SPECTRAmax® PLUS

Microplate Spectrophotometer
Operator’s Manual

Molecular Devices Corporation

1311 Orleans Drive
Sunnyvale, California 94089

Part # 0112-0039
Rev. A
Molecular Devices Corporation
SPECTRAmax® PLUS Operator’s Manual
Copyright
 Copyright 1997, Molecular Devices Corporation. All rights reserved. No part of this publication may
be reproduced, transmitted, transcribed, stored in a retrieval system, or translated into any language
or computer language, in any form or by any means, electronic, mechanical, magnetic, optical, chem-
ical, manual, or otherwise, without the prior written permission of Molecular Devices Corporation, 1311
Orleans Drive, Sunnyvale, California, 94089, United States of America.
Patents
The SPECTRAmax PLUS and methods are protected by the following patents:
U.S. Patents 4,968,148 and 5,112,134; Canadian Patent 1,305,334.
Other U.S. and International patents pending.
Trademarks
SPECTRAplate, SPECTRAstrip, PathCheck and Automix are trademarks and SPECTRAmax and
SOFTmax are registered trademarks of Molecular Devices Corporation.
Apple and Macintosh are registered trademarks of Apple Computer, Incorporated.
All other company and product names are trademarks or registered trademarks of their respective
owners.
Disclaimer
Molecular Devices Corporation reserves the right to change its products and services at any time to
incorporate technological developments. This manual is subject to change without notice.
Although this manual has been prepared with every precaution to ensure accuracy, Molecular Devices
Corporation assumes no liability for any errors or omissions, nor for any damages resulting from the
application or use of this information.
Molecular Devices Corporation Instrument Warranty
Molecular Devices Corporation warrants this product against defects in material or workmanship as
follows:
1) All parts of the SPECTRAmax PLUS Microplate Spectrophotometer are warranted for a period of
one (1) year from the original date of delivery.
2) All labor charges to repair the product for a period of one (1) year from the original date of delivery
will be paid by Molecular Devices Corporation.
3) This warranty covers the SPECTRAmax PLUS instrument only, and does not extend to any com-
puter, printer, software, reagents, or disposables used with the instrument.

ii SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Labor and Parts
To obtain warranty service during the applicable warranty period, you must take the product or deliver
the product properly packaged in the original shipping materials and carton to an authorized Molecular
Devices Corporation service facility. You must call or write to the nearest Molecular Devices Corpora-
tion service facility to schedule warranty service. You may call Molecular Devices Corporation at the
telephone number or address below to locate the nearest service facility. You must schedule warranty
service prior to bringing or shipping the product for servicing. At the time of requesting warranty ser-
vice, you must present proof of purchase documentation which includes the date of purchase and Mo-
lecular Devices Corporation must have the enclosed Warranty Registration form completed, signed
and returned by you within ten (10) working days of the date of delivery.
This warranty only covers defects arising under normal usage and does not cover malfunctions of fail-
ures resulting from misuse, abuse, neglect, alteration, modification, or repairs by other than an autho-
rized Molecular Devices Corporation service facility.
Repair or replacement as provided under this warranty is the exclusive remedy of the purchaser (the
“Buyer”). Molecular Devices Corporation (the “Seller”) shall not be liable for any incidental or
consequential damages for breach of any express or implied warranty on this product, except
to the extent required by applicable law. The Seller specifically excludes all express and im-
plied warranties including without limitation any implied warranty that the products sold under
this agreement are merchantable or are fit for any particular purpose, except such warranties
expressly identified as warranties and set forth in the Seller’s current operating manual, cata-
log or written guarantee covering such product. The Seller also makes no warranty that the
products sold under this agreement are delivered free of the rightful claim of any third party by
way of patent infringement or the like. If the Buyer furnishes specifications to the Seller, the Buyer
agrees to hold the Seller harmless against any claim which arises out of compliance with the specifi-
cations.
Any description of the products contained in this agreement is for the sole purpose of identifying them.
Any such description is not part of the basis of the bargain, and does not constitute a warranty that the
products shall conform to that description. Any sample or model used in connection with this agree-
ment is for illustrative purposes only, is not part of the basis of the bargain, and is not to be construed
as a warranty that the products will conform to the sample or model. No affirmation of fact or promise
made by the Seller, whether or not in this agreement, shall constitute a warranty that the products will
conform to the affirmation or the promise.
For the name of the nearest authorized Molecular Devices Corporation service facility please contact
Molecular Devices at one of the following telephone numbers:
(408) 747-1700
(800) 635-5577 (U.S. and California)

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual iii

iv SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual
Contents

Figures and Table . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vi

Conventions Used in this Manual. . . . . . . . . . . . . . . . . . . . . . vii
Glossary of Terms. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Chapter 1 Instrument Description . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
General Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
PathCheck . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4
Component Description . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5
Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5
Control Panel Display . . . . . . . . . . . . . . . . . . . . . . . . . . 1-7
The Microplate Drawer . . . . . . . . . . . . . . . . . . . . . . . . . 1-8
The Cuvette Chamber . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9
The Back Panel. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-10
Functional Description . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-11
Stand-Alone Operation . . . . . . . . . . . . . . . . . . . . . . . . 1-11
Wavelength Selection . . . . . . . . . . . . . . . . . . . . . . . . . . 1-11
Temperature Regulation . . . . . . . . . . . . . . . . . . . . . . . . 1-11
Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-12
Printed Data Output . . . . . . . . . . . . . . . . . . . . . . . . . . 1-12
Computer Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-13
Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-16
Chapter 2 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
Installation Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
Installation Cautions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
Setting Up for Stand-Alone Use . . . . . . . . . . . . . . . . . . . . . . 2-4
Chapter 3 Stand-Alone Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1
Prepare for a Reading . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3
Turn the Instrument and Printer On . . . . . . . . . . . . . . . 3-3
Set the Temperature . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3
Select the Wavelength. . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3
Read the Cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4
Operation Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual v

 Chapter 4 Maintenance and Troubleshooting. . . . . . . . . . . . . . . . . . . 4-1
Technical Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Warnings and Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Changing the Fan Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Changing the Fuses. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-5
Error Codes and Probable Causes . . . . . . . . . . . . . . . . . . . . . 4-7
Error Messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-7
Opening the Drawer Manually . . . . . . . . . . . . . . . . . . . . . . . 4-9
Appendix A Printers and Cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-1
Compatible Printers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3
Cables. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3
Appendix B Accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1
Items Available for Use with the SPECTRAmax PLUS . . . . . B-3
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .I-1

Figures Figure 1.1: SPECTRAmax PLUS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3

Figure 1.2: Major Areas of the SPECTRAmax PLUS . . . . . . . . . . . . . . 1-5
Figure 1.3: Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5
Figure 1.4: Microplate Drawer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-8
Figure 1.5: Cuvette Chamber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9
Figure 1.6: Test Tube Cover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9
Figure 1.7: Components on the Back Panel of the
SPECTRAmax PLUS . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-10
Figure 1.8: Sample Printout from Thermal Printer . . . . . . . . . . . . . . . 1-13

Figure 2.1: View of Rear Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-4

Figure 4.1: Pry Open the Fuse Box Cover . . . . . . . . . . . . . . . . . . . . . . . 4-5

Figure 4.2: Removing the Fuse Box. . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-6
Figure 4.3: The Fuse Box and Holder with Fuses Removed . . . . . . . . . 4-6

Table Table 4.1: Error Codes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-7

vi SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Conventions The names of keys that appear on the SPECTRAmax PLUS control panel are
Used in this shown in boxed Helvetica type. Example: [Drawer].
Manual Italic and boldface type are used for emphasis. Examples: “Press carefully to
engage,” “Do not press down.”
NOTE: A note provides information that will help you properly execute an
action or procedure.
! CAUTION: Indicates an action or condition that could potentially damage
the instrument or one of its components or could result in loss of data.
WARNING: Indicates a situation that could result in potential injury to a
person working with the system.
BIOHAZARD: Indicates a condition involving potentially infectious biolog-
ical agents requiring that proper handling precautions be taken.

