(H)elp
(Q)uit
(nn) (count.prot.no)
(?)
(Nnn) (norm.prot.no)
(N?)
(Dnn) (std.prot.no)
(D?)
(A)utomatic counting
Operate con(v)eyor
(T)otal count rate
(M)annual counting
Injector (X)L

Protocols >
(H)elp
(Q)uit
(C)ounting protocol
CPM (n)ormalization prot.
DPM (s)tandardization prot.
(M)ultiCalc assay protocol

MicroBeta A:\ >

MB - Back to Ready >
MultiCalc assay protocol >
(H)elp
(Q)uit
(S)how
(P)rint
(L)ist

DPM stand. protocol>

(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
(R)eplot

295

Counting protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
CPM norm. protocol>
(H)elp
(Q)uit
(E)dit
(nn) (= prot.no)
(D)elete
(S)how
(P)rint
(L)ist
De(f)ault
(C)opy
(N)ormalization data

Conveyor>
(H)elp
(Q)uit
(O)ff
(C)lear conveyor
Rack (u)p
Rack (d)own
Remove ca(s)s
(N)ext cass
(P)rev cass
Cu(r)ent cass
Ne(x)t pos
(G)oto pos
C(l)ose detector
Op(e)n detector
Next le(v)el
Reset r(a)ck
(I)nit mask
Change (m)ask
(T)est IDs
(F)ocus ID reader
(J)ust read IDs
Print ever(y) ID
Print ra(w) IDs
Print (b)ad IDs
Si(z)e checkpoint
Calib (0) -pos
Calib det bloc(k)
(.)Park det
Shelf(1)..(16)/(32)
D(i)spIn#
Di(s)pOut#
(*)Calib sensor

Sensors>
(H)elp
(Q)uit
(Y) Rack
(X) Rack
XY(Z)ero
XYPos(F)ine
(D)etUp
Det(U)pPos
(P)arkDet
(E)dit

4.1 Installation instructions

4.1.8 Setting the system parameters

During the installation some system parameters should be checked and set if required. These
parameters can be set in the System-state of the instrument program (see chapter 2J.30
System). A checklist of the system parameters is provided in what follows:
4.1.8.1 Terminal emulation
Instrument Manual:
- See chapter 2J.30 System/Ter(m)inal.
Default:
- UltroTerm terminal emulator program.
- 23 lines on screen
Depending on which terminal emulation the user wants to use, check the following:
4.1.8.1.1 GenTerm in WT emulation.
GenTerm is a terminal emulator program supported by MicroBeta. The following parameters
must be set if GenTerm is to be used:
(a) Set Terminal emulation to 1=GenTerm.
(b) Set Number of lines on screen at maximum to 24.
Notes:
1: GenTerm V2.B or later is required.
2: See chapter 2.31 Terminal emulators/GenTerm/Installation.
3: See also 4.8.2 and 4.8.5.
4.1.8.1.2 UltroTerm
This is the default terminal emulation and so normally you need not change the terminal
emulation settings. However if the terminal PC is some portable model that is not capable of
displaying 23 lines then change the Number of lines on screen to the maximum the PC can
display.

296

4.1 Installation instructions

Notes:
1: UltroTerm V2.00 or later is required.
2: See the sections 2.31 Terminal emulators/UltroTerm/Installation and Terminal
emulators/GenTerm/ Troubleshooting.
3: See also 4.8.2 and 4.8.5.
4. 1.8.1.3 MultiCalc
(a) Set Terminal emulation to 3= MultiCalc.
(b) The MultiCalc installation procedure sets other terminal features.
Notes:
1: MultiCalc V1.00 or later is required together with a 1450 communication protocol.
2: See chapter 2.22 MultiCalc and chapter 2.31 Terminal emulators/ GenTerm /Trouble
shooting.
3: See also 4.8.2 and 4.8.5.
4.1.8.1.4 Other terminal emulator programs or terminals
MicroBeta can also support other terminal emulator programs and pure terminals that use
VT52 or VT100 terminal emulation. If the user wants to utilize one of these then see section
2J.30 System/Ter(m)inal for the possible settings.
Notes:
1: See chapter 2.31 Terminal emulators/VT52 and VT100 compatible terminals about using
VT52/VT100 type terminals.
2: See also GenTerm/Troubleshooting in the same chapter.
3: See also 4.8.2 and 4.8.5.

