ADAMASCIENCEANDTECHNOLOGYUNIVERSI

TY
SCHOOLOFAPPLIEDNATURALSCI
ENCE
PROGRAM OFAPPLI
EDBI
OLOGY

GeneralBi
ologyPr
act
ice(
Biol
1111)
Teachi
ngManual

Adama,
Ethi
opi
a

i
 Oct
ober2017

Pr
eface
Thi
s Labor
ator
y Manualwas pr
epar
ed f
ort
he cour
se ‘
Gener
alBi
ology Pr
act
ice
(
Biol
1111)
’whi
chi
sgi
vent
oal
lfr
eshmanpr
esci
encest
udent
sjoi
ningt
heSchoolof
Appl
i
edNat
uralSci
ence,
AdamaSci
enceandTechnol
ogyUni
ver
sit
y.Thecour
sehas10
l
abor
ator
yst
udi
eswi
thdi
ff
erentsessi
ons.Aut
hor
sfort
hismanualr
ecommendt
hatt
he
st
udent
sshoul
dfami
l
iar
izet
hemsel
vesbyr
eadi
ngt
heexper
iment
stobeper
for
medi
n
t
hemanual
.Ther
efor
e,t
heexper
iment
sshoul
dber
eadbef
orecomi
ngt
othel
abor
ator
y,
butt
hepr
act
icalexer
ciseshoul
dbedonei
nthel
abor
ator
ycl
assi
tsel
f.Thest
udent
s
shoul
dindi
vi
dual
l
ypr
epar
ear
epor
tandsubmi
tther
epor
tatt
heendofeachexper
iment
of
fcour
sewi
tht
headv
iceofani
nst
ruct
or.Thebackgr
oundgi
veni
neachl
abor
ator
y
st
udywi
l
lgi
vesuf
fi
cientt
heor
eti
cal
inf
ormat
ionaboutt
hel
ab.

i
i
 I
nst
ruct
iont
ost
udent
s

1.Comet
othel
abor
ator
yint
ime.
2.Al
way
sbr
ingy
ourl
abmanual
,separ
atenot
ebookf
ort
hecour
se,penci
l
,er
aser
,
andr
uler
.
3.I
fyoumi
ssal
abf
orr
easonsbey
ondy
ourcont
rol
,repor
tthecaset
othel
ab
r
esear
chassi
stanti
nti
me.Onl
yreasonabl
eexcusesshal
lbeaccept
edt
oar
range
amake-
upl
ab;
other
wise,
youwi
l
lrecei
vezer
ofort
hel
aby
oumi
ssed.
4.Mostl
abor
ator
yact
ivi
ti
esar
egr
oupwor
k.I
nsuchcasesal
lthemember
soft
he
gr
oupshoul
dpar
ti
cipat
eint
hegr
oupact
ivi
ti
es.
5.Youar
eresponsi
blef
oral
llabor
ator
yequi
pmentpr
ovi
dedf
ory
ouruse.I
fyou
obser
veanydamageony
ourequi
pment
,repor
tatt
hebegi
nni
ngoft
heper
iod,
ot
her
wisey
oubehel
dresponsi
ble.
6.Youi
ndi
vi
dual
l
ywr
it
eneatandr
eadabl
elabr
epor
tforeachpr
act
icalandsubmi
t
i
ttot
hei
nst
ruct
ori
nti
me.Thel
abr
epor
twr
it
ingf
ormati
sat
tachedatt
heendof
t
hismanual
(Appendi
x).

i
i
i
 TABLEOFCONTENT
CONTENTS PAGE

LABORATORYSTUDYNUMBER1

GENERALLABORATORYSAFETYRULES-
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LABORATORYSTUDYNUMBER2

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CROSCOPES-
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LABORATORYSTUDYNUMBER3

THECELLANDCELLSTRUCTURE-
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LABORATORYSTUDYNUMBER4

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FFUSI
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LABORATORYSTUDYNUMBER5

TESTI
NGFORBI
OLOGI
CALLYI
MPORTANTMOLECULES-
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LABORATORYSTUDYNUMBER6

BI
OCATALYSI
S–ENZYMES-
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LABORATORYSTUDYNUMBER7

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-54

LABORATORYSTUDYNUMBER8

i
v
MI
CROBI
OLOGI
CALLABORATORYTECHNI
QUES-
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59

LABORATORYSTUDYNUMBER9

CELLDI
VISI
ON-
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-66

LABORATORYSTUDYNUMBER10
COLLECTI
ONANDPRESERVATI
ONOFPLANTANDI
NSECTSPECI
MENS-
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69
REFERENCES--
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APPENDIX--
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87

v
 LABORATORYSTUDYNUMBER1

GENERALLABORATORYSAFETYRULES

1.
1.I
ntr
oduct
ion
Al
abor
ator
yisaf
aci
l
ityt
hatpr
ovi
descont
rol
l
edcondi
ti
onsi
nwhi
chsci
ent
if
icr
esear
ch,
exper
iment
s,andmeasur
ementi
sper
for
med.Thesci
ent
if
icmet
hodi
sapr
ocessf
or
exper
iment
ati
on t
hat i
s used t
o expl
ore obser
vat
ions and answer quest
ions
sci
ent
if
ical
l
y.Sci
ent
ist
suse t
hesci
ent
if
icmet
hod t
o sear
ch f
orcause and ef
fect
r
elat
ionshi
psi
nnat
ure.I
not
herwor
ds,t
heydesi
gnanexper
imentsot
hatchangest
o
onei
tem causesomet
hingel
set
ovar
yinapr
edi
ctabl
eway
.Justasi
tdoesf
ora
pr
ofessi
onalsci
ent
ist
,thesci
ent
if
icmet
hodwi
l
lhel
pyout
ofocusy
oursci
encef
air
pr
ojectquest
ion,
const
ructahy
pot
hesi
s,desi
gn,
execut
e,andev
aluat
eexper
iment
s.

Saf
etyr
ulesar
egui
del
i
nesdesi
gnedt
ohel
pandkeepy
ousaf
ewhenexper
iment
ing.
Someequi
pmentandchemi
cal
sinal
abor
ator
ycancauseser
ioushar
ms.I
tisal
way
s
wi
set
ofol
l
owal
ll
abor
ator
ysaf
etyr
ules.

Thef
oll
owi
ngl
abor
ator
ysaf
etyr
ulesar
easampl
eoft
hemostbasi
crul
est
hatshoul
d
bef
oll
owedwheni
nwor
kingi
nthel
abor
ator
y.Mostl
abor
ator
ieshav
ethesaf
etyr
ules
post
edi
nthel
abor
ator
yandt
hei
nst
ruct
or/
trai
nerwi
l
lmostl
i
kel
ygoov
ert
hem,
wit
hyou,
bef
orey
oubegi
nwor
king.

1.Bef
orey
ouent
eral
abor
ator
y,y
oushoul
dbepr
epar
edf
orknowl
edgeabl
eabout
anyl
abor
ator
yexer
cisest
hatar
etobeper
for
med.Thatmeansy
oushoul
dread
y
ourl
abor
ator
ymanual
toknowexact
lywhaty
ouwi
l
lbedoi
ng.
2. Uponent
eri
ngt
hel
abor
ator
y,pl
aceal
lbooks,backpacks,andl
argeper
sonal
i
temsont
he desi
gnat
edshel
ves.Donotcl
utt
ery
ourwor
kar
ea.
3.Locat
efi
reext
ingui
sher
,ey
ewash,f
ir
stai
dki
t,br
okengl
asscont
ainer
,andcl
ean
upmat
eri
alsf
orspi
l
ls.
4.I
ncaseoff
ir
e,uset
henear
estexi
ttoev
acuat
ethebui
l
ding/
room i
naqui
ck&
cal
m manner
.

1
 5.I
fcl
othi
ngshoul
dcat
chf
ir
e,smot
heri
twi
thabl
anketorcoat
.NEVERRUN!
!
6. Whenwor
kingi
nal
abor
ator
y,makesur
eyoukeepy
ourar
eaneatandor
gani
zed.
I
fyouhappent
ospi
l
lsomet
hing,askf
orassi
stancewhencl
eani
ngi
tup.Al
so
r
emembert
ocl
eany
ourwor
kar
eaandwashy
ourhandswheny
ouhav
efi
nished
t
heact
ivi
ty.
7.Remov
eort
iebackanyar
ti
cleofcl
othi
ngorj
ewel
ryt
hatcanhangdownand
t
ouchchemi
cal
sandf
lames.Youwi
l
lal
sowantt
owearpr
opershoest
hatcan
pr
otecty
ourf
eeti
ncasesomet
hingget
sbr
oken.Donotwearsandal
soranyt
ype
ofopen-
toedshoesi
nthel
abor
ator
y.
8.Wearey
epr
otect
ionwhenr
equest
edbyy
ouri
nst
ruct
or.
9. Repor
tal
lspi
l
ls,acci
dent
s,andi
njur
iest
oyouri
nst
ruct
or(
washwi
thwat
eror
asky
ourt
eacherf
orhel
p)
10.
Washski
nimmedi
atel
ywi
thsoapandwat
eri
fcont
ami
nat
edwi
thchemi
cal
sor
mi
croor
gani
sms.
11.Youmaybewor
kingwi
thgl
assorshar
pobj
ect
s,soy
oudon’
thandl
ethem
car
elessl
y.
12.
Remov
eaf
fect
edcl
othi
ngt
opr
eventr
eact
ionwi
thy
ourski
n.
13.
Biol
ogi
cal
wast
emustbedi
sposedofpr
oper
lyi
ntheBi
ohazar
dWast
eCont
ainer
.
14.
Keepf
lammabl
eli
qui
dsawayf
rom openf
lamesandheatsour
ces.
15.
Uponcompl
eti
onofl
abor
ator
yexer
cises,
placeal
lmat
eri
alsi
ndesi
gnat
edar
eas.
16.
Donotpourl
i
qui
dwast
esi
nthesi
nkunl
esst
oldt
odosobyy
ouri
nst
ruct
or
17.Washy
ourhandswi
thsoapandwat
erpr
iort
oleav
ingl
abor
ator
y.
18.
Donotr
emov
emat
eri
alsf
rom t
hel
abor
ator
y.

Ther
ear
esev
eralt
hingsi
nabi
ologyl
abor
ator
ythaty
oumustav
oidsuchas:
 Donott
ast
e/smel
lanychemi
cal
sorsubst
ancesy
ouar
ewor
kingwi
th
 Donotusey
ourmout
hforpi
pet
ti
ngsubst
ances

 Donothandl
ebr
okengl
asswi
thbar
ehands
 Donotpourchemi
cal
sdownt
hedr
ainwi
thoutper
missi
on
 Donotoper
atel
abor
ator
yequi
pmentwi
thoutper
missi
on

2
  Donotl
eav
eanyheat
edmat
eri
alsunat
tended
 Donotper
for
myourownexper
iment
sunl
essgi
venper
missi
on
 Donotpl
acef
lammabl
esubst
ancesnearheat
 Donoteat
,dr
inkorchewgum i
nthel
abor
ator
y

1.
2Somebasi
cequi
pmentusedi
nbi
ologyexper
iment
s

Thedai
l
yrout
ineofabi
ologi
sti
nvol
vest
heuseofbasi
cequi
pmenti
nthei
rbi
ology
exper
iment
s-suchasmi
croscopes,
testt
ubes,
beaker
s,andBunsenbur
ner
saswel
las
hi
gh-
techsci
ent
if
icequi
pmentandcomput
ers.Thi
sequi
pmenti
sthebar
e-bonebasi
cs
t
haty
ou’
dfi
ndi
nanyl
abor
ator
y.Thi
sequi
pmenti
snecessar
yfort
hebasi
cst
udi
esof
bi
ology
:vi
sual
i
zingcel
l
sandor
ganel
l
es,aswel
laspr
epar
ingsampl
esofcel
l
sorf
lui
ds
f
ort
est
ingorv
isual
i
zat
ion,
dissect
ingspeci
mens,
ormi
xi
ngchemi
cal
s.

1.Magni
fyi
nggl
ass(
handl
ens)-i
saconv
exl
ensmount
edi
naf
rame.I
tisusedt
o
pr
oduceamagni
fi
edi
mageofanobj
ect
2. Pet
riDi
sh-i
tisusedf
orhol
dingacul
tur
emedi
um uponwhi
chcel
l
s,bact
eri
a
andv
irusescanbegr
ownandst
udi
ed.Ot
herusesoft
hePet
ridi
sh:t
oteach
st
udent
saboutseedger
minat
ion,ast
hecl
eardi
shal
l
owst
heobser
vert
osee
ev
eryst
epofgr
owt
h,commonl
yusedf
ordi
ssect
ion,
becausei
tisi
deal
l
ysi
zedt
o
bepl
acedunderami
croscopeandcanal
sobeusedf
orbasi
cexper
iment
al
pur
posesl
i
ket
ranspor
ti
ngl
i
qui
dsi
nst
eri
l
econt
ainer
sordr
yingf
lui
dsf
orst
udy
.
3. Cov
ersl
ip-i
tisapr
otect
ort
hatgoesov
ert
issueswi
thmount
ingmedi
a,t
his
keepst
het
issueont
hesl
i
def
rom degr
adi
ng
4. Ther
momet
er-i
tisusedf
ormeasur
ingt
emper
atur
eort
emper
atur
egr
adi
ent
usi
ngav
ari
etyofdi
ff
erentpr
inci
ples
5. Testt
ube – i
sbasi
cal
l
ya nar
row gl
asst
ube used f
orst
ori
ng t
hingsand
per
for
mingsmal
lchemi
calr
eact
ionsi
nal
abor
ator
y;i
tisusedhol
d,mi
xorheat
smal
lquant
it
iesofsol
i
dorl
i
qui
dchemi
cal
s
6.Mi
cror
cent
ri
fugeHol
dsEppendor
f,ormi
crocent
ri
fuge,
tubest
hatcanhol
dabout
1.
5ml
ofl
i
qui
d
7.Mi
croscope-i
tisusedf
orv
iewi
ngobj
ect
sthatar
etoosmal
ltobeseenbyt
he
nakedey
es.