Glossary of Absorbance, A
Terms The amount of light absorbed by a solution. To measure absorbance accurately, it
is necessary to eliminate light scatter. In the absence of turbidity, absorbance =
optical density.
A = log (I0/I)
I0 = incident light
I = transmitted light
In this manual, we use the terms absorbance and optical density interchangeably.

Optical Density, OD
The amount of light passing through a sample to a detector relative to the total
amount of light available. Optical Density includes absorbance of the sample plus
light scatter from turbidity.

Transmittance, T
The ratio of transmitted light to the incident light.
T = I/I0
%T = 100 T

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual vii

viii SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual
Chapter 1 Instrument Description

Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
General Overview. . . . . . . . . . . . . . . . . . . . . . . . . 1-3
PathCheck . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4
Component Description . . . . . . . . . . . . . . . . . . . . . . . . . 1-5
Control Panel. . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5
Control Panel Display . . . . . . . . . . . . . . . . . . . . . 1-7
The Microplate Drawer . . . . . . . . . . . . . . . . . . . . 1-8
The Cuvette Chamber . . . . . . . . . . . . . . . . . . . . . 1-9
The Back Panel . . . . . . . . . . . . . . . . . . . . . . . . . 1-10
Functional Description . . . . . . . . . . . . . . . . . . . . . . . . . 1-11
Stand-Alone Operation . . . . . . . . . . . . . . . . . . . 1-11
Wavelength Selection . . . . . . . . . . . . . . . . . . . . . 1-11
Temperature Regulation . . . . . . . . . . . . . . . . . . 1-11
Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . 1-12
Printed Data Output . . . . . . . . . . . . . . . . . . . . . 1-12
Computer Control . . . . . . . . . . . . . . . . . . . . . . . 1-13
Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-16
Chapter 1

1-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

Introduction General Overview

The SPECTRAmax™ PLUS microplate spectrophotometer provides rapid and
sensitive UV/VIS measurements of a variety of analytes including specific pro-
teins, nucleic acids, and other molecules across a wide range of concentrations. It
measures the optical density (OD) of samples in a microplate or in a cuvette at a
selected wavelength for a single point in time (endpoint), over a specified period
of time (kinetic), or over a selected wavelength range (spectral scan).

Figure 1.1: The SPECTRAmax PLUS

The on-board microprocessor calculates and reports the absorbance (or % Trans-
mittance) for each well of a microplate. Data from multiple wavelengths can be
acquired during a single reading, if desired, and different calculations can be
made based on this data using SOFTmax PRO software, including the subtrac-
tion of blanks, use of standard curves, etc.
Typical applications include endpoint assays (quantitation of cytoproliferation by
MTT reduction, colorimetric protein assays, protein measurement at 280 nm,
DNA/RNA measurement at 260 nm, and endpoint ELISAs) and kinetic
measurements (enzyme studies, such as determination of the activity of enzymes
released from cells, and kinetic ELISAs).
All standard 96-well microplates, strip wells, and filter-bottom microplates can be
used in the SPECTRAmax PLUS. When reading at wavelengths below 340 nm,
special UV-transparent, disposable or quartz microplates and cuvettes that allow
transmission of the far UV spectra must be used.
With SOFTmax PRO, the contents of the wells in a microplate can be mixed
automatically by shaking before each read cycle, making it possible to perform
kinetic analysis of solid-phase, enzyme-mediated reactions (mixing is not critical
for liquid-phase reactions). The temperature of the microplate chamber can also
be regulated, if desired, from 4°C above ambient to 45°C.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-3

Chapter 1

An optional printer, for use with the cuvette chamber in a stand-alone mode, can
be purchased as an accessory.

PathCheck™
As predicted by the Beer Lambert law of light absorption, absorbance is propor-
tional to the distance that light travels through the sample—the longer the path-
length, the higher the absorbance.
Microplate readers use a vertical light path so the distance of the light through the
sample depends on the volume. This variable pathlength makes it difficult to per-
form extinction-based assays and confusing to compare results between micro-
plate readers and spectrophotometers.

Horizontal The standard pathlength of a cuvette is the

conventional basis for quantifying the unique
absorptivity properties of compounds in solution.
Horizontal
Quantitative analyses can be performed on the
Light Path basis of extinction coefficients, without standard
curves (e.g., protein at 280 nm, nucleic acid
estimates at 260 nm and NADH-based enzyme
assays). When using a cuvette, the pathlength is
known and is independent of sample volume, so
Cuvette absorbance is proportional to concentration.

In the microplate, pathlength is dependent on the

liquid volume, so absorbance is proportional to both
Vertical the concentration and the pathlength of the sample.
Standard curves are often used to determine analyte
concentrations in vertical-beam photometry of
Vertical Light Path unknowns, yet errors can still arise from pipetting the
samples and standards. The PathCheck feature of the
SPECTRAmax PLUS automatically determines the
pathlength of aqueous samples in the microplate and
normalizes the absorbance in each well to a
pathlength of 1 cm. This novel approach to
Microplate Wells correcting the microwell absorbance values is accurate
to within 2.5% of the values obtained directly in a 1-
cm cuvette.

Reference measurements made by reading the cuvette or using factory-stored val-

ues derived from deionized water can be used to normalize the OD data for
microplate wells.
This pathlength correction is accomplished only with the use of SOFTmax PRO
software. Stand-alone control of the instrument is limited to fixed-wavelength

1-4 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

reference measurements made with the cuvette. SOFTmax® PRO software from
Molecular Devices provides full instrument control and statistical data analysis.

Component The main components of the SPECTRAmax PLUS are:

Description • The control panel
• The microplate drawer
• The cuvette chamber
• The back panel (connections and power switch)

Back Panel

Control Panel Cuvette Chamber

Microplate
Drawer

Figure 1.2: Major areas of the SPECTRAmax PLUS

Control Panel

Figure 1.3: Control Panel

The control panel consists of an LCD and nine pressure-sensitive, color-coded
membrane keys which can be used to perform the stand-alone functionality of
the SPECTRAmax PLUS, including setting the temperature inside the micro-
plate drawer and cuvette chamber, setting the wavelength at which to read the
cuvette, toggling between displayed values as absorbance or percent transmission,

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-5

Chapter 1

and opening or closing the microplate drawer. Pressing a control panel key acti-
vates the function.
LCD
A 2-×-20-character liquid crystal display which shows the current instrument set-
tings.

Keys

TEMP
on/off (Incubator) Enables/disables the incubator function.

(Incubator) Allows you to enter a set point to regulate the mi-

croplate and cuvette chamber temperature. Pressing the up or
down arrow key scrolls up or down, starting at the previous
TEMP temperature setting (or the default of 37.0°C, if no setting had
been made). Pressing the up ( ▲) or down (▼) arrow once incre-
ments or decrements the temperature shown in the display by
0.1°C; pressing and holding either arrow increments or decre-
ments the temperature shown in the display by 1°C until it is
released. If you increment the setting to the highest limit (45°C)
and continue to press the up ( ▲) arrow, the display will not
change. If you decrement the setting to the lowest limit, 15°C,
and continue to press the down ( ▼) arrow, the display will not
change.
! CAUTION: If the incubator is disabled, pressing the [Temp On/Off] key
will enable the incubator.
Selects the wavelength that will be used for reading the cuvette
manually. Pressing the up or down arrow key scrolls up or
down through the available wavelengths, starting at the previ-
λ ous setting. Pressing the up (▲) or down (▼) arrow once incre-
ments or decrements the wavelength shown in the display by 1
nm; pressing and holding either arrow increments or decre-
ments the wavelength shown in the display by 10 nm until it is
released. If you increment the setting to the highest limit (1000
nm) and continue pressing the up (▲) arrow, the display
returns to the lowest possible setting (190 nm) and begins
incrementing from there. The inverse is true for decrementing
by pressing the down (▼) arrow.
NOTE: Setting the wavelength in SOFTmax PRO and reading using the soft-
ware will override the manual setting. The control panel will not show
the wavelength selected through SOFTmax PRO.