297

4.1 Installation instructions

4.1.8.2 Printer connection

Instrument Manual:
- See chapter 2J.30 System/Ter(m)inal.
Default:
- The printer is connected to the terminal PC parallel printer port.
There are two possible ways to connect the printer:
4.1.8.2.1 Printer connected to the instrument RS232C port 3
See the manuals belonging to the printer about how to set up the printer for use with a serial
interface.
Go to the Ter(m)inal selection in System-state and set the line Print through terminal to N=No
(do not change other lines).
Note: See 4.8.6 for the default RS232C protocol and how to change it.
4.1.8.2.2 Printer connected to the terminal PC parallel/printer port
As this is the default connection there is no need to change system parameters.
Proceed as follows:
(a) Use a standard Centronics-interface cable to connect the printer and the terminal PC.
(b) See the manuals supplied with the printer about how to set up the printer for use with a
parallel interface.
4.1.8.3 Printer type
Instrument Manual:
See chapter 2J.30 System/(P)rinter.
Default:
- Epson FX with printing resolution of 12 characters per inch (cpi).

298

4.1 Installation instructions

The printer type selection affects only graphical output (e.g. spectrum plots) and the automatic
printing resolution setting (cpi). The printing resolution affects the possible line lengths of
outputs (10 cpi = 80, 12 cpi=96 and 17 cpi=136 character per line on a standard 8 inch paper).
The printer types supported are:
4.1.8.3.1 IBM graphics printer
Select this only if the printer does not support Epson FX emulation but supports IBM graphics
printer emulation instead. This mode does not support 12 cpi mode.
4.1.8.3.2 Epson FX
As this is the default mode you need not change printer type if the printer supports this
emulation.
4.1.8.4 Clock setting
Instrument Manual: - See 2J.30 System/(C)lock.
Check the date and time and set them to the current values if needed.
4.1.8.5 Data drive
Reference Manual: - See chapter 2J.30 System/(D)ata drive.
Default: - Drive A:.
Select the diskette/hard disk drive and the directory for the MicroBeta result files as described
in the Instrument manual. Depending on the terminal emulation set in section 4.1.8.1. you must
do the following:
4.1.8.5.1 GenTerm
No need for extra settings.
Note: See chapter 2.31 Terminal emulators/GenTerm/Result files.
4.1.8.5.2 UltroTerm
UltroTerm itself takes care of result file saving.
Note: See chapter 2.31 Terminal emulators/UltroTerm/Result files.
4.1.8.5.3 MultiCalc
MultiCalc itself takes care of result file saving.

299

4.1 Installation instructions

Note: See chapter 2.22 MultiCalc and the MultiCalc User Manual chapter on files.
4.1.8.5.4 Other terminal emulator programs or terminals
As commercial terminal emulators differ a lot it is not possible to give any advice which covers
all of them.
Note: See the chapter 2.31 Terminal emulators/VT52 or VT100-compatible terminals/Result
files.
4.1.8.6 RS-232C-interface protocol
Instrument Manual: - See chapter 2J.30 System/(R)S-232C.
Default (DTR means electronic DTR handshake protocol):
- Port 1=Terminal: 9600 baud, 8 data bits, 2 stop bits, no parity, DTR
- Port 2=External: 9600 baud, 8 data bits, 2 stop bits, no parity, DTR
- Port 3=Printer : 4800 baud, 8 data bits, 2 stop bits, no parity, DTR
Normally these need not be changed. However some situations when they must be are:
(a) Port 1=Terminal: If the terminal PC is going to be used for other purposes during counting
(MultiCalc or UltroTerm allow this) then it is recommended to lower the baud rate to 1200
baud or lower to ensure that characters will not be lost.
(b) Port 2=External computer: The interface can depend very much on the external computer
used.

4.1.9 Parking the detectors

If the instrument needs to be moved from the place where it was originally installed it is
recommended that the detectors should be parked during the time when the instrument is
actually being moved. This can be done as follows:
Go to the (S)ystem state and select (L)evel and (T)est. Press Q to return to the Ready state and
then O to operate the conveyor. Press . (period). This will move the upper detector block
against the lower one so that it cannot shift when the instrument is moved. The following
message will be seen on the screen: Detectors are now parked. Please turn the power
off. When the instrument is restarted the detector blocks will separate again automatically.