3
8. Funnel-i
tisusedt
ochannell
i
qui
dorf
ine-
grai
nedsubst
ancesi
ntocont
ainer
s
wi
thasmal
lopeni
ngwi
thoutspi
l
li
ng.Usedf
orf
il
tr
ati
onort
hedel
i
ver
yofl
i
qui
ds.
9. Di
ssect
ingpan-i
tisapanwher
eyoudi
ssector
gani
smssot
heor
gansorf
lui
d
wi
l
lnotspr
eadaway
.
10.Gl
asssl
ide-i
tisusedt
opl
aceaspeci
menonandputi
tunderami
croscopet
o
v
iew
11.Dr
opper–usedf
oraddi
ti
onofl
i
qui
ds,
dropbydr
op
12.Di
ssect
ingsci
ssor
s-i
tisusedf
orsl
i
cingaspeci
mendur
ingdi
ssect
ion
13.
For
ceps-i
tisusedf
orgr
aspi
ng,mani
pul
ati
ngorext
ract
ing,especi
all
ysuchan
i
nst
rumentusedbyasur
geon;
hol
dorpi
ckupsmal
lobj
ect
sthatar
etoosmal
lfor
y
ourhandsorf
inger
s;hol
dti
ssueoutoft
hewayort
opi
ckupast
ruct
ure.
For
cepsi
susedt
ohol
dorpi
ckt
hingst
hatar
etoosmal
lfory
ourhandsorf
inger
s.
14.Scal
pel-i
tisaf
ixedbl
adeusedi
nsur
ger
yanddi
ssect
ion(
isanext
remel
yshar
p
bl
adedi
nst
rumentt
hatcanneat
lyspl
i
topenski
nandcutt
hroughmuscl
eand
or
gans)
15.Beaker-i
sasi
mpl
econt
ainerf
orst
ir
ri
ng,mi
xi
ngandheat
ingl
i
qui
dscommonl
y
usedi
nmanyl
abor
ator
ies.I
tisal
sousedf
orhol
dingl
i
qui
dsi
nthel
abor
ator
y,a
l
i
qui
d-measur
ingcont
ainer
;usedwhent
hesol
uti
onmi
xedi
niti
sgoi
ngt
obe
pour
edi
ntosomet
hingel
se.Theyhav
eal
i
pont
hem f
orpour
ing.

16.
Flasks-hav
eanar
rowneckandar
eusedwhent
hesol
uti
onmayspl
ashoutofa
beakerorwhent
hecont
ainerofsol
uti
onneedst
obepl
uggedatsomepoi
nti
n
t
heexper
iment
.
17.Testt
uber
ack-i
tisusedt
ohol
dthet
estt
ubesot
hesol
uti
on/
subst
ancei
nit
won’
tspr
eadaway
18.Di
ssect
ingpi
n-i
tisusedt
ohol
dopent
hespeci
ment
haty
ouar
edi
ssect
ing
19.Met
erst
ick– i
tisast
ickt
hati
s100cent
imet
ersl
ong,r
oughl
y39i
nches.
Gener
all
yusedt
omeasur
edi
stance.
20.
Bunsen bur
ner– i
sheatsour
ce;acy
li
nderi
sat
tached t
o agasl
i
ne.i
tis
commonl
yusedi
nsci
ent
if
icl
abor
ator
ies;consi
sti
ngessent
ial
l
yofahol
l
owt
ube
whi
chi
sfi
tt
edv
ert
ical
l
yar
oundt
hef
lame&whi
chhasanopeni
ngatt
hebaset
o

4
 admi
tai
r.Whent
hegasl
i
nei
sopened,aspar
kigni
tesaf
lamei
ntheBunsen
bur
ner
,whi
chi
sthenusedt
oheatsol
uti
ons.
21.
Tri
ple-
beam bal
ance-i
tisusedt
ofi
ndt
hemassofanobj
ect
22.
Saf
etygoggl
es-i
tisusedaspr
otect
ionofey
efr
om dust
,fl
yi
ngdebr
is,
chemi
cal
spl
ashes,
etc.
23.
Pipet
te-Api
eceofl
abor
ator
ygl
asswar
e,shapedl
i
keat
hint
ubewi
thabul
gei
n
t
hemi
ddl
e,t
hatal
l
owsbet
teraccur
acywhenmeasur
ingcer
tai
nvol
umes(
hence
t
her
angeofsi
zes)e.
g.whenmaki
ngsol
uti
onsorsampl
esf
ort
it
rat
ion.Theyar
e
usedi
nal
abor
ator
ytot
ranspor
tand/
ormeasur
easpeci
fi
cvol
umeofl
i
qui
d.
24.
Saf
etymask–i
tisamaskusedwhendoi
ngexper
iment
,di
ssect
ionorsur
ger
yto
pr
otectusf
rom poi
sonousgasses,
vir
usesanddust
s.
25.
HotPl
ate-
Usedf
orheat
ingl
i
qui
ds

26.
Erl
enmey
erFl
asks-hav
eanar
row neckandar
eusedwhent
hesol
uti
onmay
spl
ashoutofabeakerorwhent
hecont
ainerofsol
uti
onneedst
obepl
uggedat
somepoi
nti
ntheexper
iment
;ar
eusedt
omeasur
e,mi
x,andst
orel
i
qui
ds.May
beusedt
ohol
dli
qui
ds,i
nst
eadofbeaker
s,whenasmal
l
eropeni
ngi
spr
efer
red.
Themostcommonsi
zesofer
lenmey
erf
laskspr
obabl
yar
e250mland500ml
.
Theycanbef
oundi
n50,
125,
250,
500,
1000ml
.
27.
Measur
ingcy
li
nder
s–
28.
Vor
texmi
xer-r
otat
est
hebot
tom ofat
uber
api
dly
;set
ti
ngupav
ort
exi
nthe
l
i
qui
dthatr
api
dlymi
xest
hecont
ent
.

29.
Tong-usedt
ohandl
ehotbeaker
s
30.
Testt
ubehol
der–spr
ingmet
alusedt
ohol
dtestt
ubesorgl
asses
31.
Dissect
ingpi
n-i
tisusedt
ohol
dopent
hespeci
ment
haty
ou'
redi
ssect
ing
32.
Vol
umet
ri
cFl
ask-Of
tenusedwhensol
uti
onsofspeci
fi
cconcent
rat
ionar
ebei
ng
made.
33.
Graduat
edCy
li
nder
-Usedt
omeasur
ethev
olumeofl
i
qui
ds
34.
DropperPi
pet
te-Usedt
otr
ansf
ersmal
lamount
sofl
i
qui
d/usedt
otr
ansf
er
measur
edsubst
ancesi
ntoanot
herv
essel
35.
GlassSt
ir
ri
ngRod-
Usedt
ost
irl
i
qui
ds

5
36.
Aut
ocl
ave–usedf
orst
eri
l
izat
ion.
37.
Cent
ri
fuge-i
samachi
net
hatspi
nst
estt
ubesatahi
ghspeed.Thi
scent
ri
fugal
f
orcepul
l
spar
ti
clest
othebot
tom oft
het
estt
ubes,
separ
ati
ngpar
tsofacol
l
oid.
I
tisusedt
osepar
atebl
oodcel
l
sfr
om t
hepl
asma.Theyar
eusedi
nsepar
ati
ng
anysubst
ances.
38.
Ult
rav
iol
et/
visi
ble(
UV/
VIS)spect
rophot
omet
er-measur
esabsor
banceofl
i
ght
i
nthe
ul
tr
avi
oletr
egi
onoft
hespect
rum (
about100-
700nm)
39.Mi
cropi
pet
tes–dev
icesr
out
inel
yusedt
omeasur
eandt
ransf
erl
i
qui
dsby
dr
awi
ngt
hel
i
qui
dont
op
40.El
ect
roni
cbal
ances-hav
erepl
acedmostmechani
cal
bal
ancesduet
othei
r
gr
eat
eraccur
acyandeaseofoper
ati
on
41.
Col
umn chr
omat
ogr
aphy – used f
or separ
ati
on of di
ff
erent chemi
cal
compounds.
42.
Incubat
or-i
sadev
iceusedt
ogr
owandmai
ntai
nmi
crobi
ologi
calcul
tur
esorcel
l
cul
tur
es?The i
ncubat
ormai
ntai
ns opt
imalt
emper
atur
e,humi
dit
yand ot
her
condi
ti
ons such as t
he car
bon di
oxi
de (
CO2)and oxy
gen cont
entoft
he
at
mospher
einsi
de.
43.
Incubat
ingshaker–i
ncor
por
ateO2 &ev
enl
ydi
str
ibut
enut
ri
ent
sthr
oughoutt
he
cul
tur
emedi
a
44.
Wat
erdi
sti
ll
ati
onuni
t-usedf
orwat
erpur
if
icat
ionanddi
sti
l
lat
ion
45.
Biosafet
ycabinet-i
sanenclosed,venti
latedlaborat
oryworkspacef
orsaf
ely
workingwit
hmat eri
alscont
aminatedwith( orpotent
ial
lycont
aminat
edwit
h)
pathogensrequi
ri
ngadef i
nedbiosafet
yl evel

46.
pHmet
er-measur heamountofH+i
est onsi
nasol
uti
onandi
sameasur
eof
howaci
dicasol
uti
oni
s/measur
esaci
dit
yofsol
uti
ons
47.
Ref
ri
ger
ator-usedf
ort
hest
orageoft
hest
ocksol
uti
ons,
chemi
cal
s,ki
tsand
PCRpr
oduct
sthatshoul
dbemai
ntai
nedatcer
tai
ntemper
atur
es
48.
Bur
ett
e-agr
aduat
edgl
asst
ube,
commonl
yhav
ingast
opcock(
val
vef
or
cont
rol
l
ingf
low)att
hebot
tom,
usedf
oraccur
atel
ymeasur
ingormeasur
ingout
smal
lquant
it
iesofl
i
qui
d.

6
 49.
Wat
erBat
h-i
sadev
icet
hatmai
ntai
nswat
erataconst
antt
emper
atur
e(usedf
or
r
egul
ati
ngt
het
emper
atur
eofsubst
ancessubj
ect
edt
oheat
).I
tisusedi
nthe
mi
crobi
ologi
cal
labor
ator
yfori
ncubat
ions.
50.
Aquar
ium -acont
ainer(
asagl
asst
ank)oranar
ti
fi
cial
pondi
nwhi
chl
i
ving
aquat
icani
mal
sorpl
ant
sar
ekept
51.
Ther
mal
cycl
er(
PCRmachi
ne)
.Thi
sdev
icei
susedf
ort
heampl
i
ficat
ionofa
speci
fi
cregi
onofanyDNAsampl
ewi
thpol
ymer
asechai
nreact
ioni
nat
estt
ube.
I
tisal
sousedf
ordet
ect
ionandconst
it
uti
onofgenet
ical
l
ymodi
fi
edor
gani
sms,
aswel
lot
hergenet
icanal
yses.
52.
Lami
narf
lowhood:
prov
idesanasept
icwor
kar
eawhi
l
eal
l
owi
ngt
he
cont
ainmentofi
nfect
iousspl
ashesoraer
osol
sgener
atedbymany
mi
crobi
ologi
cal
procedur
es.

Quest
ions
1.Whatar
ethepr
operpr
act
icesf
orwor
kingsaf
etyi
nal
abor
ator
y?
2.Whymi
ghteat
ingordr
inki
ngi
nthel
abor
ator
ybedanger
ous?
3.Lookar
oundt
her
oom.Whatsaf
etyequi
pmentdoy
our
ecogni
ze?
4.Whatki
ndsofmeasur
ement
smi
ghty
ouneedt
omakei
nthel
abor
ator
y?Whatki
nds
ofequi
pmentwoul
dyouneedf
ort
heset
asks?

Bur
ett
e Cent
ri
fuge

7
 Pet
ridi
sh Magni
fyi
nggl
ass(
handl
ens)

Cov
ersl
i
p Ther
momet
er

\
Testt
ubes Vor
texmi
xer

8
Mi
crocent
ri
fugeHol
dsEppendor
f,ormi
crocent
ri
fuge Funnel

Ani
mal
dissect
ionondi
ssect
ingpan Di
ssect
ingsci
ssor
s

For
ceps(
tweezer
s) Scal
pel

9
 Testt
uber
ack Beaker
s

Bunsenbur
ner Pi
pet
te

Tr
ipl
ebeanbal
ance Hotpl
ate

10
 Er
lenmey
erf
lasks Vol
umet
ri
cFl
ask

Gr
aduat
edcy
li
nder
s Dr
opperpi
pet
te

Glassstr
ingrod Ul
tr
avi
olet
/vi
sibl
e(UV/
VIS)
spect
rophotometer

11
 Aut
ocl
ave El
ect
roni
cbal
ance

Mi
cropi
pet
tes Lami
narf
lowhood

12
 I
ncubat
or I
ncubat
ingshaker

Wat
erdi
sti
l
lat
ionuni
t Ther
mal
cycl
er(
PCRmachi
neorDNAampl
i
fier
)

Wat
erbat
h Tong

13
Testt
ubehol
der Di
ssect
ingpi
ns

Fi
gur
e1.
1:Somebasi
cequi
pmentusedi
nbi
ologyl
ab

LABORATORYSTUDYNUMBER2

MI
CROSCOPES

2.
1Int
roduct
ion
Mi
croscopesar
eusef
ulf
orv
iewi
ngobj
ect
sthatar
etoosmal
ltoseecl
ear
lywi
thout
magni
fi
cat
ion.Mi
croscopesar
ethemosti
mpor
tantsci
ent
if
ici
nst
rument
susedmai
nly
bybi
ologi
sts.Ther
ear
etwodi
ff
erentt
ypesofmi
croscopes:l
ightmi
croscopesand
el
ect
ronmi
croscopes.