1-6 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

[REF] A reading of buffer, water, or air taken in the cuvette that is

used as I0 to calculate Absorbance or %Transmittance. If no
reference reading is taken, the SPECTRAmax PLUS will use
the I0 values stored in the NVRAM (non-volatile memory) of
the instrument.
Initiates the sample reading of the cuvette.
READ
CUVETTE

[%T/A] Toggle switch used to display cuvette data as percent trans-

mission or absorbance.
[Drawer] Opens or closes (toggles) the microplate drawer. Whether or
not the drawer will remain open depends on the incubator
setting. If the incubator is off, the drawer will remain open; if
the incubator is on, the drawer will close after approximately
10 seconds to assist in maintaining temperature control with-
in the microplate chamber.
NOTE: If the SPECTRAmax PLUS is being controlled by the computer soft-
ware, the [READ CUVETTE] and [REF] button will be inactive. If the
SPECTRAmax PLUS is idle, all keys on the control panel are active.
During a kinetic run, if the read interval is >30 seconds, pressing the
[Drawer] button will open the drawer. The instrument will automati-
cally close the microplate drawer for the next reading.

Control Panel Display

The left side of the display shows the temperature, both actual and set point, and
whether or not the temperature is at the set point (enunciator blinks if not at set
point). The middle of the display shows the wavelength. The right side of the dis-
play shows the data received from the reading as either absorbance or percent
transmission and indicates whether or not a reference measurement was made
(enunciator blinks if no reference reading was taken).
To change the contrast on the control panel, press [%T/A] and the [Temp] up
or down keys.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-7

Chapter 1

The Microplate Drawer

A1 Indicator

Figure 1.4: Microplate Drawer

The microplate drawer is located on the right side of the SPECTRAmax PLUS
and slides in and out of the reading chamber. Springs on two sides of the drawer
automatically position and hold a microplate in the proper position. The drawer
remains in the reading chamber during read cycles.
Microplate drawer operation varies, depending upon the incubator status. To
open the drawer, press the [Drawer] key. If the incubator is turned on, the
drawer will remain open for approximately ten seconds, after which a beeping
sound will alert you approximately two seconds before the drawer closes automat-
ically.
NOTE: Do not obstruct the movement of the drawer. If you must retrieve a
plate after an error condition or power outage and the drawer will not
open, it is possible to open it manually (see Chapter 4, “Maintenance
and Troubleshooting”).
Microplates
The SPECTRAmax PLUS can accommodate standard 96-well microplates, strip
wells, and filter-bottom microplates. When reading at wavelengths below
340 nm, special UV-transparent, disposable or quartz microplates allowing trans-
mission of the deep UV spectra must be used.
Not all manufacturers’ microplates are the same with regard to design, materials,
or configuration. Temperature uniformity within the microplate may vary
depending on the type of microplate used.

1-8 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

The Cuvette Chamber

Figure 1.5: Cuvette Chamber

Located at the right front of the SPECTRAmax PLUS, the cuvette chamber has a
lid that lifts up, allowing you to insert or remove a cuvette. The chamber contains
springs that automatically position the cuvette in the proper alignment for a read-
ing. The cuvette door must be closed in order to initiate a reading.
Cuvettes
The SPECTRAmax PLUS can accommodate standard-height (45 mm), 1-cm
cuvettes and 12 x 75 mm test tubes when used with the test tube cover.

Figure 1.6: Test Tube Cover

Not all manufacturers’ cuvettes are the same with regard to design, materials, or
configuration. Temperature uniformity within the cuvette may vary depending
on the type of cuvette used.
Be sure to handle cuvettes on the frosted sides only. Place the cuvette into the
chamber so that the “reading” (clear) sides face left and right (see Figure 1.5).
NOTE: The SPECTRAmax PLUS will not accept the short (25-mm high)
micro cuvettes.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-9

Chapter 1

The Back Panel

Power Switch

Fuse Box Cover Power Cord

Receptacle
Printer (Centronics)
Computer (RS-232)
Label

Figure 1.7: Components on the Back Panel of the SPECTRAmax PLUS

The following components are located on the back panel of the SPECTRAmax
PLUS:
• Power switch—a rocker switch, labeled I/O (for on and off, respectively).
• Power cord receptacle—plug the power cord in here.
• Fuse box cover—cannot be opened while the power cord is plugged in. When
opened, it provides access to the fuse box containing two fuses that are required
for operation.
• Printer port (double-shielded, 25-pin parallel, for use in stand-alone opera-
tion)—plug the 25-pin end of the cable into this port; the other (Centronics)
end attaches to the port on the printer.
• Computer port (double-shielded 8-pin RS-232 serial, for use with an external
computer)—plug the 8-pin DIN serial cable connecter into this port; the other
end attaches to the serial (modem) port of the computer.
• Labels—provide information about the SPECTRAmax PLUS, such as line
voltage rating, cautionary information, serial number, etc. Record the serial
number shown on this label for use when contacting Molecular Devices Tech-
nical Services.

1-10 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

Functional Stand-Alone Operation

Description The paragraphs that follow describe functions that can be activated during stand-
alone operation.
When operating the SPECTRAmax PLUS as a stand-alone system, you can
obtain a fixed-wavelength reading of the cuvette. The wavelength is chosen by
pressing the up or down arrows beneath the display labeled λ. Temperature regu-
lation of the cuvette/microplate chamber can be enabled and set using the keys
on the left side of the control panel and actual temperature can be monitored in
the display. To obtain a reading of the cuvette, press either the [REF] key (for a
reference reading) or the [READ CUVETTE] key (for a sample reading). The
resulting data can be displayed as absorbance or percent transmission by toggling
the [%T/A] key. You can also open or close the drawer using the [Drawer] key
on the right of the control panel.
NOTE: Keep cuvette door closed when reading a cuvette.

Wavelength Selection
The display on the control panel shows the currently selected measurement wave-
length (λ). You can change the wavelength by pressing the up or down arrows
until the display shows the desired wavelength. The wavelength will wrap around
to the top or bottom when the last selectable wavelength is reached.

Temperature Regulation
The SPECTRAmax PLUS has been designed to regulate the temperature of the
microplate chamber from 4°C above ambient to 45°C. Upon power up, when
the incubator is off, the temperature in the SPECTRAmax PLUS microplate
chamber is ambient and isothermal. Pressing the incubator [TEMP On/Off] key
will cause the SPECTRAmax to begin warming the microplate/cuvette chamber.
The temperature set point defaults to 37.0°C at start-up.
NOTE: Accuracy of the temperature set point is only guaranteed if the set point
is at least 4°C above ambient. If the temperature set point is lower than
the ambient temperature, the chamber temperature will remain at ambi-
ent. Temperature regulation is controlled by heaters only and, therefore,
cannot cool the temperature to a setting lower than ambient. Addition-
ally, the highest setting (45°C) can be achieved only if the ambient tem-
perature is >20°C.
You can change the temperature set point by pressing the up (▲) or the down
(▼) arrow keys above and below the word TEMP until the desired set point is
shown in the display. Typically, the microplate chamber will reach 37.0°C in 15
to 30 minutes.
The microplate chamber temperature is maintained at the set point until you
press the incubator [TEMP On/Off] key again, turning temperature regulation
off. The microplate drawer will open and the temperature within the chamber
will begin returning to ambient.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-11

Chapter 1

NOTE: Should you turn the incubator back on after a momentary shutdown,
allow about ten minutes after reaching temperature for the control algo-
rithm to fully stabilize the microplate chamber temperature.
Temperature regulation and control of the microplate/cuvette chamber are
achieved through electric heaters, a fan, efficient insulation, and temperature sen-
sors. The heaters are located in the chamber which is insulated to maintain the
temperature set point. The sensors, also mounted inside the chamber, measure
the air temperature. The temperature feedback closed-loop control algorithms
compare the measured air temperature inside the chamber against the tempera-
ture set point, and use the difference to calculate the heating cycles. This tech-
nique results in accurate, precise control of the chamber temperature with a
temperature variation of the air inside the chamber of less than 0.3°C. (The tem-
perature uniformity within the microplate or cuvette will depend upon the
design, materials, and/or configuration of that component.)