300

4.2 Installation of the Injector System

4.2 Installation of the Injector System to MicroBeta

JET
Make a normal MicroBeta installation (mechanical check and check of efficiencies and
background). Switch the instrument off and attach the injector system to the instrument:
-attach the injector modules to the base (two screws at the bottom)
-attach the support pieces for the solution bottles
-attach the transparent tubing of the solution bottles to the IN connectors of the pumps
-attach the black injector adapter tube No. 1 to the out connector of pump No. 1 and tube No. 2
to the out connector of pump No. 2 etc. If there is more than one injector module the units are
numbered.

Injector Module

Axle attached to
the instrument

Solution
Bottles
IN
Base
Injector assembly
OUT

301

4.2 Installation of the Injector System

-attach the whole injector system to the left side of the MicroBeta JET by sliding the axles into
holes in the support pieces attached to the instrument.

302

4.2 Installation of the Injector System

- open the lid of the injector adapter module and check the tubing (see fig below). Systems with
two injector modules have a second tube on the left side connector

-cut the tie which is attached to the arm for the injector assembly

303

4.2 Installation of the Injector System

-attach the injector assembly to the arm (see fig below).

304

4.2 Installation of the Injector System

-tie the tubing to the arm motor so the tubing can freely move into the instrument when
injecting (see fig below).

305

4.2 Installation of the Injector System

-connect the cable from the injector system to the connector at the rear of MicroBeta JET (see
fig below)

Boot the system up and check the maintenance section of the User Manual, page 16.
After initializing and priming you should dispense at least once the volume to be used in the
assay. The dispensing volumes and alignment to the sample plate can be tested by running a
counting protocol. Counting protocol parameters are set for the dispensing speed and volumes
for the selected modules.

306

Part 5 Index

307

308

5 Index

5 Index
Program diskettes, 58
Protocol diskettes, 58
Terminal diskette, 58
Barcode, 29
Barcode reader, 250
Baud rate, 148, 234
Block graphics, 153, 239
Blocked transportation system
Loading cassettes, 78

/
/, 16, 188

?
?
Help, 73

@ in path name, 49, 120, 152, 225, 238

Calculation methods, 255

Capacity for samples, 265
Cassette number, 29
Cassette type, 116, 221
Cassettes, 25, 266
Coding, 29
Stop, 30
Change parameters
GenTerm, 162
Change special features, 121, 225
Changes, 74
Changing parameters
UltroTerm, 166
Channel (injector), 251
Chemiluminescence correction, 122, 129, 226, 260
Chi-square test
Mean value, 263
Chromatography, 89
Chromium release, 89
Cleaning, 253
Clear conveyor, 186
Clock, 27, 148, 234
Coding, 250
Coding cassettes, 29
Coding filtermats, 31
Colour quenching and SPA, 89
Command, 186
Communication drivers, 95
Communication loss, 149, 235
Connections, 265
Consumables, 74, 272
Adapters and inserts, 275
Filtermats, 273
Heatsealing foil, 275

A
Abbreviations used, 277
Acronyms used, 277
Active rows, 116, 221
Activity, 259
Activity standardization, 259
Adapters, 275
Adjusted activities (stand.), 126, 230
AQP(I), 61, 63, 115, 219, 259
Aspirating tube I.D., 244
Aspirating tube length, 244
ASQP(I)
Quenching, 63
Assay protocol number, 30
Assay protocols, 97
Asymmetric quench parameter, 61, 259
Automatic operation
Counting (terminal operation), 33
Counting with MultiCalc, 34
Normalization, 33
Standardization, 33
Automatic result file deletion, 154, 240
Autoquench correction, 123, 227

B
Background, 268
Background - thermal, 106, 210
Background correction, 121, 129, 226, 256
Backslash, 16, 188
Backspace, 16, 188
Backup