Li
ghtmi
croscopes
Li
ghtmi
croscopeshav
egl
assl
enseswhi
chmagni
fyobj
ect
sandusel
i
ghtt
oil
l
umi
nat
e

14
t
heobj
ect
sbei
ngexami
ned.Thesmal
l
estsi
zey
oucanseewi
thy
ournakedey
eis0.
2
mm,whi
chi
sequalt
o200mi
cromet
ers.Thi
ssi
zei
sequi
val
entt
ooner
idgeony
our
f
inger
pri
nt.Li
ghtmi
croscopesmagni
fycel
l
supt
o1,
000t
imes.Usi
ngt
heshor
testr
ayof
l
i
ght
,whi
chal
l
owst
hehi
ghestr
esol
uti
on,
li
ghtmi
croscopescanv
iewt
hingsassmal
las
0.
2mi
cromet
ersi
nwi
dth,
i.
e.,
0.0002mm.

Youwi
l
lbeusi
ngt
wodi
ff
erentki
ndsofl
i
ghtmi
croscopesi
nthi
slab,
thecompound
mi
croscopeandt
hedi
ssect
ingmi
croscope.

CompoundMi
croscopes
Compoundmi
croscopesar
eusedt
oexami
neobj
ect
sint
wodi
mensi
ons.Theyuset
he
magni
fyi
ngpower
soft
woconv
exl
ensest
opr
oduceamagni
fi
edi
mageofav
erysmal
l
obj
ect
.Ver
ysmal
lor
gani
smsorcr
oss-
sect
ionsofor
gani
smsar
epl
acedoncl
eargl
ass
sl
i
des;t
hese obj
ect
s ar
evi
ewed as l
i
ghtpasses t
hrough t
hem.The compound
mi
croscopei
sthemostcommonl
yusedmi
croscopeandi
tispower
fulenoughsot
hat
t
heusercanv
iew cel
l
s,ev
ent
hel
i
vingones.Themagni
fi
cat
ionoft
hismi
croscopei
s
hi
ghbutt
her
esol
uti
oni
squi
tel
ow.

Di
ssect
ingMi
croscopes
Di
ssect
ingmi
croscopesar
eusedt
oobser
vemat
eri
alt
hati
sei
thert
oot
hickort
ool
arge
t
obev
iewedwi
tht
hecompoundl
i
ghtmi
croscope.Wi
tht
hesemi
croscopes,
youseet
he
sur
faceoft
hingst
hatr
efl
ectt
hel
i
ght
.Whi
l
ethemagni
fi
cat
ionanddept
hoff
iel
dar
e
smal
l
eri
nthedi
ssect
ingscope,
thef
iel
dofv
iewi
smuchl
arger
.Asi
tsnamei
mpl
i
es,
the
di
ssect
ingscopei
sof
tenusedt
ost
udyst
ruct
ures(
li
keant
ennaeandl
egsofi
nsect
s)
l
ookatpar
tsofor
gani
sms(
li
kei
nsect
s)asy
oudi
ssectt
hem,si
ncei
tal
l
owsf
or
mani
pul
ati
onofmat
eri
al.Mostoft
hepar
tsoft
hedi
ssect
ingmi
croscopear
ethesame
ast
hecompoundmi
croscope
El
ect
ronmi
croscopes
Forobj
ect
ssmal
l
ert
han0.
2 mi
cromet
ers,anel
ect
ronmi
croscopemustbeused.
El
ect
ronmi
croscopesal
l
ow y
out
oseeobj
ect
sthatar
eassmal
las0.
2nanomet
ers
(
nm)
,whi
chi
sequalt
o0.
000000002mm.I
ncompar
isont
oal
i
ghtmi
croscopebei
ng
abl
etomagni
fy1,
000t
imes,
elect
ronmi
croscopescanmagni
fyobj
ect
s200,
000t
imes.

15
El
ect
ronmi
croscopesusebeamsofel
ect
ronst
oexami
nei
ncr
edi
blysmal
lobj
ect
s(l
i
ke
t
hecomponent
sofani
ndi
vi
dualcel
l
)thathav
ebeenspeci
all
ypr
epar
ed.I
nthese
mi
croscopesabeam ofel
ect
rons(
inpl
aceofl
i
ght
)andci
rcul
armagnet
s(i
npl
aceof
gl
assl
enses)per
mitt
her
esol
uti
onofst
ruct
uresi
nmuchf
inerdet
ailt
hani
nanopt
ical
mi
croscope.

Ther
ear
etwot
ypesofel
ect
ronmi
croscope.Thesear
escanni
ngel
ect
ronmi
croscope
(
SEM)andt
ransmi
ssi
onel
ect
ronmi
croscope(
TEM)
.

Scanni
ngEl
ect
ronMi
croscope(
SEM)
:abeam ofel
ect
ronsscanst
hesur
faceofan
opaqueobj
ectandpr
oducesani
mageoft
hatsur
face.Thi
sisanexpensi
vemi
croscope
whi
chhashi
ghmagni
fi
cat
ionandr
esol
uti
on.Thei
magesar
evi
ewedonacat
hodet
ube,
ori
npi
ctur
est
akenwi
tht
hemi
croscope.Manyoft
hephot
ogr
aphsofcel
lst
ruct
ure
usedi
nyourt
extwer
etakenwi
thanel
ect
ronmi
croscope.

Tr
ansmi
ssi
onEl
ect
ronMi
croscope(
TEM)
:anel
ect
ronbeam passest
hrought
he
speci
menandpr
ovi
dest
heuserwi
that
wodi
mensi
onalv
iew andhi
ghmagni
fi
cat
ion
andr
esol
uti
on.

A B

16
 C D
Fi
gure2.1:
Di ff
erenttypesofmicr
oscopes.A.Compoundl
ightmicroscope;B.
Di
ssecti
nglightmi cr
oscope;C.Transmissi
onelect
ronmi
croscope;D.Scanni
ng
el
ect
ronmi croscope

Obj
ect
ives:
Att
heendoft
hissect
iont
hest
udent
swi
l
lbeabl
eto:
 descr
ibet
hepar
tsandf
unct
ionsoft
hecompoundmi
croscope
 uset
hemi
croscopebyf
ocusi
ngwi
tht
hedi
ff
erentobj
ect
ives

2.
2Thepar
tsandf
unct
ionsofacompoundmi
croscope
A compoundmi
croscopei
ssocal
l
edbecausei
tusesacombi
nat
ionofl
ensest
o
magni
fyanysor
tofi
mages.Basi
cal
l
y,t
her
ear
etwo component
s-t
hest
ruct
ural
component
sandt
heopt
ical
component
s.

St
ruct
uralcomponent
s
Ast
henamesuggest
s,t
hesear
ethecomponent
sthatmakeupt
heconf
igur
ati
onofa
compoundmi
croscope.Ther
ear
ethr
eebasi
cst
ruct
uralcomponent
sofacompound

17
mi
croscope.

1.HeadorBodyt
ube
Bodyt
ubeorey
epi
ecet
ubei
sthepar
tthatconnect
stheocul
arst
otheobj
ect
ives.I
nthi
s,
ocul
arsar
epl
acedatt
heupperpor
ti
onabov
etheobj
ect
ives.

Base-Basei
sthebot
tom par
t,whi
chsuppor
tst
hemi
croscope.I
ll
umi
nat
ori
slocat
edi
n
t
hebase.
Ar
m -Ar
m suppor
tst
heheadcomponentandconnect
sitt
othebase.
I
tisal
way
srecommendedt
ohandl
ebyt
hear
m andsuppor
tthebasewhi
l
ecar
ryi
nga
compoundmi
croscope.

2.Opt
ical
Component
s
Wor
kingofami
croscopeent
ir
elydependsont
heopt
icalcomponent
s.Theopt
ical
component
sar
efi
xedt
other
igi
dar
m oft
hemi
croscope.
Themi
croscopehast
wo magni
fyi
ng l
enses:t
heey
epi
eceorocul
arl
ensand t
he
obj
ect
ivel
ensesonat
urr
etwhi
chr
evol
vesabov
ethest
age.
Ey
epi
eces(
Ocul
ars)
Ey
epi
eceorocul
arsar
esecondar
ylensesl
ocat
edatt
het
opoft
heBODYTUBE,
neart
he
ey
e.I
tist
hrought
hisl
enst
hatwev
iew anobj
ecti
nami
croscope.Themagni
fi
cat
ion
powerofey
epi
ecel
ensesi
susual
l
y10X.

Obj
ect
ivel
enses
Obj
ect
ivesar
ethepr
imar
ylensesofacompoundmi
croscope.Theobj
ect
ivel
enses
(
ther
ear
efour
:4X,10X,40Xand100X)ar
escr
ewedi
ntoar
evol
vi
ngnosepi
ece.By
changi
ngt
heobj
ect
ivest
heef
fect
ivepowerofmagni
fi
cat
ioni
schanged.Thet
otal
magni
fi
cat
ionobser
vedi
sthepr
oductoft
hepowerofmagni
fi
cat
ionoft
heey
epi
ece
andt
heobj
ect
ive.Onl
ythe100Xobj
ect
ivei
susedi
mmer
sedi
nadr
opofspeci
aloi
l
(
bet
weent
hel
ensandt
hesl
i
de;
allot
her
sar
edesi
gnedt
obeusedwi
thai
rbet
weent
he
obj
ectandl
enssur
face.Thepowerofmagni
fi
cat
ioni
scl
ear
lyi
ndi
cat
edoneachl
ens

18
al
ongwi
tht
henumer
ical
aper
tur
eofeachl
ens.Dependi
ngupont
hei
rdesi
gnandqual
i
ty,
di
ff
erentobj
ect
ives hav
e di
ff
erentr
esol
vi
ng di
stances.The l
att
eri
sthe smal
l
est
di
stancebet
weent
wopoi
ntst
hatal
l
owsbot
hpoi
ntst
obev
iewedassepar
ate.Thi
s
r
esol
vi
ngdi
stancei
sdependentupont
hewav
elengt
hofl
ightusedaswel
last
he
const
ruct
ionoft
hel
ens.

Exami
net
heobj
ect
ivesf
rom t
hesi
deandnot
ethemagni
fi
cat
ion.

A.Thel
owpowerobj
ect
ive-t
heshor
testonewi
tht
hel
argestl
ensopeni
ng)
,usual
l
y
hasamagni
fi
cat
ionoff
ourt
imesandi
slabel
edas4X.
B.Themi
ddl
epowerobj
ect
ive–i
smedi
um i
nlengt
handof
tenhasamagni
fi
cat
ion
of10X.
C.Thehi
ghpowerobj
ect
ive-i
slongerandhasamagni
fi
cat
ionof40X.
D.Theoi
limmer
sionobj
ect
ivehasamagni
fi
cat
ionof100X.

3.Nosepi
ece-Nosepi
ecesuppor
tst
heobj
ect
ives.Theobj
ect
ivesar
emount
edona
r
otat
ingt
urr
et sot
hatanyoft
heobj
ect
ivescanbeconv
eni
ent
lyusedf
orv
iewi
ng.
4.Adj
ust
mentKnobs-Themi
croscopei
spr
ovi
dedwi
tht
wof
ocusi
ngmechani
sms,
the
cour
seadj
ust
mentandt
hef
ineadj
ust
ment
.Thesear
eusedf
orf
ocusi
ng,
soast
ogeta
cl
earef
fect
.Thef
ocusi
ngknobsmov
ethel
ensassembl
yupanddownt
obr
ingt
he
obj
ecti
nfocus.Thecoar
seadj
ust
mentshoul
donl
ybeusedwi
tht
heshor
test
,low
powerobj
ect
ivel
ens.Thef
ineadj
ust
ment(
smal
l
erknob)br
ingst
heobj
ecti
ntocr
it
ical
f
ocus.Not
icet
hatal
lobj
ect
sar
epr
oject
edupsi
dedowni
nthemi
croscopef
iel
d.I
ttakes
al
i
ttl
epr
act
icei
nusi
ngt
hemechani
cal
staget
omov
ethesl
i
dewher
eyouwanti
t.

Theadj
ust
mentknobsofy
ourmi
croscopear
eusedasf
ocusi
ngdev
icesbecauset
hey
canr
aiseorl
owert
hebodyt
ubeasnecessar
y.I
nsomemi
croscopes(
e.g.
,Rei
cher
t
Neov
ar)i
tist
hest
agebutnott
hebodyt
ubet
hati
slower
edorr
aisedi
nfocusi
ng.

Not
ethatt
heposi
ti
onsoft
headj
ust
mentknobsv
arywi
tht
hemanuf
act
urer
s.The
BauschandLombmi
croscopey
oupossesshasi
tscour
seadj
ust
mentknobatt
het
op
oft
her
am andi
tsf
ineadj
ust
mentknobl
ocat
edatt
hepi
votofi
ncl
i
nat
ionj
oint
.

19
I
nsomet
ypesofmi
croscopes(
e.g.
,Rei
cher
tNeov
ar)t
hecoar
seadj
ust
mentandf
ine
adj
ust
mentar
einonepi
ece.Thei
nnerl
argerpar
tist
hecoar
seandout
ersmal
l
erpar
tis
t
hef
ineadj
ust
ment
.Inst
il
lanot
hert
ypeofmi
croscope(
e.g.
,Rei
cher
tMonopan)t
he
coar
seandt
hef
ineadj
ust
ment
sar
ecombi
nedi
ntoone.I
nthi
scaset
hedi
rect
ionoft
he
mov
ement(
ther
otat
oryorbackandf
ort
hmov
ement
)oft
heknobdet
ermi
nest
ypeof
adj
ust
ment
.