Data Collection
The SPECTRAmax PLUS stores only the most recent cuvette reading in a buffer
memory.
! CAUTION: Data in the buffer memory is lost when power to the
SPECTRAmax PLUS is turned off. This applies even to short power out-
ages. Do not turn the instrument off while important data remains in the
buffer memory.

Printed Data Output

An optional printer, designed to sit on top of the instrument, can be purchased
for use with the SPECTRAmax PLUS.
During stand-alone operation, results are automatically printed as soon as a
cuvette has been read. A new cuvette can be loaded into the SPECTRAmax
PLUS while the results from the first reading are being printed.

1-12 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

Figure 1.8: Sample Printout from Thermal Printer

NOTE: If you have performed a REF reading, the REF values will not be shown
as raw OD values, but are used to calculate subsequent sample reads
which are shown on the printout and display.

Computer Control
The SPECTRAmax PLUS is equipped with an 8-pin DIN RS-232 serial port
through which a computer can communicate with and control the instrument.
SOFTmax PRO®
Molecular Devices’ SOFTmax PRO software is a highly integrated program that
can be used to control and collect data from the SPECTRAmax PLUS. SOFT-
max PRO is easy to use, yet is powerful and flexible, and makes it possible to
access the full capabilities of the SPECTRAmax PLUS.
SOFTmax PRO allows you to:
• Read microplates for endpoint, kinetic, or spectral scans
- Use up to six wavelengths for endpoint and kinetic readings
- Perform Absorbance or %Transmittance readings in the 190- to 1000-nm
range
- Extend kinetic run times up to 99 hours

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-13

Chapter 1

- Select your own read intervals for kinetic runs

- Specify the duration for Automix before and between readings
- Read the whole plate or a subset of microplate strips
• Read cuvettes for endpoint, kinetic, or spectral scans
- Use up to six wavelengths for endpoint and kinetic readings
- Perform spectral scans from 190 to 1000 nm
- Extend kinetic run time up to 99 hours
- Select your own read intervals for kinetic runs
- Read a reference sample to be used with a single cuvette or a series of cuvettes
• Use PathCheck to normalize the absorbance readings in each microplate well to
a 1-cm pathlength
• Design microplate templates and cuvette templates to simplify data reduction
- Identify groups of wells or cuvettes with labels of your choice
- Identify individual wells or cuvettes with unique names
- Blank the entire plate, groups, and/or individual wells
• Save instrument settings, template formats, and data analysis parameters for
both microplates and cuvettes as assay protocol files and recall them for later
use
- Rapid instrument and analysis set up for repeated microplate or cuvette assays
- Uniform analysis for equivalent microplates and cuvettes
• Turn the incubator on or off to control the temperature in the microplate
drawer and cuvette port
• Acquire data from the SPECTRAmax PLUS
- Save data files for in-depth analysis at a later time
- Save multiple microplates and cuvettes with individual template and data
analysis parameters in one or more experiments in a single data file
- Pre-read microplates
- Analyze kinetic and spectrum data as it is collected
• Use the Automix function to shake the microplate at preset intervals, thereby
mixing the contents of each well (highly recommended for ELISAs and other
solid-phase, enzyme-mediated reactions)
• Display data on screen
- Raw values, reduced number, or raw values with reduced number
- Raw microplate data in a microplate format
- Ranged data as integers between 0 and 9 in a microplate format

1-14 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

- Threshold data as being above, below, or between set limits in a microplate

format
- Gray scale data in seven shades of gray corresponding to high and low limits
in a microplate format
- Kinetic or Spectrum plots of all 96 microplate wells
- Enlarge the display of individual well plots and overlay multiple well plots
- Multiple cuvette data as raw, ranged, threshold, gray scale, kinetic, or spectral
scans
• Perform data analysis using SOFTmax PRO features
- Calculate maximum kinetic rates on non-linear data
- Assign plate, group, or sample blanks
- Perform pathlength correction based on 1-cm cuvette readings
- Customize data analysis for each group in the template
- Create graphs with multiple plots
- Pick from nine curve-fitting routines
- Analyze unknown samples against a standard curve
- Analyze and compare data within a plate, between plates, and between exper-
iments
• Multiple print formats
- Print all or individual sections of the data file
- Define and print a report containing only selected sections
- Customize the order of data file sections
• Export data in tab-delimited ASCII format for use with Excel or other database
programs or as JCAMP-DX format for use with spectral analysis software
For a complete description of the features of SOFTmax PRO, refer to the SOFT-
max PRO User’s Manual.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-15

Chapter 1

Specifications Thermal specifications for microplates used in the SPECTRAmax PLUS apply to
flat-bottom microplates with isolated wells. All other microplate specifications
apply to standard 96-well polystyrene flat-bottom microplates. Performance spec-
ifications for cuvette readings apply only to aqueous solutions having solute molal
concentrations less than 0.4 M. When pathlength compensation is applied to
microplate absorbance measurements, agreement with cuvette absorbance mea-
surements for the same solution requires that the solution volume in the micro-
plate well be between 100 µL and 300 µL.
NOTE: Technical specifications are subject to change without notice.

Photometric Performance
Wavelength range 190–1000 nm
Wavelength selection Monochromator tunable in 1-nm increments
Wavelength bandwidth ≤ 2.0 nm FWHM (full width half maximum)
Wavelength accuracy ± 1.0 nm across wavelength range
Wavelength repeatability ± 0.2 nm across all optical channels
Photometric range -0.3 to 4.000 OD
Photometric resolution 0.001 OD
Photometric linearity (405 nm) 0.000 OD to 3.000 OD
Photometric accuracy (microplate) 0–2.0 OD: 190–1000 nm <± 1.0% and ± 0.006 OD
2.0–2.5 OD: 190–1000 nm <± 3.0% and ± 0.006 OD
Photometric accuracy (cuvette) 0–2.0 OD: 190–1000 nm <± 1.0% and ± 0.005 OD
2.0–2.5 OD: 190–1000 nm <± 3.0% and ± 0.005 OD
Photometric precision (repeatability) 0–2.0 OD: 190–1000 nm <± 1.0% and ± 0.003 OD
2.0–2.5 OD: 190–1000 nm <± 3.0% and ± 0.003 OD
Stray light ≤ 0.05% at 230 nm
Photometric stabilization Instantaneous
Photometric drift None—continuous referencing of monochromatic output
Calibration Automatic before first kinetic read and before every endpoint
reading
Optical alignment None required
Light source Xenon flash lamp (5 watts)
Average lamp lifetime 1 billion flashes
Illumination Top down (microplates); horizontal (cuvettes)
Photodetectors Silicon photodiode