309

5 Index

ID products, 275
MeltiLex A, 274
Miscellaneous, 275
Plastic bags, 274
Polystyrene Safe, 274
Sample plates, 272
ScintiStrip, 273, 274
Tapes, 275
Control keys, 17, 188
Control-k
Hiding text, 112, 216
Controlling MicroBeta, 15, 187
Costar plate, 123, 227
Count all position in cassette, 116, 221
Count rate, 255
Corrected, 256
Counter, 95
Counting, 33
Control, 37, 121, 197, 225
During, 34
Mode, 114, 218
Next row (N), 35
Next assay (E), 35
Off (O), 35
Starting, 33
Stopping, 35
Time, 28, 37, 116, 197, 220
Time for background (norm), 124, 228
Time for standards (norm), 124, 228
CPM counting, 61
Crosstalk, 269
Crosstalk correction, 41, 123, 125, 228, 229
CPM normalization, 41
DPM standardization, 44
Curve fit method selection (stand.), 125, 230
Customi(z)ing, 155, 241
Customizing, 74
CV
Mean value, 262
Cycles, 38, 198

Processing, 51
Saving, 50
Data bits, 149, 235
Data drive, 150, 236
Datafiles, 49
Accessing, 50
External, 51
In MultiCalc, 51
Names, 50
Saving data, 50
Date, 27
Decay correction
Half-life, 71
DEL key, 16, 188
Delay between plates, 39, 123, 200, 227
Delay time after dispensing, 220
Delayed start, 34
Detector assembly, 266
Detector efficiency
Correction, 257
Normalization, 255
Detector usage (1, 2 or 3)
Start up, 292
Detectors, 53, 250
Number of, 53
Detectors - parking, 300
Di-isopropylnaphthalene, 87
Dimensions, 265
DIN, 87
Discriminator channel, 115, 219
Disk drive
Selection, 49
Diskettes
Disks, 57
Disks
Backup, 58
Counter, 57
Handling, 57
Preparing protocol disk, 59
Program, 57
Program disk, 8
Protocol, 57
Protocol disk, 8
Storing, 57
Dispense, 185
Dispense custom command string, 245
Dispense fast command string, 245
Dispense slow command string, 245
Dispenser IN/OUT, 185

D
D
Delayed start, 34
Interrupt, 76
Data
Datafiles, 49
Pause in transfer, 75

310

5 Index

Dispensing, 219
Dispensing modules, 220
Dispensing offset, 244
Dispensing tube ID, 244
Dispensing volume, 220
Dispensing volume max., 245
Display output, 119, 224
DPM
Counting, 61
Standardization, 63

Coding, 31
Filtermats, 273
Flags, 93
Foil - heatsealing, 275
Format
Microtitration plate, 83
Format command, 50
FUNC
Function codes, 29
Function codes, 29

G
E

GenTerm, 152, 161, 238

Changing parameters, 162
Exit to MS-DOS, 163
Installation, 161
Printer connection, 163
Restrictions, 163
Result files, 163
Special features, 162
Start-up, 161
Trouble shooting, 163
GLP protocol, 111, 215, 253
Good Laboratory Practice, 111, 215, 253
Goto pos, 186

E
Counting, 35
Easy DPM, 64, 66, 125, 229
Edit standard curve, 126, 230
Editing input data, 16, 188
Efficiency, 269
Absolute, 103, 207
Relative, 103, 207
Efficiency correction
Detector efficiency correction, 257
Efficiency normalization, 255
Elapsed time, 28
Electronics, 268
End time, 28
Energy calibration
DPM standardization, 63
Enter key, 16, 188
Error beep, 147, 233
Errors, 69
EXCL, 130
External computer
Port 2, 49
External computer
Sending data to, 49
External output, 120, 145, 224

H
H
Help, 15, 73, 187
Half-life, 71
Half-life correction, 122, 129, 226, 258
Handshaking, 149, 235
Help, 15, 73, 187
System, 147, 233
Hiding text, 112, 216
High efficiency, 115, 219, 255
High voltage, 250
HiSafe, 88
How to control MicroBeta, 15, 187

Falcon plate, 123, 227

Faults, 252
Fibre optics, 251
Cleaning, 253
File output, 120, 145, 224
Filling wells, 88
Filtermat