St
age
St
agei
sthepl
atf
orm,
wher
ethesl
i
det
obev
iewedi
spl
aced.Ther
eisanopeni
ngi
nthe
cent
reoft
hest
age(
stageopeni
ng)t
hroughwhi
chl
i
ghtpasses.Ont
hest
agest
her
ear
e
t
wo met
alcl
ips t
hatser
vet
o hol
dthe sl
i
de secur
elyi
n pl
ace.I
nst
ead some
mi
croscopesar
eequi
ppedwi
thmechani
caldev
icet
hathol
dst
hesl
i
dei
npl
aceandal
so
t
omov
ethesl
i
desi
deway
saswel
lasbackandf
ort
h.Forhi
ghermagni
fi
cat
ionst
udi
es,
mechani
cal
stagei
susedf
orf
inemov
ementoft
hesl
i
de.

Condenser
Mi
croscopescont
ainel
ement
sdesi
gnedt
opr
ojectpar
all
elbeamsofl
i
ghtt
hrought
he
speci
menandi
ntot
heobj
ect
ive.Thesei
ncl
udet
hepr
oject
ionl
enswhi
chf
ocusesl
i
ght
ont
othecondenserl
ens.Thecondenserl
ensf
ocusesl
i
ghtont
otheobj
ect
.

The Condenseri
sfound i
mmedi
atel
ybeneat
hthe st
age.Thi
s componentoft
he
mi
croscopecausest
hel
i
ghtr
ayst
oconv
ergeatt
hest
ageopeni
ng.Ati
tsl
owerendt
he
condenseri
sequi
ppedwi
thani
ri
sdi
aphr
agm whi
chcanbeopenedandcl
osedas
necessar
ytocont
rolt
heamountofl
i
ghtent
eri
ngt
hebodyt
ube.Thecondensermaybe
f
ixed ori
tmaybe r
aised and l
ower
ed bya sub-
stage adj
ust
ment
.Lower
ing t
he
condenserdecr
easest
heamountofl
i
ghtent
eri
ngt
hebodyt
ubeandul
ti
mat
elyr
eachi
ng
y
ourey
e.

Aper
tur
e-Thehol
eint
hest
aget
hroughwhi
chl
i
ghtr
eachest
heobj
ect
.
I
ll
umi
nat
or-I
ll
umi
nat
ori
sthesour
ceofl
i
ghti
nami
croscope.I
tispl
acedi
nthebase
component
.

20
 Fi
gur
e2.
2:Par
tsofacompoundmi
croscope

Mi
croscopeHandl
ing

1. Al
way
scar
ryt
hemi
croscopei
nast
rai
ghtupr
ightposi
ti
onwi
thonehandar
oundt
he
ar
m andt
heot
herhandundert
hebase.Theey
epi
ecesar
enotat
tachedandwi
l
lfal
l
outi
fthemi
croscopei
scar
ri
edatanangl
eorupsi
dedown.
2.Keept
hemi
croscopecl
ean.Whenany
thi
ngi
sspi
l
ledorot
her
wiseget
sont
he
mi
croscope,
cleani
tupi
mmedi
atel
y.
3.Whenusi
ngt
hemi
croscopest
artwi
tht
hel
owpowerl
ensandwor
kupt
othedesi
red
magni
fi
cat
ion.Thesemi
croscopesar
eparf
ocal
,whi
ch meanst
hatal
lpower
s
shoul
dbei
nfocuswhent
het
urr
eti
srot
ated.
4.Nev
ermov
ethest
ageupwar
dswi
tht
hecoar
seadj
ust
mentwhi
l
evi
ewi
ngt
hrough
t
heey
epi
eces.Gett
hel
enscl
oset
othesl
i
dewhi
l
evi
ewi
ngf
rom t
hesi
det
omake

21
 sur
ethatt
heynev
ert
ouch.Then mov
ethe st
age downwar
d wi
tht
he coar
se
adj
ust
mentwhi
l
evi
ewi
ng t
hrough t
he l
ens.Thi
swi
l
lpr
eventt
he possi
bil
i
tyof
r
ammi
ngt
hel
ensi
ntot
hesl
i
de,
ther
ebyr
uini
ngasl
i
dey
ouhav
ejustmadeand,
qui
te
possi
bly
,damagi
ngt
hel
ens.
5.Donott
oucht
hel
enses
Cl
eant
hel
enseswi
thl
enspaperonl
y.Donotcl
eant
hel
enseswi
thhandker
chi
efs,
f
aci
alt
issues,
papert
owel
s,et
c.-t
heywi
l
lscr
atcht
hel
enses.I
fyourl
ensesar
ever
y
di
rt
y,obt
ainsomel
enscl
eani
ngsol
ventf
rom t
hei
nst
ruct
or.
6.Moi
st,
li
vi
ngorpr
eser
vedmat
eri
alsmustbeobser
vedt
hroughacov
ersl
i
p.Thi
s
pr
otect
sthel
ensaswel
last
endst
omaket
heobj
ectunderv
iewopt
ical
l
yfl
at.Be
sur
etomai
ntai
nasaf
edi
stancebet
weent
hecov
ersl
i
pandt
heobj
ect
ivel
enses.

7.Remov
ethedustcov
erandst
orei
tpr
oper
ly.Pl
ugi
nthescope.Donott
urni
tonunt
il
t
oldt
odoso.
8.Teacher
sandst
udent
sshoul
dbesur
etokeepal
lGl
assandShar
pinst
rumentSaf
ety
Rul
est
hatar
egi
venbyt
het
eacherandi
nal
lgener
all
abor
ator
yor
ator
yexper
iences
whenpr
epar
ingmi
croscopesl
i
des.
9. Al
way
scl
eansl
idesandmi
croscopewhenf
ini
shed.Tur
nof
fli
ght
.
Cov
ert
hemi
croscopewi
thdustcov
erandr
etur
nyourmi
croscopet
othecabi
net
wi
thl
i
ghtcor
dwr
appedar
oundi
tsbaseandwi
tht
hel
owestpowerobj
ect
ivel
ens
i
nposi
ti
on,
ifr
equest
edbyt
eacher
.

2.
3Pr
eli
minar
yuseoft
hemi
croscope

Set
ti
ng-
upmi
croscopef
oruse

I
nor
dert
ost
udyy
ourmount
eddotoranyot
herobj
ectundert
hemi
croscopef
ir
sty
ou
hav
etoset
-upy
ourmi
croscopef
oruse.
I
fyoufol
l
owthefol
l
owingsteps,
theywi
llenabl
eyousetupyourmicr
oscopeforuse.
1.Pl
acethemicr
oscopeneartheedge,notontheedge,ofy
ourlabor
atorybench
wi
thi
tsar
mfaci
ngy
ou.
2.Ar
rangey
ourl
abor
ator
yst
oolsucht
hati
tal
l
owsy
ouaneasyl
ookt
hrought
he
ocul
arofy
ourmi
croscopewi
thouti
ncl
i
ningi
t.

22
 3.Maket
hedi
aphr
agm wi
deopen
4.I
fthecondenseri
sinal
ower
edposi
ti
on,
rai
sei
tasneart
hest
ageasi
twi
l
lgo
5.Tur
nony
ourmi
croscopet
hroughandwhi
l
elooki
ngi
ntot
hemi
croscopewi
thout
t
heocul
aradj
ust
stheconcav
esur
faceoft
hemi
rr
orunt
ilt
hef
iel
dofv
isi
on(
the
ci
rcul
arar
eay
ouseei
nthemi
croscope)i
suni
for
mlyi
l
lumi
nat
ed.Not
ethatonce
y
ouhav
eadj
ust
edt
hel
i
ghty
oushoul
dnotmov
ethemi
croscopeort
hel
amp.
Ot
her
wisey
oumayneedt
oreadj
ustt
hel
i
ght
.

I
.Pr
act
icei
nmount
ingandf
ocusi
ng

Mat
eri
alsandchemi
cal
s
Pen/
penci
l
,cov
ersl
i
ps,
glasssl
i
des,
micr
oscope,
Oni
on,
met
hyl
enebl
uest
ain,
wat
er

1.Dr
ymountsl
ide:

1.Pl
aceasampl
eint
hecent
eroft
hesl
i
de
2.Car
eful
l
ylowert
hecov
er-
sli
pont
othesl
i
de-donotdr
opt
hecov
er-
sli
p
3.Secur
eyoursl
i
det
othest
age(
wit
hthespr
ingcl
i
ps)
4.Focusont
hesampl
e

2.Wetmountsl
ide
Wetmount
sar
ecommonl
yused,
forexampl
e,t
ovi
ewmi
croscopi
cor
gani
smst
hatgr
ow
i
npondwat
erorot
herl
i
qui
dmedi
a,especi
all
ywhenst
udy
ingt
hei
rmov
ement
,si
zeof
cel
l
sandbehav
ior
Thespeci
meni
spl
acedi
nadr
opofwat
erorot
herl
i
qui
dhel
dbet
weent
hesl
i
deandt
he
cov
ersl
i
p

1.Pl
aceasampl
eint
hecent
eroft
hesl
i
de
2.Addadr
opofwat
eront
hesampl
e
3.Addasmal
ldr
opofmet
hyl
enebl
uest
aint
othepr
epar
ati
onandgent
lyagi
tat
e
4.Hol
dthecov
ersl
i
psot
hati
tsedget
ouchesonesi
deoft
hef
lui
d
5.Car
eful
l
ylowert
hecov
er-
sli
pont
othepr
epar
ati
on-donotdr
opt
hecov
er-
sli
p
6.Secur
eyoursl
i
det
othest
age(
wit
hthespr
ingcl
i
ps)

23
 7.Focusont
hesampl
ethr
ought
heey
epi
eces

Fi
gur
e2.
3:Techni
queofappl
yi
ngcov
ersl
iponspeci
menont
hegl
asssl
ide

A.Focusi
ngwi
thl
owpower

1.I
fthel
owpowerobj
ect
ivei
snoti
nposi
ti
on(
i.
e.i
fiti
snotr
ightabov
ethest
age
openi
ng)hol
danyt
woobj
ect
ivesbet
weeny
ourt
humbandt
hef
oref
ingerandt
urn
t
hepi
eceunt
ilt
heobj
ect
ivecl
i
cksi
ntoposi
ti
ons.
2.Looki
ngf
rom t
hesi
de,l
owert
hebodyt
ubewi
tht
hecoar
seadj
ust
mentt
owi
thi
n
5mm oft
hesl
i
de.Thebodyt
ubeofsomet
ypesoft
hemi
croscopescannotbe
l
ower
edt
otheext
entt
hel
ow powerobj
ect
ivet
ouchest
hesl
i
de.Thus,i
fyou
cannotl
oweri
tfur
ther
,donotappl
yfor
ce.

 I
nsomet
ypesofmi
croscopei
tist
hest
aget
hati
srai
sedupt
owar
dst
he
obj
ect
ivei
nst
ead.
3.Looki
ngi
ntot
hemi
croscopeabout2cm awayf
rom t
heey
epi
ece,
slowl
yrai
set
he
bodyt
ubewi
tht
hecoar
seadj
ust
mentunt
ilt
hei
magecomesi
ntov
iew.Donot
pl
anty
ourey
eri
ghtont
heocul
arf
ori
fyouseeany
thi
ngy
ouwi
l
lonl
yseea
r
efl
ect
ionofy
ourey
eandey
elashes.Keepbot
hey
esopenwhi
l
elooki
ngi
ntot
he
mi
croscope.
 Spect
acl
esusedt
ocor
rectnearorf
ar-
sight
ednessneednotbewor
nwhenusi
ng
t
hemi
croscope.Onl
ythosewhoar
ever
ynear
-si
ght
edmayneedspect
acl
est
o
av
oidheadaches.
4.I
fthei
magev
iewedi
sbl
urr
ed,f
ocusi
twi
tht
hef
ineadj
ust
mentunt
ili
tbecomes
cl
ear
.

5.I
fthei
magei
sof
fthecent
reoft
hef
iel
dofv
isi
on,
mov
ethesl
i
dev
erygent
ly,
whi
l
e

24
 l
ooki
ngi
ntot
hemi
croscope,
unt
ilt
hei
magei
sint
hecent
reoft
hef
iel
d.
6.Adj
ust(
incr
easeordecr
ease)t
hedi
aphr
agm asnecessar
y

B.Focusi
ngwi
thmi
ddl
epower
I
nor
dert
ofocust
heobj
ect
ives(
dot
)undert
hemedi
um powergot
ost
ep5wi
thout
changi
ngt
hef
ocusundert
hel
owpower
.
7.I
fyouhav
efol
l
owedt
heabov
est
eps,
yourobj
ecti
sinf
ocusundert
hel
owpower
.
Now,hol
dinganyt
wooft
heobj
ect
ives.Bet
weent
het
humbandt
hef
oref
inger
r
otat
ethenose-
pieceunt
ilt
hemi
ddl
epowerobj
ect
ivecl
i
cks(
i.
e.y
ouheara
cl
i
cki
ngsound)i
ntoposi
ti
on.
8.Focuswi
tht
hef
ineadj
ust
mentunt
ily
ougetacl
eari
mage.
9.I
ncr
easet
hedi
aphr
agm openi
ngi
fmor
eli
ghti
sneeded.