1-16 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

Photometric Analysis Modes

Stand-Alone
• Single wavelength Absorbance or %Transmittance reading
of the cuvette (or test tube)
Using SOFTmax PRO
• Express data as Absorbance or %Transmittance
• Single wavelength reading of microplate and/or cuvette
• Multiple wavelength (up to six) reading of microplate and/
or cuvette
• Kinetic and kinetic graphics of microplate and/or cuvette
• Spectral scan (190–1000 nm) of microplate and/or cuvette

Measurement Time
Microplate
Read time (endpoint) • 96 wells in 12 seconds (single wavelength)
• 96 wells in 12* N seconds (N wavelengths)
Kinetic read intervals • 96 wells, 9-second minimum interval between readings
(single wavelength)
• 1 column, 2-second minimum interval between readings
(single wavelength)
Cuvette
Read time (endpoint) • 1 second (single wavelength)
Kinetic read intervals • 2-second minimum interval between readings
(single wavelength)

Scan Speed
Cuvette: Normal scan 60*K nm/min (K = wavelength interval)
Cuvette: Speed scan 200*K nm/min
Microplate: Normal scan 33*K nm/min (8-well strip)
Microplate: Speed scan 135*K nm/min (8-well strip)

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-17

Chapter 1

Temperature Regulation
Reading chamber Isothermal when temperature regulation is not enabled
Range 4°C above ambient to 45°C when temperature regulation en-
abled. The ambient temperature must be >20°C to achieve
temperature regulation at 45°C.
Resolution ± 0.1°C
Accuracy ± 1.0°C for microplate and cuvette chamber
Well-to-well uniformity at equilibrium ± 0.5°C at 37°C
Chamber warm-up time 15-30 minutes (measured on air) after initiation of tempera-
ture regulation
Temperature regulation 4 sensors
Variation < 0.3°C (regulated)
Temperature regulation diagnostics Temperature regulation system is continuously monitored
and updated
Evaporation Plate lid required to minimize evaporative cooling
Recommended microplate Flat-bottom microplates with isolated wells and lid

Automix with SOFTmax PRO

Plate mixing modes Selectable: off, once prior to any reading, and once prior to
and between kinetic readings
Plate mixing duration Selectable: 1 to 999 seconds (three-second default)

Compatibility
Microplate Standard and half-area 96-well flat-bottomed microplates
(0.3 mL). Polystyrene plates for wavelengths above 340 nm;
UV transparent plates above 220 nm; quartz plates above 190
nm.
Cuvette Standard height (45 mm) status cells with 10 mm pathlength
(12.5 mm x 12.5 mm outside) with minimum inside width
of 4 mm (typical for 3 mL volume cells).
Test Tubes 12 x 75 mm test tubes can be used in the cuvette chamber
with the test tube cover.

1-18 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Instrument Description

General Instrument
Display 2-×-20-character backlit LCD
Operating panel 9-key membrane keypad
Self-diagnosis Continuous on-board diagnostics
Spill control Drawer mechanism/reading chamber assembly is protected
from accidental spillage by drainage ports
Computer interface 8-pin DIN RS-232 serial (double shielding required)
Printer interface Parallel 25-pin to Centronics (double shielding required)
Microplates supported All 96-well and strip-well microplates, including lids

Environmental
Operating temperature 15 to 40°C
Operating humidity 0 to 85%, non-condensing
Storage temperature -20 to 65°C

Physical
Size (h × w × d) 8.6 in. (22 cm) × 22.8 in. (58 cm) × 15 in. (38 cm)
Weight 30 lb (13.6 kg)
Power consumption < 250 watts
Line voltage 90-250 VAC, auto-ranging
Line frequency 50/60 Hz

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 1-19

Chapter 1

1-20 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Chapter 2 Installation

Installation Warnings . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3

Installation Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
Setting Up for Stand-Alone Use . . . . . . . . . . . . . . . . . . . . 2-4
Chapter 2

2-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Installation

Installation 1) Always make sure the power switch on the instrument is in the OFF position
Warnings and remove the power cord from the back of the instrument prior to any instal-
lation or relocation of the instrument.
2) Do not operate the instrument in an environment where potentially damag-
ing liquids or gases are present.

Installation 1) Do not touch or loosen any screws or parts other than those specifically desig-
Cautions nated in the instructions. Doing so might cause misalignment and will void the
instrument warranty.

Unpacking The SPECTRAmax PLUS is packed in a specially designed carton. Please retain
the carton and the packing materials. If the unit should need to be returned for
repair, you must use the original packing materials and carton for shipping. If the
carton has been damaged in transit, it is particularly important that you retain it
for inspection by the carrier in case there has also been damage to the instrument.
WARNING: The SPECTRAmax PLUS weighs approximately 30 lb
(13.6 kg) and should be lifted with care. It is recommended that two per-
sons lift the instrument together, taking the proper precautions to avoid
injury.
After examining the carton, place it on a flat surface in the upright position.
Open the top of the box and lift the SPECTRAmax PLUS, along with the pack-
ing materials around the ends, up and out of the shipping box. Remove the pack-
ing material from both ends of the instrument and set the instrument down
carefully.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 2-3

Chapter 2

Setting Up 1) Place the SPECTRAmax PLUS on a level surface, away from direct sunlight,
for Stand- dust, drafts, vibration, and moisture.
Alone Use
2) Turn the instrument around so that the back of the instrument is facing you
as shown in Figure 2.1.

Power Switch

Fuse Box Cover Power Cord

Receptacle
Printer (Centronics)
Computer (RS-232)
Label

Figure 2.1: View of Rear Panel

3) Insert the appropriate (Macintosh or PC) serial interface cable into the RS-
232 serial port on the SPECTRAmax PLUS. Connect the other end to the
serial port of the computer.
4) Insert the female end of the power cord into the power receptacle at the rear
of the SPECTRAmax PLUS. Connect the male end to a grounded power
outlet of the appropriate voltage. Molecular Devices recommends that you
use a surge protector between the power cord and the grounded power outlet.
5) If you are using a printer for reading cuvettes in stand-alone mode, locate the
printer port (25-pin parallel) on the rear panel of the SPECTRAmax PLUS.
Connect one end of the cable to this port and the connect the other (Cen-
tronics) end to the printer. Load paper into the printer according to the man-
ufacturer’s instructions and connect the printer’s power cord to the power
outlet.
6) Turn the SPECTRAmax PLUS around so that the control panel now faces
you. Be sure no cables run beneath the instrument. Leave at least three inches
between the back of the instrument and the nearest objects or surfaces to
ensure proper ventilation and cooling.
7) Remove the tape from the cuvette door.
8) Turn on the power to the SPECTRAmax PLUS, wait for the microplate
drawer to open, and remove the tape and protective covering from the drawer
subplate.

2-4 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Chapter 3 Stand-Alone Operation

Prepare for a Reading . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Turn the Instrument and Printer On . . . . . . . . . . 3-3
Set the Temperature. . . . . . . . . . . . . . . . . . . . . . . 3-3
Select the Wavelength . . . . . . . . . . . . . . . . . . . . . 3-3
Read the Cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4
Operation Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4
Chapter 3

3-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Stand-Alone Operation

This chapter contains operating information for the SPECTRAmax PLUS Microplate/
Cuvette Spectrophotometer. If you are an experienced user of this instrument, you can
turn to the Operation Overview on page 3-4 for a quick review of the operating steps.
NOTE: Only temperature selection and single-wavelength cuvette readings are
available in stand-alone mode through front panel control. To achieve
full use of this instrument, connect it to a computer and use SOFTmax
PRO software.

Prepare for a Turn the Instrument and Printer On

Reading The power switch for the SPECTRAmax PLUS is located on the back panel.
Press the rocker switch to the ON position. The instrument will automatically
perform diagnostic checks to ensure that it is functioning correctly. Turn the
printer on at the same time.