Id label
Fixing, 29
ID label, 253
ID products, 275
ID support plate, 29

311

5 Index

ID system, 268
Info, 73
Init, 184
Initialize command string, 245
Injector, 242
Counting protocol parameters, 306
Initializing, 306
Installation, 301
Priming, 306
Test volumes and alignment, 306
Injector channels, 183
Injector module
Parameter setting, 208
Injector 1, 244
Injector assembly, 251, 266
Injector module, 183, 201, 243, 251
Injector setup, 183
Clear conveyor, 186
Dispense, 185
Dispenser IN/OUT, 185
Goto pos, 186
Init, 184
Next pos, 186
Off, 186
Prime, 184
Status, 186
Suckback, 185
Tip, 184
Wash, 185
Input file, 51
Inserts, 275
Installation, 287
Instrument description, 249
Interrupt, 75
Inverted plate, 117, 222
ISEP
Isotope spectrum end point, 258
Isotope, 115, 219
Activity setting (norm), 124, 228
Activity setting (stand), 125, 229
Spectrum end point, 258
Isotope 2 activity, 125, 229

L
L, 69
Labels - number of, 115, 218
LAN
Local area network, 151, 237
LCPS out of range monitor, 93
Lid, 75
Loading cassettes, 77
Local area network, 151, 237
Long output mode, 135
Low background, 115, 219
Luminescence
Coloured sealing tape, 201, 251
Counting, 79, 201
Flash, 201
Flash normalization, 202
Glow, 201
Glow normalization, 201
Normalization, 79
PMT selection, 115, 219
Units, 201
Luminescence assays
Dual luminescence, 251
Flash, 251
Glow, 250
Kinetic measurements, 251

M
M, 59
Mains voltage, 289
Mask adapter, 185, 243, 251, 253, 266
Robotic loading interface, 231
Mask Adapter
Parameter, 207
MAX, 94
Max. volumes in well, 88
Maximum plate time, 116
Maximum Well Volume, 245
Mean value, 261
Chi-square test, 263
Coefficient of variation, 262
Standard error, 262
Theoretical CV, 262
Theoretical error, 262
MeltiLex A, 274
MicroBeta JET
Background sample, 211

J
Jammed transportation system
Loading cassettes, 78

312

5 Index

MicroBeta JET
Background sample, 207
Injector system, 272
Serial port 3 parameters, 236
Microprocessor, 252
Microtitration plate, 83
Micro-volume LSC, 87
Mixing samples, 87
Monitors, 93
Out of range DPM, 93
Out of range LCPS, 93
Statistics, 93
MS-DOS, 156, 242
Definition, 159
MultiCalc, 95
Communication protocol, 101
Exit to MS-DOS, 102
Operation, 33
Printer connection, 102
Start up with, 291
Multichannel pump, 244
Multidrop dispenser, 89

Counting, 34, 35
Off, 75, 186
Operating conditions, 265
Out of range monitor
DPM, 93, 130
LCPS, 93, 130
Output
Display, external or file, 120, 224

P
ParaLux, 61, 63, 67, 255
ParaLux counting, 115, 219
Parameters in a protocol, 114, 218
Parity, 148, 235
Parking detectors, 300
Password, 122, 155, 156, 227, 241, 246
Pause
Data transfer, 75
Printing, 75
Performance, 268
Plastic bags, 274
Plate coding (MultiCalc), 100
Plate format
Microtitration plate, 83
Plate ID reader, 32, 268
PMT use, 115, 219
Polystyrene Safe, 274
Port 1, 49
Port 2, 49
Ports, 148, 234
Positions, 74
Power, 265
Power failure, 69
Power supply fan, 253
PREC, 94
Precision, 37, 116, 197, 221
Preset isotope protocols, 104, 209
Prime, 184
Prime command string, 245
Print through terminal, 101, 154, 240
Printer, 49, 150, 236
Connections, 298
Output, 117, 145, 222
Port 3, 49
Type, 298
Printing
Pause in printing, 75
Printout

N
N
Counting, 35
Next pos, 186
Next row, 75
No. of isotope standards (stand.), 125, 229
NORM, 107, 206, 214
Function codes, 29
Normalization, 103, 207, 255
Absolute or relative, 124, 228
And MultiCalc, 100
Protocol, 115, 218
Protocol parameters, 123, 227
With injection, 207
Without injection, 207
Normalization/standardization
Parameters, 205, 214
Number of channels, 244
Number of injectors, 244
Number of lines on screen, 153, 239
Nxx, 107, 206, 214