C.Focusi
ngwi
tht
hehi
gh-
power
Nowy
oucandi
rect
ly(
i.
e.wi
thoutchangi
ngt
hef
ocusi
ngundert
hemedi
um power
),go
t
ost
ep8i
nor
dert
ofocust
heobj
ect
iveundert
hehi
ghpower
.
10.
Yourobj
ecti
sal
readyi
nfocusundert
hemi
ddl
epower
.Taki
nghol
dofanyt
wo
obj
ect
ives,
rot
atet
henosepi
eceunt
ilt
hehi
ghpowerobj
ect
ivecl
i
cksi
ntoposi
ti
on.
Car
eshoul
dbet
akensot
hatt
heobj
ect
ivesdonothi
tagai
nstt
hecl
i
ps.
11.
Focuswi
tht
hef
ineadj
ust
mentunt
ily
ougetacl
eari
mage.Nev
eruset
hecoar
se
adj
ust
mentwhenf
ocusi
ngwi
tht
hehi
ghpowerobj
ect
ive.
12.
Ifmor
eil
l
umi
nat
ioni
sneededt
heopeni
ngoft
hedi
aphr
agm maybei
ncr
eased.
Youcanpr
act
icef
ocusi
ng(
steps1-
12abov
e)unt
ily
oudoi
tper
fect
ly.

D.Somei
mpor
tantpoi
ntsofcaut
ioni
nfocusi
ng
1.Nev
erf
ocusdownwar
dswi
thoutl
ooki
ngatt
heobj
ect
ivesf
rom t
hesi
dei
fyour
mi
croscopei
sthet
ypet
hati
sfocusedbyl
ower
ingorr
aisi
ngt
hebodyt
ube.

2.Nev
erf
ocusupwar
dswi
thoutl
ooki
ngatt
heobj
ect
ivesf
rom t
hesi
dei
fyour
mi
croscopei
sthet
ypet
hati
sfocusedbyl
ower
ingorr
isi
ngoft
hest
age.
3.Nev
erf
ocust
hehi
ghpowerobj
ect
iveort
heoi
limmer
sionobj
ect
ive.I
fyour
mi
croscopehasone,wi
thcoar
seadj
ust
ment
.Al
way
suset
hef
ineadj
ust
ment
.

25
 Ot
her
wise,
youmaybr
eakt
hesl
i
deandpossi
blydamaget
hemi
croscopei
tsel
f.
4.Donott
ryt
ofocusanobj
ect
ivedi
rect
lywi
tht
hehi
ghpowerobj
ect
iveorwi
tht
he
oi
limmer
sionobj
ect
ives.Fi
rstl
ocat
eorf
ocust
heobj
ectwi
tht
hel
ow power
,
t
henshi
ftt
othemedi
um powerandt
hent
othehi
ghpower
.Al
ter
nat
ivel
y,y
ou
canf
ocusi
twi
thl
ow powerandt
henhi
ghpowerwi
thoutf
ocusi
ngi
twi
tht
he
mi
ddl
epower
.
5.Youshoul
dnot
ewhi
chwayt
otur
ntheadj
ust
mentknobt
olowerorr
aiset
he
bodyt
ubeort
olowerorr
aiset
hest
agedependi
ngupont
het
ypeofy
our
mi
croscope.

26
2.
4.Ty
peofi
magef
ormed,
resol
uti
onandmagni
fi
cat
ion

A.Ther
esol
vi
ngpowerofmi
croscope

Resol
uti
onmeanst
heabi
l
ityt
osepar
atef
inedet
ail
s.Forexampl
e,i
fthedi
stance
bet
weent
wodot
sisl
esst
han0.
1mm,
thet
wodot
swoul
dappearasasi
ngl
edott
othe
unai
dedhumaney
e.Ther
esol
vi
ngpoweroft
hehumaney
eisabout0.
1mm.

Themi
croscopemagni
fi
esandr
esol
ves.Ther
esol
vi
ngpowerofami
croscopedepends
upont
heki
ndofi
ll
umi
nat
ionusedbecauset
her
esol
vi
ngpoweri
s½xt
hewav
elengt
h
oft
hei
l
lumi
nat
ionused.Ther
efor
e,ev
enwi
tht
hemostper
fect
lygr
oundl
ensesandwi
th
whi
tel
i
ghthav
inganav
eragewav
elengt
hof5500Angst
roms(
Å),t
heobj
ect
ivecannot
r
esol
veany
thi
ngl
esst
han2750Å(
i.
e.½*
5500or0.
275mi
crons)
.Si
ncemanypar
tsof
t
hecel
l
shav
edi
mensi
onsl
esst
han0.
275mi
crons(
or275mi
l
li
micr
ons)t
hei
rpr
esence
r
emai
nedundet
ect
edunt
ilameansofgr
eat
err
esol
uti
onwasf
ound.

Theel
ect
ronmi
croscopeusesbeam ofel
ect
ronsi
nst
eadofl
i
ght
.Whenel
ect
ronsar
e
pr
opel
l
edbyachar
geofabout50,
000v
olt
stheyhav
eawav
elengt
hofabout0.
05Å.
Ther
efor
e,t
heor
eti
cal
l
yanel
ect
ronmi
croscopecanr
esol
veobj
ect
swi
thadi
amet
erof
about0.
025Å(
i.
e.½*
0.05Å)
.Not
ethathy
drogenat
om hasadi
amet
erofabout1.
06Å
whi
ch i
slar
gert
han t
he di
amet
erofan obj
ectt
hatan el
ect
ron mi
croscope can
t
heor
eti
cal
l
yresol
ve;butduet
odi
ff
icul
ti
esi
ntheconst
ruct
ionofobj
ect
ivest
heact
ual
r
esol
vi
ngpoweroft
hebestmoder
nel
ect
ronmi
croscopei
sabout10Å. 1mm =1000
mi
cronsor1mi
cron=1000mi
l
li
micr
ons,
1mi
l
li
micr
on=10Å

B.Thedept
hfi
eld

Dept
hoff
iel
d(DOFi
)ist
hear
eai
nfr
ontofandbehi
ndt
hespeci
ment
hatwi
l
lbei
n
accept
abl
efocus.TheDept
hofFi
eld(
DOFi
)ofal
ensi
sit
sabi
l
ityt
omai
ntai
nadesi
red
amountofi
magequal
i
ty(
spat
ialf
requencyataspeci
fi
edcont
rast
),wi
thoutr
efocusi
ng,
i
ftheobj
ecti
sposi
ti
onedcl
osert
oandf
art
herf
rom bestf
ocus.DOFial
soappl
i
est
o
obj
ect
swi
thcompl
exgeomet
ri
esorf
eat
uresofdi
ff
erenthei
ght
s.Asanobj
ecti
spl
aced
cl
oserorf
art
hert
hant
hesetf
ocusdi
stanceofal
ens,t
heobj
ectbl
ursandbot
h

27
r
esol
uti
onandcont
rastsuf
fer
.Becauseoft
his,DOFonl
ymakessensei
fiti
sdef
ined
wi
thanassoci
atedr
esol
uti
onandcont
rast
.

Dept
hoff
ocusi
sessent
ialt
hesameasdept
hoff
iel
dbutf
oronei
mpor
tantdi
ff
erence,
t
hatbei
ngmagni
fi
cat
ion(
M).Wi
thhi
ghermagni
fi
cat
iondept
hoff
iel
dbecomesshor
ter
,
howev
erhi
ghermagni
fi
cat
ion i
ncr
ease t
hedept
h off
ocusf
ort
he i
mage.Thi
sis
becauset
hemagni
fi
cat
ioni
sdonewi
thapr
oject
ionl
ens.

Fi
eldofv
iew(
FOV)i
showmuchofaspeci
meni
svi
sibl
eatanygi
vent
imei
nthel
ater
al
pl
ane(
i.
e.,
per
pendi
cul
art
otheopt
ical
axi
s).I
tcanal
sobet
houghtofast
hedi
amet
erof
t
he ci
rcl
e ofl
i
ghtv
isi
ble when l
ooki
ng t
hrough a mi
croscope.FOV i
sinv
ersel
y
pr
opor
ti
onalt
othemagni
fi
cat
ion(
ast
hemagni
fi
cat
ioni
ncr
eases,t
heFOVdecr
eases)
.
Anot
herwayt
ounder
standt
hisi
stoconsi
dert
hatwhenaspeci
meni
smagni
fi
ed,t
he
mi
croscopei
szoomi
ngi
noni
tand,
consequent
ly,
seei
ngl
essofi
t(buti
ngr
eat
erdet
ail
).
Thesi
zeoft
heFOVort
hedi
amet
eroft
hev
isi
bleci
rcl
eofl
i
ght(
alsoknownast
heFi
eld
Si
ze)i
sequalt
otheFi
eldNumber(
FN)oft
heey
epi
ecedi
vi
dedbyt
hemagni
fi
cat
ionof
t
heobj
ect
ivel
ens(
measur
edi
nmi
l
li
met
ers)
.

C.I
magef
ormat
ion
The compound mi
croscope consi
sts oft
wo opt
icalcomponent
s(t
hus t
he t
erm
compound)
:theobj
ect
ivel
enssy
stem,whi
chhasav
eryshor
tfocaldi
stanceandi
s
pl
acedv
erycl
oset
otheobj
ect
;andt
heocul
arorey
epi
ecesy
stem,whi
chhasal
onger
f
ocall
engt
h,l
owermagni
fi
cat
ion;andwhi
chf
urt
hermagni
fi
esandpr
oject
sthei
mage
ont
other
eti
naoft
heey
e.Theobj
ect
ivel
enspr
oject
sar
eali
mage(
thei
nter
medi
ate
i
mage)oft
heobj
ectupi
nthebodyoft
hemi
croscopeont
otheobj
ect
iveconj
ugat
e
i
magepl
aneorpr
imar
yimagepl
ane.Theobj
ect
ivel
ensf
ormsar
eali
magei
nthe
mi
croscopebodyt
hatact
sast
heobj
ectf
ort
heocul
arl
ens.Ther
eali
magebecomes
t
heobj
ectf
ort
heey
eit
sel
fandi
spr
oject
edont
other
eti
na.Theocul
ari
ntur
nact
sasa
si
mpl
e magni
fi
ert
ofor
m al
arge v
irt
uali
mage att
hedi
stance ofmostdi
sti
nct
v
isi
on(
25cm)f
rom t
heey
e.

28
 Fi
gur
e3.
1.I
magef
ormat
ionwi
tht
hecont
extofar
eal
micr
oscope

Obj
ect
ives:
Att
heendoft
hissect
iont
het
rai
neeswi
l
lbeabl
eto:
 bet
weenmagni
fi
cat
ionandr
esol
uti
on.
 est
imat
ethesi
zeofi
mageseenunderl
i
ghtmi
croscope
Mat
eri
alsandchemi
cal
s:Mi
croscope,
micr
oscopesl
i
de,
cov
ersl
i
p,penci
l
,api
eceof
paper
,gr
aphpaper
,wat
er

D.Focusi
ng
1.St
udyofadot
Youar
epr
ovi
dedwi
thapi
eceofpaperbear
ingat
ypeddot(
ful
lst
op)
.Ifnot
,usi
nga
penci
lmakeasmal
ldotonasmal
lpi
eceofpaper
.
1.Wetmountt
hedot
2.St
udyt
hedotundert
hel
owpowerandt
henundert
hemi
ddl
epowerf
oll
owi
ngt
he
f
ocusi
ngst
epsont
hepr
evi
ouspages.
(
i) Underwhi
choft
hesepower
sdoest
hedotl
ookmor
edi
ff
use(
loosel
yhel
d

29
 t
oget
her
)
(
ii
)Whydoest
hedotl
ookdi
ff
usedundert
hemi
croscopet
hanwheny
oul
ookati
t
wi
tht
heunai
dedey
e?

E.Ty
peofi
magef
ormedandt
hedi
rect
ionofi
magemov
ement

1.Ty
peofi
magef
ormed

Youar
epr
ovi
dedwi
thasmal
lpi
eceofpaperwi
tht
hel
ett
er‘
e’t
ypedoni
t.Wetmounti
t
andobser
vei
tundert
hel
owpowerobj
ect
ive.
(
i) I
sthel
ett
eri
nver
tedf
rom t
opt
obot
tom?

(
ii
)Isi
tinv
ert
edf
rom l
eftt
ori
ght
?
(
ii
i)I
fyouwer
etomountt
hel
ett
erp,
whatt
hei
magewoul
dlookl
i
kei
fseenundert
he
mi
croscope
(
iv)I
ftheabov
eobj
ectst
andsi
nfr
ontofami
rr
orshowwi
thadr
awi
ngwhati
tsmi
rr
or
i
magewoul
dlookl
i
ke.
(
v) Fr
om y
ourabov
e obser
vat
ions how does t
he i
mage ofan obj
ectundera
mi
croscopedi
ff
erf
rom i
tsmi
rr
ori
mage?

2.Di
rect
ionofi
magemov
ement

(
i) Looki
ngi
ntot
hemi
croscopemov
ethesl
i
det
owar
dst
hel
eft
.Whi
chwaydoes
t
hei
magemov
e?
(
ii
)Ify
oumov
ethesl
i
deawayf
rom y
ouwhi
chwaydoest
hei
magemov
e?

F.Howt
ocal
cul
atemagni
fi
cat
ionandar
eaoff
iel
dofv
isi
on

Tot
almagni
fi
cat
ion
I
nacompoundmi
croscopet
hei
mageofanobj
ectt
hati
smagni
fi
edbyt
heobj
ect
ive
i
sagai
nmagni
fi
edbyt
heocul
ar.Ther
efor
e,t
het
otalmagni
fi
cat
ionpr
ovi
dedbya
compoundmi
croscopei
sthepr
oductoft
hemagni
fyi
ngpoweroft
heocul
arandt
he
par
ti
cul
arobj
ect
iveused.

i
. Whati
sthemagni
fi
cat
ionoft
hemi
croscopewhenusi
ngt
hel
ow power
?The

30
 medi
um power
?Thehi
ghpower
?
i
i
.Ift
heocul
arofy
ourmi
croscopei
smar
ked7.
5X,byhowmanydi
amet
erswi
l
lan
obj
ectv
iewedundermi
ddl
epowerbemagni
fi
ed?
i
i
i.I
fanobj
ectl
ooks2mm l
ongundert
hemi
ddl
epower
,whati
sit
sreal
lengt
h?
i
v.I
fanobj
ecti
s0.
001mm i
ndi
amet
erhowbi
gwoul
ditappeart
hehi
ghpowerof
y
ourmi
croscope?