Set the Temperature

If elevated temperature within the cuvette or microplate chamber is required, you
should turn on the incubator first, allowing enough time for the temperature to
reach the set point before performing a reading. When you first turn the instru-
ment on, up to 30 minutes may be required for the temperature within the
chamber to reach the set point.
To enable the incubator, press the incubator [TEMP On/Off] key. The LCD dis-
play will update to show the temperature set point and the current chamber tem-
perature.
To change the temperature set point, press the up or down arrows above and
below TEMP until the desired temperature set point is shown in the display.
The microplate/cuvette chamber temperature will be maintained at the
set point until you disable temperature control by touching the incubator
[TEMP On/Off] key again. When the incubator is off, the drawer will open
and the temperature within the chamber will begin returning to ambient.
NOTE: Should you turn the incubator back on after a momentary shutdown,
allow about ten minutes for the control algorithm to fully stabilize the
chamber temperature.

Select the Wavelength

Press the up or down arrow keys above and below the λ to increment or decre-
ment the wavelength setting (1 nm increments).

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 3-3

Chapter 3

Read the After setting the temperature (if desired), and choosing the wavelength for the
Cuvette reading, insert the cuvette into the chamber, making sure that the clear sides are
to the left and right (facing the instrument). Do not touch the clear surfaces of
the cuvette. Make sure the cuvette is completely seated in the chamber and close
the cuvette door. If the cuvette contains a blank (typically this solvent contains
everything that the samples contain except for analyte), press the [REF] key to
acquire the reference reading from the cuvette. The SPECTRAmax PLUS will
automatically calibrate in less than two seconds, send a header for the reading to
the printer, close the microplate drawer (if it was open), and read the cuvette
according to the selected instrument settings. If the cuvette contains a sample,
touch the [READ CUVETTE] key to acquire the sample reading from the
cuvette. When reading is complete, remove the cuvette.

Operation The following steps provide a quick reminder of the basic operating procedures
Overview required to read a cuvette using the SPECTRAmax PLUS.

1) Turn on the power switch of the SPECTRAmax PLUS (located on the back
panel).
2) If you wish to regulate the temperature inside the microplate reading chamber,
touch the [TEMP On/Off] key to bring the chamber to the default temperature
of 37.0°C. The microplate drawer will close and the indicator on the right of the
LCD will flash until the set temperature is reached.
3) If the incubator is on, the LCD will show the current temperature along with the
temperature set point. To change the set point (to any setting from ambient +4°
to 45°C), press either the up or down arrow above or below TEMP.
4) Select the desired measurement wavelength by pressing the up or down arrow
near λ. Scroll up or down through the list of wavelengths shown in the LCD
using the up or down arrows until the desired measurement wavelength is high-
lighted.
5) Load the prepared cuvette into the chamber, being sure that the clear sides are left
and right (when facing the instrument). Press the [REF] or [READ CUVETTE]
key.

3-4 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Chapter 4 Maintenance and Troubleshooting

Technical Support . . . . . . . . . . . . . . . . . . . . . . . . 4-3

Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Warnings and Cautions . . . . . . . . . . . . . . . . . . . . 4-3
General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Cleaning the Fan Filter . . . . . . . . . . . . . . . . . . . . 4-4
Changing the Fuses . . . . . . . . . . . . . . . . . . . . . . . 4-5
Error Codes and Probable Causes. . . . . . . . . . . . . 4-7
Error Messages . . . . . . . . . . . . . . . . . . . . . . 4-7
Opening the Drawer Manually . . . . . . . . . . . . . . 4-9
Chapter 4

4-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Maintenance and Troubleshooting

This chapter lists maintenance procedures and troubleshooting information,

including error codes that may be seen, followed by their most likely causes and
remedies.

Technical Molecular Devices Corporation is a leading worldwide manufacturer and distrib-

Support utor of analytical instrumentation. We are committed to the quality of our prod-
ucts and to fully supporting our customers with the highest level of technical
service. In order to fully benefit from our technical services, please complete the
registration card and return it to the address printed on the card.
If you have any problems using the SPECTRAmax PLUS Microplate Spectro-
photometer that are not covered in this chapter, in the U.S., contact our Techni-
cal Services group at 1-800-635-5577; elsewhere contact your local
representative.

Warnings BIOHAZARD: It is your responsibility to decontaminate the instrument, as

and Cautions well as any accessories, before requesting service by Molecular Devices
representatives and before returning the instrument or any components
to Molecular Devices Corporation.
WARNING: All maintenance procedures described in this manual can be
safely performed by qualified personnel. Maintenance not covered in this
manual should be performed by a Molecular Devices representative.
WARNING: Removal of protective covers that are marked with the High
Voltage warning symbol shown below can result in a safety hazard.

WARNING: Always turn the instrument off and remove the power cord and
any computer/printer cables from the back prior to any maintenance or
installation operation
WARNING: Never perform any operation on the instrument in an environ-
ment where liquids or potentially damaging gases are present.
WARNING: Risk of electrical shock. Refer servicing to qualified personnel.
! CAUTION: Use of organic solvents (such as dichloromethane) may cause
harm to the optics in the SPECTRAmax PLUS. Extreme caution is
advised when using organic solvents. Always use a plate lid and avoid
placing a plate containing these materials in the reading chamber for pro-
longed periods of time. Damage caused by the use of incompatible or
aggressive solvents is NOT covered by the instrument warranty.
! CAUTION: Never touch any of the optic mirrors, filters, or cables or their
housing, or manifold. The optics are extremely delicate, and critical toe
of the instrument.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 4-3

Chapter 4

! CAUTION: Do not touch or loosen any screws or parts other than those spe-
cifically designated in the instructions. Doing so could cause misalign-
ment and possibly void warranty.

General Keep the drawer closed when the instrument is not in use. The drawer can be
opened by pressing the DRAWER button. Always close the drawer immediately
prior to switching the instrument off.

Cleaning BIOHAZARD: Wear gloves during any cleaning procedure that could
involve contact with either hazardous or biohazardous materials or flu-
ids.
Periodically, you should clean the outside surfaces of the SPECTRAmax PLUS
using a cloth or sponge that has been dampened with water. Do not use abrasive
cleaners. If required, clean the surfaces using a mild soap solution diluted with
water or a glass cleaner and then wipe with a damp cloth or sponge to remove any
residue. Do not spray cleaner onto the instrument.
If needed, clean the microplate drawer or cuvette chamber using a cloth or
sponge that has been dampened with water.
Should fluids spill in the drawer area (when the drawer is out) or into the cuvette
chamber (when the door is open), they will be directed to a tray at the bottom of
the instrument from which they will exit to the bench or counter beneath the
instrument. Wipe up any spills immediately. Clean only the exterior of the unit
(and the microplate drawer or inside the cuvette chamber if necessary). Never
clean the inside of the instrument. Do not allow excess water or other fluids to
drip inside the instrument.

Cleaning the The fan filter on the bottom of the instrument requires periodic cleaning. The
Fan Filter frequency of the cleaning depends on how dusty your particular lab is and could
range from once a month to once every six months.
1) Turn power to the instrument OFF and then remove the power cord and
cables from the back of the instrument.
2) Make sure no plate or cuvette is in the instrument. Tape the cuvette door
closed. Turn the instrument over so that it rests flat on the bench.
3) Pop the black fan cover off the remove the filter.
4) Clean the filter by blowing clean, canned air through it or by rinsing it—first
with water and then with alcohol—and allowing it to dry completely.
5) Place the clean, dry filter over the fan and replace the black cover.
6) Turn the instrument back over. Remove the tape from the cuvette door.
Reconnect the power cord and cables to the instrument.