O
O, 77

313

5 Index

Changing field format, 132

Constant text, 133
Default formats, 136
Defining new fields, 133
Items, 127
Program failure, 69
Protocol name, 114, 218
Protocol number, 30
Protocols, 111, 215
Copying, 114, 218
Creating, 112, 216
Deleting, 114, 218
Editing, 112, 216
Listing, 111, 215
Parameters, 114, 218
Protecting, 114, 218
Types, 111, 215

S
Safety, 141
Sample plates, 272
Sample type, 125, 229
Sample volume, 220
Scintillation proximity assay, 89
ScintiStrip, 273, 274
Send result files to terminal, 153, 239
Setup mode, 156, 246
Shelf number, 34
Short output mode, 135
SPA, 89
SPA default curve fit method, 125, 230
Special plate, 122, 227
Specifications, 265
Spectrum indexes, 258
Spectrum quench index, 258
Spectrum quench parameter, 61
SQP(I), 61
Stability, 270
Standard curve fit selection (stand.), 125, 230
Standard deviation
Observed, 262
Theoretical, 262
Standardization, 63, 259
Standardization and MultiCalc, 100
Standardization protocol, 115, 218
Parameters, 124, 229
Standby power supply, 265
Start up
ParaLux, 293
Start up, 8, 290
Detector usage, 292
Plate ID reader, 293
Shelf usage, 292
Troubleshooting, 292
Statistical calculations, 261
Statistical deviation values, 129
Statistics, 143
Restrictions, 143
When available, 143
Statistics monitor, 93, 130, 261
Status, 186
Status display, 147, 233
STD
Function codes, 29
Stop, 75
STOP

Q
Quench series preparation, 88
Quenching, 63
Quick view, 116
Quick view, 115, 218
Quit, 156, 245

R
Radiation
Window, 179
Raise detector, 243
Repeat counting order, 38
With injection, 199
Without injection, 198
Repeats, 38, 198
Replicates, 38, 198
Results, 127
File, 51
Storing, 49
Return key, 16, 188
Robotic loading, 139, 231
Plate orientation, 117, 222
Robotic loading interface, 252
Routine maintenance, 253
RS-232C, 148, 234
RS232C protocol, 300

314

5 Index

Function codes, 29
Stop bits, 149, 235
Stop cassette
Cassettes, 30
Stop command string, 245
Storing results, 49
Strip plate, 43, 124, 228
Suckback, 185
Suck-back command string, 245
SuperMix, 87
Syringe volume, 244
System, 74, 147, 233
System parameter setting, 296

Exit to MS-DOS, 167

Installation, 165
Printer connection, 167
Restrictions, 167
Result files, 167
Special features, 167
Start up with, 290
Start-up, 166
Trouble shooting, 167
UltroTerm start up
Start up, 291
Usage, 74
Use injector, 219
Use mask adapter, 219
UTMAC, 171
Disk full, 172
Exit, 172
Installation, 171
Parameter changing, 171
Printer connection, 172
Result files, 172
Special features, 172
Start up, 171
Troubleshooting, 172

T
Tapes, 275
Technology, 95
Terminal, 152, 238
Terminal emulation, 159
Installation, 296
MultiCalc, 96
Terminal emulators, 159
Exit, 160
Startup, 160
Temporary exit, 160
Terminal, 159
What they are, 159
Terminal operation, 33
Terminal PC, 49
Port 1, 49
Terminal settings - exiting, 154, 240
Termination flag, 94, 130
Test function, 294
Test level, 294
Thermal background, 106, 210
Time, 27
Counting time, 28
Elapsed time, 28
End time, 28
Tip, 184
Tip init command string, 245
Total count rate, 177
Transportation system, 249, 266

V
Version, 148, 234
VT terminal start up
Start up, 291
VT100, 153, 239
VT52, 169
VT52, 96, 153, 169, 239
Installation, 169
Printer connection, 170
Restrictions, 169
Result files, 169
Special features, 169
Troubleshooting, 170

W
Wash, 185
Wash command string, 245
Window, 116, 179, 219
Settings, 179
Wrong key pressed, 69

U
UltroTerm, 152, 165, 238
Changing parameters, 166

315

5 Index

Zero date
Half-life, 71
Zero time
Half-life, 72

Z
Z
DPM standardization, 65

316