Ar
eaoff
iel
dofv
isi
on

Thef
iel
dofv
isi
oni
sthear
eav
isi
blewheny
oul
ookt
hrought
hemi
croscope.Knowi
ng
t
hesi
zeoft
hef
iel
dofv
iewwi
l
lenabl
eyout
odet
ermi
net
hesi
zeoft
heobj
ecty
ouar
e
obser
ving.Speci
alr
uler
sar
eusedt
odet
ermi
net
hef
iel
dofv
iew andmeasur
eobj
ect
s
undert
hemi
croscope.

i
.Wetmountapi
eceofgr
aphpaperonagl
asssl
i
deandcountt
henumberof
mi
l
li
met
ersquar
esv
isi
bleunderl
ow powerandmi
ddl
epower
.Hal
formor
ethan
hal
fsquar
esar
ecount
edasf
ullsquar
esandt
hosel
esst
hanhal
fshoul
dnotbe
count
ed.

i
i
.Det
ermi
net
her
ati
ooft
hear
easoft
het
wof
iel
dsofv
isi
on(
underl
owpowerand
mi
ddl
epower
).

i
i
i.Howi
sfi
eldofv
isi
onr
elat
edt
omagni
fi
cat
ion?
i
v.Underwhi
chpower(
thel
ow ort
hemi
ddl
e)cany
ouseemor
epar
ts(
area)ofan
obj
ectatat
ime?
v
.Count
ingt
hesquar
esacr
osst
hemi
ddl
eoft
hef
iel
d ofv
isi
on,det
ermi
net
he
di
amet
ersoft
hel
owandmi
ddl
epowerf
iel
ds.Gi
vey
ourr
esul
tinmi
crons?
v
i.Whati
sther
ati
ooft
hedi
amet
eroft
hel
ow powerf
iel
dofv
isi
ont
othatoft
he
medi
um power
?
v
ii
.Fr
om t
her
ati
ooft
het
wodi
amet
ers,
det
ermi
net
her
ati
ooft
hemagni
fyi
ngpower
s
oft
hel
owandmi
ddl
epowerobj
ect
ives?
v
ii
i.Usi
ngt
hegr
aphpapercany
oudet
ermi
net
hedi
amet
eroft
hef
iel
dofv
isi
onoft
he
hi
ghpower
?Ifno,whynot
?Ify
es,gi
vet
hef
igur
esi
nmi
crons.Remembert
haty
ou

31
 shoul
dfocust
hehi
ghpowerwi
thf
ineadj
ust
mentonl
y!

D.Mi
croscopi
cexami
nat
ionofl
iv
ingor
gani
sms
A consi
der
abl
e amount of i
nfor
mat
ion can be gai
ned by car
efulmi
croscopi
c
exami
nat
ionofmi
croor
gani
sms.Li
vi
ngor
gani
smscanbest
udi
edt
odet
ermi
net
he
nat
uralsi
zeandshapeofcel
l
s,cel
l
ularar
rangement
,andmot
il
it
y,r
eact
ionst
ovar
ious
chemi
cal
sori
mmuneser
aandr
esponset
oenv
ironment
alf
act
ors.

Di
rectexami
nat
ionofl
i
vingor
gani
smsi
ncl
udi
ngbact
eri
a,pr
otozoanandot
her
scanbe
st
udi
edbyt
womeans:WetmountandHangi
ngdr
opt
echni
que.Bot
har
ever
yusef
uli
n
det
ermi
ningsi
ze,
shapeandmov
ement
.
A wetmounti
saf
ast
erwayt
oobser
vebact
eri
a.I
nHangi
ngdr
opt
echni
que,t
he
appl
i
cat
ionofpet
rol
eum sealar
oundt
hecav
ityr
educest
heev
apor
ati
onofsuspended
f
lui
ddr
op.I
tal
somakespossi
blet
oobser
vel
argermi
crobesandmot
il
eor
gani
sms
mor
eeasi
l
ybecauseofgr
eat
erdept
hpr
ovi
dedbyt
hehangi
ngdr
op.

Mat
eri
alsandchemi
cal
s:Mi
croscopi
csl
i
des,
cov
ersl
i
ps,
hangi
ngdr
op(
cav
ity
)sl
i
de,
i
nocul
ati
ngl
oop,
toot
hpi
ck,
spr
itl
amp,
brot
hcul
tur
e,pet
rol
eum j
ell
y(Vasel
i
ne)
,di
sti
l
led
wat
eri
nadr
opper
,oi
li
mmer
sion

1.Wetmountt
echni
que
Pr
ocedur
e:
1.Transf
erasmal
ldr
opoft
hebact
eri
alcul
tur
egi
venont
hecent
erofcl
eangl
ass
sl
i
des.
2.Handl
ethecov
ersl
i
pbyi
tsedgesandpl
acei
tont
hedr
op
3.Pr
esst
hecov
ersl
i
pgent
lywi
tht
heendofapenci
l

2.Hangi
ngdr
opt
echni
que
Pr
ocedur
e:
1.Wi
that
oot
hpi
ck,
spr
eadasmal
lri
ngofVasel
i
near
oundt
heconcav
ityofa
depr
essi
onsl
i
de.Donotuset
oomuchVasel
i
ne.
2. Use i
nocul
ati
ng l
oop t
o asept
ical
l
y pl
ace a smal
ldr
op oft
he bact
eri
al

32
 suspensi
oni
nthecent
erofacov
ersl
i
p.
3. Lowert
hedepr
essi
onsl
i
de,wi
tht
heconcav
ityf
aci
ngdown,ont
othecov
ersl
i
p
sot
hatt
hedr
oppr
otr
udesi
ntot
hecent
eroft
heconcav
ityoft
hesl
i
de.Pr
ess
gent
lyt
ofor
m aseal
4. Tur
nthehangi
ngdr
opsl
i
deov
erandpl
aceont
hest
ageoft
hemi
croscopeso
t
hatt
hedr
opi
sov
ert
hel
i
ghthol
e.
5. Ex
ami
net
hedr
opbyf
ir
stl
ocat
ingi
tsedgeunderl
owpowerandf
ocusi
ngont
he
dr
op.Swi
tcht
othehi
gh‐
powerobj
ect
iveandt
hen,usei
mmer
sionoi
lobj
ect
ive.
Becar
efult
odi
sti
ngui
shbet
weenmot
il
it
yandBr
owni
anmov
ement
.Br
owni
an
mov
ementr
esul
tsf
rom t
her
andom mot
ionofwat
ermol
ecul
esbombar
dingt
he
bact
eri
acausi
ngt
hem t
omov
e

Quest
ions:
1.Whyar
emi
croor
gani
smsdi
ff
icul
ttoseei
nwetpr
epar
ati
ons?
2.Whati
stheadv
ant
ageofahangi
ng-
droppr
epar
ati
onov
ert
hatofwetmount
pr
epar
ati
on?
3.Di
sti
ngui
shbet
ween‘
tr
uemot
il
it
y’and‘
Browni
anmov
ement
’?

33
 LABORATORYSTUDYNUMBER3

THECELLANDCELLSTRUCTURE

3.
1Int
roduct
ion
Thecel
li
sthest
ruct
ural
andf
unct
ional
uni
tofl
i
fe.Ont
hebasi
sofl
evel
ofcel
l
ular
or
gani
zat
iont
wot
ypesofcel
l
sar
erecogni
zed.Theyar
epr
okar
yot
iccel
ls(
Prokar
yot
ic
cel
l
sar
efoundi
nbact
eri
a,i
ncl
udi
ngbot
hAr
chaebact
eri
aandEubact
eri
a,andi
ncl
udi
ng
t
hebl
ue-
greenal
gae)andeukar
yot
iccel
ls(
aref
oundi
nani
mal
s,pl
ant
s,Fungi
and
pr
oti
sts)
.Thel
att
erar
echar
act
eri
zedbyt
hecompar
tment
ali
zat
ionofi
tspar
tsi
.e.i
ts
par
tsar
edi
vi
dedi
ntosepar
atecompar
tment
ssuchasnucl
eus,
mit
ochondr
ia,
etc.

Thepr
otopl
asm ofaeukar
yot
iccel
lisdi
vi
dedi
ntonucl
eusandcy
topl
asm.Thenucl
eus
i
sencl
osedbyanucl
earmembr
aneconsi
sti
ngoft
woconcent
ri
cmembr
anes.The
nucl
earmembr
anei
sper
for
atedbyanumberofnucl
earpor
es(
Annul
i)
.Thepr
omi
nent
bodyi
nthenucl
eusi
sthenucl
eol
us.Ther
ecoul
dbemor
ethanonenucl
eol
ii
nanucl
eus.
Mostoft
hespacei
nthenucl
eusi
soccupi
edbyt
hint
hreadl
i
kebodi
escal
l
edchr
omat
in.
Thechr
omat
ini
sthemat
eri
aloft
hechr
omosomeandi
tconsi
stsofDNAandspeci
al
pr
otei
ns compl
exed wi
tht
he DNA.The gr
ound mat
eri
ali
nthe nucl
eus i
s cal
l
ed
nucl
eopl
asm.

I
nthecy
topl
asm t
her
ear
enumer
ousbodi
esofv
ari
oussi
zes,shapeandf
unct
ion;
net
wor
ksofi
nter
connect
ingt
ubul
es(
channel
s)andv
ari
ousmembr
anesboundv
esi
cles
andv
acuol
es.

34
Amongt
hebodi
esi
nthecy
topl
asm ar
ethemi
tochondr
ia(
thepowerhouseoft
hecel
l
)
andt
her
ibosomes.Ther
ibosomesar
esi
tesofpr
otei
nsy
nthesi
si.
e.‘
benches’upon
whi
chami
noaci
dsar
eassembl
edi
ntopr
otei
n.Ther
ibosomesar
efoundei
therf
reel
yin
t
hecy
topl
asm orat
tachedt
othesur
facesofmembr
aneouschannel
s.

The i
nter
connect
ing t
ubul
es (
channel
s) i
nthe cy
topl
asm f
orm what i
s cal
l
ed
endopl
asmi
cret
icul
um.Theendopl
asmi
cret
icul
um and t
hatwhi
ch i
sfr
eeoft
he
r
ibosomesi
sknownassmoot
hendopl
asmi
cret
icul
um.I
nthecy
topl
asm t
her
eisal
so
anot
herv
esi
cul
arst
ruct
ureknownast
he(
Gol
gibodyorGol
giappar
atus)namedaf
ter
Cami
l
loGol
gi,whof
ir
stdescr
ibedi
t.Thi
sbodyappear
sasacol
l
ect
ionofcl
osel
y
at
tachedst
ackoff
lat
tenedsacs.

I
nthecy
topl
asm t
her
ear
eal
soenzy
me-
cont
aini
ngv
esi
clescal
l
edLy
sosomesand
Per
oxi
somes.Thef
ormercont
ainv
ari
ousdi
gest
iveenzy
mesandt
hel
att
ercont
ain
enzy
mesr
esponsi
blef
ort
hepr
oduct
ionordegr
adat
ionofpr
oxi
dessuchast
heenzy
me
cat
alaset
hatdegr
adeshy
drogenper
oxi
de(
H2O2)t
owat
erandoxy
gen.
Somev
esi
clesar
econcer
nedwi
tht
ranspor
tat
ionofmat
eri
alsi
ntoandoutoft
hecel
l
.
Endocy
ticVesi
clesar
efor
medf
rom t
hecel
lmembr
aneandmov
emat
eri
alsi
ntot
hecel
l
.
Ther
ear
etwo t
ypes-pi
nocy
ticv
esi
cles(
transpor
tli
qui
d)and phagocy
ticv
esi
cles
(
transpor
tsol
i
dmat
eri
al)
.Exocy
ticv
esi
clesbudoft
heGol
giappar
atusandt
ranspor
t
cel
lsecr
eti
ont
oout
side.

Ani
malcel
l
scapabl
e ofdi
vi
sion hav
e neart
he nucl
eusatl
easta pai
rofhal
l
ow
cy
li
ndr
icalbodi
escal
l
edcent
ri
oles.Ther
ear
eal
sof
inet
ubul
esandf
il
ament
sknownas
mi
crot
ubul
esandmi
crof
il
ament
srespect
ivel
y.

Thegr
oundsubst
ancei
nwhi
cht
hev
ari
ousor
ganel
l
esoft
hecel
lar
esuspendedi
s
cal
l
ed cy
tosol
,lar
gel
ycomposed ofwat
eri
n whi
ch v
ari
ousi
nor
gani
cand or
gani
c
subst
ances ar
e di
ssol
ved orsuspended.The whol
e cel
lis encl
osed by t
he cel
l
membr
ane.

I
naddi
ti
ont
omostoft
hest
ruct
ureshowni
ntheani
malcel
l
,pl
antcel
l
shav
ecer
tai
n
st
ruct
urest
hatar
enotf
oundi
nani
malcel
l
s.Thei
mpor
tantonesar
epl
ast
ids(
e.g.
chl
oropl
ast
s),cel
lwal
landal
argecent
ralv
acuol
ewhi
choccupi
esasmuchas80-
90%

35
oft
het
otal
cel
lvol
umeofamat
urecel
l
.