4-4 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Maintenance and Troubleshooting

Changing the Fuses burn out occasionally and must be replaced. If the instrument does not
Fuses seem to be getting power after switching it on (the LCD shows no display), first
check to see whether the power cord is securely plugged in to a functioning power
outlet and to the receptacle at the rear of the SPECTRAmax PLUS. If power
failed while the SPECTRAmax PLUS was already on, check that the power cord
is not loose or disconnected and that power to the power outlet is functioning
properly. If these checks fail to remedy the loss of power, follow the steps listed
below to replace the fuses. Spare fuses (two U.S. and two metric) are shipped with
the instrument in the original carton. The U.S. and metric fuses are identical
except for physical size. They may be taped to the back of the SPECTRAmax
PLUS.
If you no longer have spare fuses, you may obtain new ones from Molecular
Devices (part numbers: 4601-0013, U.S., 4601-0014, metric) or from a local
hardware store. Make sure fuses are rated SLOWBLOW (U.S.: 4-amp time-
delay; Metric: 4-amp, 5 × 20 mm, time-delay).
1) Switch power to the instrument off and then remove the power cord from the
outlet and from the SPECTRAmax PLUS power cord receptacle.
2) Remove the printer cable and computer cable (if connected) from the back of
the SPECTRAmax PLUS.
3) Turn the instrument around for easy access to the rear panel.
4) On the left-hand side of the rear panel (viewed from the back) is the power
cord receptacle. As shown in Figure 4.1, insert a small, flat-blade screwdriver
into the slot behind the tongue at the right of the black plastic cover. Gently
pry the cover open. It will begin to slide forward.

Figure 4.1: Pry Open the Fuse Box Cover

5) Continue gently prying the fuse box forward until you can pull it free of
the instrument. When removed, the fuse assembly will appear as shown in
Figure 4.2.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 4-5

Chapter 4

Figure 4.2: Removing the Fuse Box

6) Once the fuse box is out, you will see a holder inside containing two fuses.
Pull the fuse holder out of the box (see Figure 4.3).
7) It is possible that only one of the fuses may have blown. Molecular Devices
recommends that you replace both fuses, however, to ensure continued
proper operation. Pull both fuses out of the holder and discard them.

Figure 4.3: The Fuse Box and Holder with Fuses Removed
8) Insert new SLOWBLOW-rated fuses into the fuse holder. Either end of the
fuse may be forward.
9) Insert the fuse holder into the fuse box, making sure that the fuses face
toward the right (toward the tongue on the cover) as you insert it. Slide the
fuse holder all the way into the box.
10) Insert the fuse box into the opening in the instrument, making sure that the
fuses are on the right side (toward the power receptacle). Press the fuse box
into place, making sure the cover snaps closed.
11) Reconnect the power cord to the instrument and to the wall outlet and
reconnect other cables previously disconnected.

4-6 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Maintenance and Troubleshooting

Error Codes If a problem occurs during operation that causes an unrecoverable error, the
and Probable instrument will stop and an error code number will be shown in the display on
the front panel. To correct the problem, call your local Molecular Devices repre-
Causes sentative for assistance.

Error Messages
The LCD will display Fatal Error codes when a situation arises that requires
attention. Any reading in progress will stop. Warning messages do not stop a read-
ing but are logged in the error buffer. Warning messages indicate a situation that
requires attention but is not sufficient to stop or prevent a reading. Examples of
situations that might cause warning messages are low memory, entries being out
of range, or operations that could result in loss of data. These messages are gener-
ally self explanatory. For assistance regarding warning messages, contact your
local Molecular Devices representative.
Error Code Classifications
Not all error messages are listed in this manual. The errors are grouped in rela-
tionship to possible causes as follows:

Error Code Numbers Possible Causes

100 -199 Errors possibly caused by unrecognized commands being sent from the computer
to the instrument.
200-299 Errors probably due to a main board failure or a error in the firmware code. Most
of these errors require the assistance of Technical Support.
300-399 Instrument errors due to either a main board failure or other system failure. Most
of these errors require the assistance of Technical Support.
400-499 Errors due to failure or improper initialization of the instruments non-volatile
memory (NVRAM). All of these errors require the assistance of Technical Sup-
port.

Some errors are considered fatal in that if they are detected during power up (the
instrument will abort the power up sequence and display “FATAL ERROR ###”
on the LCD panel). Check the following list to see if there is something that you
can do to change the condition of the instrument to prevent the fatal error (e.g.,
closing the cuvette door during the power up sequence will prevent error num-
bers 111, 219, 302, and 310). Leave the instrument on for about five minutes,
turn it off and then back on. If you continue to get the fatal error message on
power up, record the error message number and contact Molecular Devices Tech-
nical Support or your local representative for assistance.
NOTE: If the instrument is functioning normally when using SOFTmax PRO,
no errors should be in the buffer (except error number 100).

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 4-7

Chapter 4

Table 4.1: Error Codes, Messages, and Notes about the Errors

Error
Error Message Notes
Code

100 command not found Command string was not recognized by the instrument.
101 invalid argument Command Argument does not match specified parameters or
syntax.
102 invalid data Command argument is out of acceptable range.
103 too many arguments Too many arguments after a specific command.
104 not enough arguments Not enough arguments for a given command.
105 input line too long Too many characters in the input line.
106 command error, system busy Instrument could not perform the given command because it was
busy doing another task. Example: Request a wavelength while
the monochromator is in motion.
107 command error, measure- Instrument could not perform the command because a measure-
ment in progress ment was in progress.
108 command error, no data A request to the instrument to transfer data when there’s no data
available in the buffer.
109 data buffer full To many data sets in the buffer. Can be caused by setting up a
long kinetic run and disconnecting the computer or when Soft-
Max Pro is pre-empted by another application.
110 error buffer full More than 65 errors in the buffer.
111 stray light cuvette, door open Cuvette door open while doing a read. Note error’s # 219 and
302 and 310 can also be caused by this.
205 user generated bus error Result of improper use of certain commands.
209 receive buffer overflow Caused by an external device sending too much data over the
serial port and ignoring flow control.
210 serial port parity error Parity bit error detected with incoming serial data.
211 serial port overrun error Caused by host computer sending too much data and ignoring
the flow control signal.
219 Cuvette gain check fail Power up check of cuvette’s amplifier’s gain is out of tolerance.
Could be due to bad or disconnected pre-amp board or excessive
stray light or cuvette door being open on power up.

4-8 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Maintenance and Troubleshooting

Table 4.1: Error Codes, Messages, and Notes about the Errors

Error
Error Message Notes
Code

300 temp sensor faulty Unable to read a reasonable thermistor value. Thermistor is
faulty or disconnected, Main board problem, or ambient temper-
ature is out of range.
301 safe temperature limit A temperature of over 50 deg C was detected on one or more of
exceeded the 4 thermistors. Temperature will be shut off and remain off
until power up reset is completed successfully.
302 low light measurement Not enough light detected to make an accurate measurement. If
reading a cuvette, make sure the cuvette door is closed.
310 Signal level saturation During a cuvette read, make sure the cuvette door is closed. Dur-
ing a plate read, make sure nothing is blocking the door from
shutting completely.
317 LCD time-out LCD front panel is bad or disconnected.
400 - Errors in Non-Volatile Mem- Contact Molecular Devices Technical Support.
499 ory

Opening the If an error occurs while the drawer is closed and you need to open the drawer,
Drawer press the [Drawer] key. If the drawer does not open, turn power to the instru-
ment off and then on again.
Manually
If the drawer still remains closed, turn the power off and using your thumb nail,
locate the groove in the upper left side wall of the door. Open the door, and with
your index finger, pull the microplate drawer out of the instrument (do not force
the drawer) and remove the microplate. This action will not harm the instrument,
but should only be taken if the first two options have failed to open the drawer.
If you are still unable to open the drawer, contact Molecular Devices Technical
Support or your local representative for assistance.