Fi
gur
e3.
1:Thest
ruct
ureofat
ypi
cal
plantcel
lasr
eveal
edbyel
ect
ronmi
croscope

36
Fi
gur
e3.
2:Thest
ruct
ureofat
ypi
cal
ani
mal
cel
lasr
eveal
edbyel
ect
ronmi
croscope

ASTUDYOFPLANTANDANI
MALCELLS
Obj
ect
ives:
Af
tercompl
eti
ngt
heact
ivi
tyt
rai
neeswi
l
lbeabl
eto:
 mountpl
antcel
l
sandani
mal
cel
l
sandobser
vet
hemaj
orv
isi
blest
ruct
ures
 di
sti
ngui
shbet
weenpl
antcel
l
sandani
mal
cel
l
s.
Act
ivi
ty1:Obser
vingpl
antcel
ls(
oni
onepi
der
malcel
ls)
Anoni
onbul
bismadeupofsev
erall
ayer
sofmodi
fi
edl
eav
es.Eachl
eafhasdel
i
cat
e
epi
der
mall
ayer
swhi
chcaneasi
l
ybepeel
edof
f.Theepi
der
mall
ayerconsi
stsofcel
l
s
t
hatoni
onl
eafcanber
eadi
l
yseenundert
hemi
croscope.

37
Mat
eri
als:Mi
croscope,
micr
oscopesl
i
des(
depr
essi
onandf
lat
),cov
ersl
i
ps,
paper
t
owel
i
ng,
toot
hpi
cks(
roundend)
,ty
pical
ani
mal
cel
lsl
i
de,
needl
e
Chemi
cal
s:Al
cohol
,i
odi
ne,
JanusGr
eenBst
ain,
dist
il
ledwat
er
Pr
ocedur
e:
1.Peel
thewhi
ti
shi
nnerepi
der
misf
rom oneoft
hel
eav
es.
2.Pl
acet
hepi
eceoft
heepi
der
misonagl
asssl
i
de,
addadr
opofwat
erandt
hena
cov
ersl
i
p(Not
ethatwheny
oucov
ert
hecov
ersl
i
p,t
ryt
oav
oidai
rbubbl
est
hat
maymi
stakey
ouwi
thcel
lor
ganel
l
es.Youri
nst
ruct
orshowsy
ouhowt
oav
oidai
r
bubbl
es)
.
3.Exami
nei
tundert
hemi
ddl
epower
.
a.I
sthenucl
euscl
ear
lyv
isi
ble?
b.I
sthecy
topl
asm t
hickeri
nthecent
reori
ntheper
ipher
yoft
hecel
l
?
4.Remov
ethesl
i
def
rom t
hest
ageandpl
aceadr
opofi
odi
nesol
uti
onatoneend,
hol
d
api
eceofabsor
bentpaperatt
heopposi
teendoft
hecov
ersl
i
p.Wai
tforabout5
mi
nut
est
il
lthecel
l
sgetst
ained.(
N.B.i
fyourmounti
snotgoody
oucanmountf
resh
pi
eceofepi
der
misdi
rect
lyi
nadr
opofi
odi
ne)
.

a.Exami
neundert
hemi
ddl
epoweranddr
awt
woormor
eadj
acentcel
l
sandl
abel
t
hepar
ts.
b.Whi
chpar
t(s)oft
hecel
lbecamemor
edeepl
yst
ained?
c.Howdoy
ouaccountf
ort
hedi
ff
erencei
nst
aini
ng?

Act
ivi
ty2:Obser
vingani
mal
cel
ls(
cheekcel
ls)
Mat
eri
als:
Micr
oscope,
micr
oscopesl
i
des,
cov
ersl
i
ps,
toot
hpi
ck,
dropper
,
Chemi
cal
s:Met
hyl
enebl
ue

Pr
ocedur
e:

1.Gent
lyscr
apet
hei
nsi
deofy
ourcheekort
hei
nsi
deofy
ourl
owerl
i
pwi
tht
hebr
oad
endofacl
eant
oot
hpi
ck
2.St
irt
hescr
api
ngi
ntoadr
opofwat
eronacl
eangl
asssl
i
de.
3.Pl
aceacov
ersl
i
poni
tandexami
nei
tundert
hemi
ddl
epower
.

38
 a.Ar
ethepar
tsoft
hecl
ear
lyv
isi
ble?
b.Whati
sthecol
oroft
hecel
l
?
4.Pl
aceadr
opofmet
hyl
enebl
ueatoneendandhol
dapi
eceofabsor
bentpaperatt
he
opposi
te endsoft
hecov
ersl
i
p.Wai
tforabout5mi
nut
esunt
ilt
hecel
l
sgetst
ained.
5.Exami
ney
ourpr
epar
ati
onundert
hemi
ddl
epower(
ory
oucanuset
hehi
ghpower
wi
thgood car
e).
a.Cany
ounowdi
sti
ngui
sht
henucl
eusandt
hecy
topl
asm?
b.Compar
eposi
ti
onoft
henucl
eusi
ncheekcel
l(ani
malcel
l
)wi
tht
hatofoni
oncel
l
(
plantcel
l
).
c.Dr
awt
woormor
eadj
acentcel
l
sandl
abel
thepar
t

39
 LABORATORYSTUDYNUMBER4

DI
FFUSI
ONANDOSMOSI
S

4.
1Int
roduct
ion
Ther
andom mov
ementofmol
ecul
esf
rom ar
egi
onofhi
gherconcent
rat
iont
oar
egi
on
ofl
owerconcent
rat
ioni
sknownasdi
ff
usi
on.I
ttendst
odi
str
ibut
emol
ecul
esuni
for
mly
.
Br
owni
anmot
ion(
orBr
owni
anmov
ement
)canbedef
i t
nedas"her
andom movementof
mi
croscopi
cpar
ti
clessuspendedi
naf
lui
d."
Osmosi
sist
hedi
ff
usi
on(
netmov
ement
)ofsol
ventpar
ti
cles(
mol
ecul
es)acr
ossasemi
-
per
meabl
emembr
anef
rom ar
egi
onoft
hei
rhi
gherconcent
rat
ion(
i.
e.f
rom week
sol
uti
on)t
oar
egi
onoft
hei
rlowerconcent
rat
ion(
i.
e.t
ost
rongsol
uti
on)
.Inbi
ologi
cal
sy
stemssi
ncet
hesol
venti
swat
erandt
hemembr
anei
sthecel
lmembr
ane,osmosi
s
canbedef
inedast
hedi
ff
usi
onofwat
ermol
ecul
esacr
osst
hecel
lmembr
anef
rom a
r
egi
onofhi
gherconcent
rat
iont
oar
egi
onoft
hei
rlowerconcent
rat
ion.

The netmov
ementofwat
eri
nto oroutoft
he cel
ldepends upon t
he r
elat
ive
concent
rat
ionoft
hepr
otopl
asm andt
hesol
uti
onsur
roundi
ngt
hecel
l
.Whenosmosi
s
r
esul
tsi
nthenetmov
ementoft
hewat
eroutofacel
l
,thepr
otopl
asm oft
hecel
lshr
inks.
Thi
sshr
inkageoft
hepr
otopl
asm asar
esul
tofnetl
ossofwat
eri
scal
l
edpl
asmol
ysi
s.
Theshr
inkageofani
mal
cel
li
scal
l
edcr
enat
ion.
Whenosmosi
sresul
tsi
n,anetmov
ementofwat
eri
ntoacel
l
,thepr
otopl
asm swel
l
s.I
f
i
tisapl
antcel
l
,theswel
l
ingpr
otopl
asm exer
tsapr
essur
e(t
urgorpr
essur
e)agai
nstt
he
cel
lwal
l
.Ev
ent
ual
l
ythenetmov
ementoft
hewat
eri
ntot
hecel
lisst
oppedwhent
he
pr
essur
e exer
ted i
nwar
ds byt
he wal
land t
he pr
essur
e exer
ted out
war
ds byt
he
pr
otopl
asm i
sequal
i
zed.Anani
malcel
l(e.
g.bl
oodcel
l
)si
ncei
tdoesnothav
ear
igi
d
cel
lwal
l
;itmaybur
sti
fitt
akesi
nexcesswat
er.

Obj
ect
ives:
Af
tercompl
eti
ngt
heact
ivi
tyst
udent
swi
l
lbeabl
eto:
 descr
ibet
hesi
gni
fi
canceofdi
ff
usi
onandosmosi
sinl
i
vingor
gani
sms

40
  expl
ainwhydi
ff
erentsubst
anceswi
thdi
ff
erentmol
ecul
arwei
ght
sdi
ff
eri
nthei
r
r
ateofdi
ff
usi
on

I
.Di
ff
usi
on
Mat
eri
als:Pet
ridi
shcont
aini
ngsol
i
dif
iedagar
,Cor
kbor
er,
beaker
s
Chemi
cal
s:Pot
assi
um per
manganat
e,met
hyl
enebl
ue

Act
ivi
ty1:Det
ermi
nat
ionoft
her
ateofdi
ff
usi
on
1.Youar
epr
ovi
ded(
ingr
oups)wi
thaPet
ridi
shcont
aini
ngsol
i
dif
iedagar
.
2.Usi
nganumber5Cor
kbor
er,
punchf
ourhol
esi
ntheagar
3.Fi
l
loneoft
hehol
eswi
thoneort
wodr
opsofpot
assi
um per
manganat
e(mol
ecul
ar
wei
ght=158.
05)andt
heot
heronewi
thmet
hyl
enebl
ue(
mol
ecul
arwei
ght=
319.
86)
.
4.Per
iodi
cal
l
yexami
net
hePet
ridi
shandr
ecor
dyourf
inal
obser
vat
ion.
a.Whi
chchemi
cal
dif
fusedf
ast
er?
b.Whi
chchemi
cal
trav
elst
heshor
testdi
stance?
c.How do y
ou accountf
ort
he di
ff
erence i
nthe di
ff
usi
on r
ates oft
he
chemi
cal
s?

Act
ivi
ty2:
Dif
fusi
onexper
imentbyusi
ngpl
ast
icbagasr
epr
esent
ingcel
lmembr
ane
Mat
eri
als
•400-
mLbeaker •t
wistt
ie
•25-
mLgr
aduat
edcy
li
nder •i
odi
nesol
uti
on
•1%st
archsol
uti
on •f
orceps
•Pl
ast
icsandwi
chbag
Pr
ecaut
ion:I
odi
nesol
uti
oncani
rr
it
atet
heey
esandski
nandcanst
aincl
othi
ng.Wear
saf
etygoggl
es,
glov
es,
andal
abor
ator
yapr
onwhi
l
ehandl
i
nganysol
uti
ont
hatcont
ains
i
odi
ne.Ri
nseof
fanysol
uti
ont
hatspi
l
lsony
ourski
norcl
othi
ng.Washy
ourhands
t
hor
oughl
ywi
thsoapandwar
m wat
erbef
orel
eav
ingt
hel
ab.
Pr
ocedur
e
1.Putony
ourgoggl
es,
apr
on,
andgl
oves.
41
2.Addabout200mLofwat
ert
oa400-
mLbeaker
.
3.Pour25mLofst
archsol
uti
oni
ntot
hepl
ast
icsandwi
chbag.
4.Useat
wistt
iet
oti
ght
lyseal
thebag.
5.Uset
apwat
ert
othor
oughl
yri
nset
heout
sideoft
hebagi
ncaseanyst
archsol
uti
on
spi
l
ledont
otheout
sideoft
hebag.Besur
etor
inset
het
wistt
ieaswel
l
.
6.Pl
acet
hepl
ast
icbagi
nthebeakersot
hatt
hebagi
scompl
etel
ycov
eredwi
thwat
er.
7.Add 8 dr
opsofi
odi
nesol
uti
on t
othewat
eri
nthebeaker
.Recor
dyouri
nit
ial
obser
vat
ionsi
nthedat
atabl
e.Wai
t10mi
nut
es,
andt
henr
ecor
dyourf
inal
obser
vat
ions.
8.Useaf
orcepst
oremov
eanddi
sposeoft
hepl
ast
icbagasi
nst
ruct
edbyy
ourt
eacher
.
Quest
ions:
1.Af
tery
oupl
acedt
hepl
ast
icbagi
nthebeaker
,whathappenedt
othei
odi
ne?What
happenedt
othest
arch?
2.Usewhaty
ouknowaboutt
hest
ruct
ureofst
archmol
ecul
est
oexpl
ainy
ourr
esul
ts.
3.Di
dwat
ermov
eint
othebagoroutoft
hebag?Why
?

I
I.Osmosi
s
Act
ivi
ty1:St
udy
ingosmosi
susi
ngoni
onepi
der
mis
Mat
eri
als:
Micr
oscope,
micr
oscopesl
i
de,
cov
ersl
i
p,oni
onr
oot
,needl
e,dr
opper
Chemi
cal
s:1NNaCl
sol
uti
on,
wat
er
Pr
ocedur
e:
1.Remov
easmal
lpi
eceofoni
onepi
der
misandmounti
tinwat
er.Theout
er,
pi
nki
shepi
der
mismaybepr
efer
redsi
ncet
hecy
topl
asm caneasi
l
ybeseen
undert
hemi
croscope
2.Obser
vey
ourpr
epar
ati
onundert
hemi
ddl
epoweranddr
aw2or3adj
acentcel
l
s
andl
abel
thepar
ts.
3.Usi
ngacl
eansl
i
demountaf
reshepi
der
misi
nadr
opof1N NaCl(
inst
eadof
wat
er,
useNaCl
forwetmount
ing)
.Obser
vei
tundert
hemi
ddl
epower
.
4.Af
terabout20mi
nut
esobser
vei
tagai
nanddr
aw2or3adj
acentcel
l
sandl
abel
t
hepar
ts.
Quest
ions:
a.Whatmaj
ordi
ff
erencesdoy
ouobser
vebet
weent
hecel
l
smount
edi
nwat
er

42
 andt
hosemount
edi
nNaCl
sol
uti
on?
b.Expl
ainhowt
hedi
ff
erencecoul
dcomeabout
.