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual 4-9

Chapter 4

4-10 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Appendix A Printers and Cables

Compatible Printers . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3

Cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3
Appendix A

A-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Printers and Cables

Compatible The SPECTRAmax PLUS can be used with an Epson-compatible strip printer.
Printers Compatibility may require changing the dip switch or pin settings on the printer
or insertion of an emulation cartridge. Please check your printer’s manual to
assess compatibility based on this criterion—some printers will not meet this
requirement.
Please contact your local Molecular Devices representative for information
regarding recommended printers.

Cables Molecular Devices recommends that you use high-quality, double-shielded cables
to connect the SPECTRAmax PLUS to other peripheral instruments, such as a
printer or computer. Choose cables that meet the following requirements:
Printer Cable (for Stand-alone operation only):
Centronics parallel, male 25D to Centronics.
Serial Interface Cable
(contact Molecular Devices for specific pin-out requirements)
Macintosh: Male DB8 to male DB8
IBM Compatible: Male DB8 to Female DB9 (custom cable made by
Molecular Devices)

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual A-3

Appendix A

A-4 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Appendix B Accessories

Items Available for Use with the

SPECTRAmax PLUS . . . . . . . . . . . . . . . . . . . . . . . . . . B-3
Appendix B

B-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Accessories

Items
Available for
Use with the
SPECTRA-
max PLUS
Part Number
Cable, RS-232, Macintosh to SPECTRAmax PLUS,
8-pin DIN to 8-pin DIN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0091
Cable, RS-232, PC to SPECTRAmax PLUS,
9-pin to 8-pin DIN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0149
Cable, parallel printer, 25-pin to Centronics . . . . . . . . . . . . . . . . . . . 9000-0003
Cuvette Calibration Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0161
Fuse, 4-amp Time Delay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4601-0013
Fuse, 4-amp (5 × 20 mm) Time Delay. . . . . . . . . . . . . . . . . . . . . . . . 4601-0014
Power Cord (US, Canada, Japan, Mexico, India) . . . . . . . . . . . . . . . . 4400-0002
Power Cord (Germany, France, Scandinavia, Italy, Korea) . . . . . . . . . 4400-0036
Power Cord (UK, Indonesia, Singapore, Malaysia). . . . . . . . . . . . . . . 4400-0037
Power Cord (Australia, Hong Kong, China) . . . . . . . . . . . . . . . . . . . 4400-0038
Printer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0189
SPECTRAplate—Disposable
UV-transparent disposable microplates (case of 50) . . . . . . . . . . . . . . R9013
SPECTRAplate—Quartz
UV-transparent quartz microplate . . . . . . . . . . . . . . . . . . . . . . . . . . . R8024
SPECTRAmax Mouse Pad. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0133
SPECTRAtest PLUS Validation Package . . . . . . . . . . . . . . . . . . . . . . 0200-2405
Test tube cover. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2300-0277

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual B-3

Appendix B

B-4 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Index

A I
Accessories B-1 Incubator 1-8, 1-11, 3-3
Installation 2-3
B Cautions and Warnings 2-3
Back Panel 1-10
Bandwidth 1-16 K
Keys (control panel) 1-5, 1-6, 3-4
C
Cables 2-4, A-3 L
connecting 2-4 Labels 1-10
serial 1-10 LCD (liquid crystal display) 1-6, 3-4, 4-7
Calibration 1-16, 1-17
Cleaning 4-4 M
Component description 1-5 Maintenance 4-1, 4-4
Computer control 1-13 Microplate 1-3
Connections 1-5, 1-10 96-well 1-8
cables 2-4, A-3 filter-bottom 1-8
printer 1-10, 2-4 strip wells 1-8
Control panel 1-5, 1-7 temperature uniformity 1-8
Cuvette 1-5, 1-9 Microplate drawer 1-5, 1-7, 1-8
closing 1-8
D incubator status 1-8
Data open manually 4-9
collection 1-12 Microplates 1-8
printed output 1-12
Drawer 1-5, 1-7, 1-8 P
closing 1-8 Pathlength correction 1-4
incubator status 1-8 Ports 2-4
open manually 4-9 printer 1-10
Drawer key 1-7, 1-8, 1-11 serial 1-10
Power cord 1-10, 2-4, B-3
E Power switch 1-5, 1-10, 3-3
Error messages 4-7 Print 1-12
Printed Data Output 1-12
F Printer 1-4, 1-12, 1-13, 2-4, 3-3, A-1
Fuses Printer port 1-10, 2-4
accessing 1-10
changing 4-4 R
Read mode 1-13, 3-4
Read the microplate 1-13, 1-14, 3-4
preparation 3-3
Reference reading 1-7, 3-4
Remote control 1-13

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual I-1

Index

S
Serial port 1-10
Set point, temperature 1-6, 1-7, 1-12, 3-3
SOFTmax PRO (software) 1-3, 1-5, 1-13
Specifications 1-16
Spills 4-4
Stand-alone operation 1-11, 2-1, 2-4, 3-1

T
Technical service 4-3
Temperature 1-11, 1-12, 3-4
equilibration 1-12, 3-3
incubator 1-12, 3-3
regulation 1-11
set point 1-6, 1-7, 1-12, 3-3
setting 3-3
stabilization 1-12

U
Unpacking 2-3

W
Warranty ii
Wavelength 1-6, 3-3, 3-4
range 1-16
selection 1-11

I-2 SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

 Contents v
Figures vi
Table vi
Conventions Used in this Manual vii
Glossary of Terms vii
Absorbance, A vii
Optical Density, OD vii
Transmittance, T vii
Introduction 1-3
General Overview 1-3
PathCheck™ 1-4
Component Description 1-5
Control Panel 1-5
LCD 1-6
Keys 1-6
Control Panel Display 1-7
The Microplate Drawer 1-8
Microplates 1-8
The Cuvette Chamber 1-9
Cuvettes 1-9
The Back Panel 1-10
Functional Description 1-11
Stand-Alone Operation 1-11
Wavelength Selection 1-11
Temperature Regulation 1-11
Data Collection 1-12
Printed Data Output 1-12
Computer Control 1-13
® 1-13
SOFTmax PRO
Specifications 1-16
Installation Warnings 2-3
Installation Cautions 2-3
Unpacking 2-3
Setting Up for Stand-Alone Use 2-4
Prepare for a Reading 3-3
Turn the Instrument and Printer On 3-3
Set the Temperature 3-3
Select the Wavelength 3-3
Read the Cuvette 3-4
Operation Overview 3-4
Technical Support 4-3
Warnings and Cautions 4-3
General 4-4
Cleaning 4-4
Changing the Fuses 4-4
Error Codes and Probable Causes 4-7
Error Messages 4-7

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual i

Error Code Classifications 4-7
4-9
Opening the Drawer Manually 4-9
Compatible Printers A-3
Cables A-3
Items Available for Use with the SPECTRA-max PLUS B-3

ii SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual

Figure 1.1 : The SPECTRAmax PLUS1-3
Figure 1.2 : Major areas of the SPECTRAmax PLUS1-5
Figure 1.3 : Control Panel1-5
Figure 1.4 : Microplate Drawer1-8
Figure 1.5 : Cuvette Chamber1-9
Figure 1.6 : Test Tube Cover1-9
Figure 1.7 : Components on the Back Panel of the SPECTRAmax PLUS1-10
Figure 1.8 : Sample Printout from Thermal Printer1-13
Figure 2.1 : View of Rear Panel2-4
Figure 4.1 : Pry Open the Fuse Box Cover4-5
Figure 4.2 : Removing the Fuse Box4-5
Figure 4.3 : The Fuse Box and Holder with Fuses Removed4-6

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual i

ii SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual
: Error Codes, Messages, and Notes about the Errors 4-8

SPECTRAmax PLUS Microplate Spectrophotometer Operator’s Manual i