Act
ivi
ty2:I
nvest
igat
ingosmosi
susi
ngpot
atocy
li
nder
s/st
ri
ps
Osmosi
scanbeeasi
l
ydemonst
rat
edi
nbi
ologi
cal
syst
emsusi
ngpot
atost
ri
ps,
wat
er
andsal
torsugarsol
uti
on.
Mat
eri
als:
Pot
atot
uber
,cor
kbor
er,
beaker
s,r
uler
,el
ect
roni
cbal
ance,
tissuepaper
Chemi
cal
s:Sucr
ose,
dist
il
ledwat
er

Pr
ocedur
e:
1.Usi
ngnumber4cor
kbor
er,
take7pot
atocy
li
nder
sfr
om af
reshpot
atot
uber
.
2.Cutt
hepot
atocy
li
nder
seach4cm l
ongandwei
ght
hem separ
atel
y,r
ecor
dingt
he
i
nit
ial
massandl
engt
h.
3.Obser
veeachst
ri
pbyf
eel
i
ngi
twi
thy
ourf
inger
s,not
ingwhet
heri
tist
urgi
dorf
lacci
d.
Recor
dthi
s.
4.Takesi
xbeaker
sof50ml
vol
umeandl
abel
them as1-7.
5.Pr
epar
e2,
4,6,
8,10,
and15%sucr
osesol
uti
onssepar
atel
y.
6.Add30mlofdi
sti
l
ledwat
ert
obeaker1,andadd30mlofeachoft
hesucr
ose
sol
uti
onssequent
ial
l
ytobeaker
s2-7.
7.Pl
aceast
ri
pineachbeaker
.Recor
dthet
ime.
8.Af
ter3hour
sremov
eeachst
ri
panddabi
tont
issuepaper
.Takecar
eofnotmi
xi
ng
t
hem.
9.Wei
ghandmeasur
eeachpot
atost
ri
p,r
ecor
dingt
hef
inal
massandl
engt
h.
10.
Obser
veeachst
ri
pbyf
eel
i
ngi
t,not
ingwhet
heri
tist
urgi
dorf
lacci
d.Recor
dthi
s.
11.
Per
for
m %di
ff
erencecal
cul
ati
onsf
ort
hemassandl
engt
husi
ngt
hef
ormul
a:
Fi
nal-I
nit
ial
x100%
Ini
ti
al

Quest
ions:
1.I
nwhi
chpot
atocy
li
nder
shav
eyouobser
vedchangesi
nlengt
handmass?
Expl
ainr
easonsf
ort
hechange.
2.Whi
chpot
atocy
li
nder
shav
ebecomet
urgi
d?Whi
choneshav
ebecomef
lacci
d?

43
 Whi
chonesdoy
out
hinkhav
enei
therf
lacci
dnort
urgi
d?Expl
ainy
ourr
esul
tsbr
ief
ly.
3.Whati
sthesemi
per
meabl
emembr
anei
nthecel
l
soft
hepot
ato?

44
 LABORATORYSTUDYNUMBER5

TESTI
NGFORBI
OLOGI
CALLYI
MPORTANTMOLECULES

5.
1Int
roduct
ion

I
.Car
bohy
drat
es

Car
bohy
drat
esar
ecompoundsofcar
bon,hy
drogenandoxy
genoccur
ri
ngi
nav
eri
tyof
f
ormsi
nbot
hpl
ant
sandani
mal
s.Theyi
ncl
udesuchsubst
anceassugar
,st
arch,
gl
ycogenandcel
l
ulose.Car
bohy
drat
esser
veasst
ruct
uralsubst
ance.(
E.g.Cel
l
ulosei
n
pl
antcel
lwal
l
),chi
ti
ninsomef
ungi
,mur
inst
rengt
henmat
eri
alsofbact
eri
alcel
lwal
l
.As
wel
lasi
mmedi
atesour
ceofpot
ent
ial
ener
gyr
eser
ve(
E.g.Gl
ucose)
.
On t
he basi
s ofsi
ze (
i.
e.,numberofsugaruni
tst
hatt
hey ar
e composed of
),
car
bohy
drat
esar
eknown asmonosacchar
ide,di
sacchar
ides,ol
i
gosacchar
idesand
pol
ysacchar
ides.
Gl
ucose+GlucoseMaltose(mal tsugar
)
Fr
uctose+GlucoseSucrose(canesugar)
Gal
actose+GlucoseLactose(milksugar)

45
Oxi
dat
ion/
Reduct
ion(
Redox)React
ions:Oxi
dat
ioni
sgener
all
yref
err
edt
oast
hel
ossof
el
ect
rons,
whi
l
ereduct
ioni
sthegai
nofel
ect
rons.

46
Benedi
ct’
sreact
ionf
orReduci
ngSugar
sisshownas:
Cu2++e–Cu+
Copper(
II
)ions(
Cu2+)ar
ereducedt
ocopper(
I)i
ons(
Cu+)bygai
ningel
ect
ronsf
rom
t
her
educi
ngsugar
s.Ther
educi
ngsugari
soxi
dizedasar
esul
tofgi
vi
ngupi
tsel
ect
ron.
I
tisof
tendi
ff
icul
ttoseewher
eel
ect
ronsar
efl
owi
ngwheny
oucan’
tseeani
onchar
ge.
Howev
er,si
ncet
hemov
ementofhy
dro-
genof
tenf
oll
owst
heel
ect
rons,wecoul
d
descr
ibeoxi
dat
ionast
hel
ossofhy
drogenandr
educt
ionast
hegai
nofhy
drogen.

Numer
oust
est
shav
ebeendev
isedf
ort
hedet
ermi
nat
ionoft
hepr
oper
ti
esandf
ort
he
det
ermi
nat
ionofcar
bohy
drat
es.Abr
iefdescr
ipt
ionoft
hemostcommont
est
sfol
l
ows.
Obj
ect
ives:
Af
tercompl
eti
ngt
heact
ivi
ti
est
rai
neeswi
l
lbeabl
eto:
 det
ermi
ne an unknown f
ood sampl
e(car
bohy
drat
e,pr
otei
n,or l
i
pid) by
conduct
ingf
oodt
est
.
 di
ff
erent
iat
ebet
weenr
educi
ngsugar
sandnon-
reduci
ngsugar
s.

I
. Test
sforcar
bohy
drat
es
Act
ivi
ty1:Mol
isch’
sTest
Mol
i
sch’
stesti
sacommont
estf
oral
lcar
bohy
drat
esl
argert
hant
etr
oses.Thet
esti
s
ont
hebasi
sthatpent
osesandhexosesar
edehy
drat
edbyconcent
rat
edsul
phur
icaci
d
t
ofor
mfur
fur
alorhy
droxy
met
hyl
fur
fur
al,
respect
ivel
y.Thesepr
oduct
scondensewi
thα-
napht
hol
tof
orm pur
plecondensat
ionpr
oduct
.

47
Mat
eri
als:
Testt
ubes,
testt
uber
ack,
testt
ubehol
der
Chemi
cal
s:Di
sti
l
led wat
er,Mol
i
sch’
sreagent
,concent
rat
ed sul
phur
ic aci
d,Al
pha-
napht
hol sol
uti
on
Pr
ocedur
e:
1.Add2dr
opsoft
heα-
napht
holsol
uti
on(
5%i
net
hanolpr
epar
efr
esh)t
o2mlof
t
estsol
uti
oni
nat
estt
ube.
2.Car
eful
l
y,pourabout1mlofconc.H2SO4 downt
hesi
deoft
het
ubesoast
o
f
ormt
wol
ayer
s.
3.Car
eful
l
yobser
veanycol
ourchangeatt
hej
unct
ionoft
het
wol
i
qui
ds.
4.Repeatt
het
est
,usi
ngwat
eri
nst
eadoft
hecar
bohy
drat
esol
uti
on.
a. Di
dyouobser
veanycol
or?
b. Whatcol
ordi
dyouobser
ve?
c. I
fther
eisanycol
orf
ormat
ion,
wher
edi
ditf
orm?

Reduci
ngandnon-
reduci
ngsugar
s
Somesugar
shav
ethecapaci
tyt
oreducecer
t ncompoundssuchasCu2+.Suchsugar
ai s
ar
ecal
l
edr
educi
ngsugar
s.Reduci
ngsugar
sincl
udeal
lmonosacchar
ides,suchas
gl
ucose,f
ruct
ose,andsomedi
sacchar
idessuchasmal
tose.Anon-
reduci
ngsugari
sa
car
bohy
drat
ethati
snotoxi
dizedbyaweakoxi
dizi
ngagent(
anoxi
dizi
ngagentt
hat
oxi
dizesal
dehy
desbutnotal
cohol
s,suchast
heTol
l
en’
sreagent
)inbasi
caqueous
sol
uti
on.Thechar
act
eri
sti
cpr
oper
tyofnon-
reduci
ngsugar
sist
hat
,inbasi
caqueous
medi
um,
theydonotgener
ateanycompoundscont
aini
nganal
dehy
degr
oup.
Exampl
e:sucr
ose,
whi
chcont
ainsnei
therahemi
acet
algr
oupnorahemi
ket
algr
oupand,
t
her
efor
e,i
sst
abl
einwat
er.

A.Testf
orr
educi
ngsugar
s
Benedi
ct’
stest
Whenasol
uti
oncont
aini
ngBenedi
ct’
sreagentandar
educi
ngsugarar
eheat
ed,t
he
copper(
II
)ionsi
nar
ereducedt
ocopper(
I)i
onsandt
hesol
uti
onchangesi
ncol
or.
Benedi
ct’
sreagentcont
ainspot
assi
um t
hiocy
anat
eandi
susedt
odet
ermi
nehowmuch
r
educi
ngsugari
spr
esent
.Thi
ssol
uti
onf
ormsacoppert
hiocy
anat
epr
eci
pit
atewhi
chi
s
whi
teandcanbeusedi
nat
it
rat
ion.
48
Mat
eri
als:
Testt
ubes,
testt
ubehol
der
,wat
erbat
h
Chemi
cal
s:5% ofgl
ucoseor5% off
ruct
oseor5% ofmal
tose,Benedi
ct’
ssol
uti
on,
di
sti
l
led wat
er

Pr
ocedur
e:

1.Add2ml
of0.
1-1%ofasol
uti
onoft
her
educi
ngsugart
oat
estt
ube.
2.Add2ml
ofdi
sti
l
ledwat
ert
oanot
hert
estt
ube.
3.Addanequal
(3ml
)vol
umeofBenedi
ct’
ssol
uti
ont
obot
htestt
ubes.
4.Shakeandboi
lthet
estt
ubei
nawat
erbat
hgent
ly.
a.Di
dyounot
eanycol
orchange?
b.I
fso,
whatcol
orchanges?
c.I
nwhi
cht
estt
ubedi
dyounot
ethecol
orchange?
d.Whati
sthesour
ceoft
hecol
orchange?

B. Testf
ornon-
reduci
ngsugar
s
Mat
eri
als:Testt
ubes,
testt
uber
ack,
testt
ubehol
der
,Bunsenbur
ner
Chemi
cal
s:5% sucr
osesol
uti
on,pH paper
/l
it
muspaper
,Benedi
ct’
ssol
uti
on,Sodi
um
hy
drogencar
bonat
e,Hy
drochl
ori
caci
d,di
sti
l
ledwat
er
Pr
ocedur
e
1.Add2ml
ofsucr
osesol
uti
ont
oat
estt
ube.
2.Add1ml
ofdi
l
utehy
drochl
ori
caci
d.
3.Boi
lforonemi
nut
e.
4.Car
eful
l
yneut
ral
i
zewi
thSodi
um hy
drogencar
bonat
e(checkwi
thpHpaper
).
5.Car
ryoutBenedi
ctst
est
.
a.Whatcol
orchangedi
dyouobser
ve?
b.Ar
ethecol
orchangesdi
ff
erentf
rom t
hepr
evi
ousexper
iment
?
c.Whati
sthepur
poseoft
hehy
drochl
ori
caci
d?
d.Whatst
atementy
oucanmakef
rom t
hisexper
iment
?

Testf
orst
arch(
Iodi
net
est
)
St
archi
sonl
ysl
i
ght
lysol
ubl
einwat
er,
inwhi
chi
tfor
msacol
l
oidalsuspensi
on.I
tcanbe

49
t
est
edi
nsuspensi
onorasasol
i
d.
Mat
eri
als:Testt
ubes,
testt
uber
ack,
Testt
ubehol
der
,Bunsenbur
ner
Chemi
cal
s:5% st
archsol
uti
on,gl
ucosesol
uti
on,sucr
ose,i
odi
ne/
pot
assi
um i
odi
de
sol
uti
on,
anddi
sti
l
ledwat
er
Pr
ocedur
e:
1.Takef
ourt
estt
ubesandl
abel
them as1,
2,3&4
2.Add2ml
of1%st
archsol
uti
ont
oat
estt
ube1
3.Add2ml
ofgl
ucosesol
uti
ont
otestt
ube2
4.Add2ml
ofsucr
oset
oat
hir
dtestt
ube
5.Add2ml
ofdi
sti
l
ledwat
er(
asacont
rol
)tot
hef
our
tht
estt
ube
6.Addaf
ewdr
opsofi
odi
neorpot
assi
um i
odi
desol
uti
ont
oeacht
estt
ube
7.Mi
xthecont
ent
sbyshaki
ng

Quest
ions:
1.Whatcol
ordi
dyouobser
veatt
heendofexper
iment
?
2.I
sther
eanydi
ff
erencebet
weent
het
wot
estt
ubes?

I
I.Test
sforPr
otei
ns
Act
ivi
ty1:Xant
hopr
otei
ctest
Thexant
hopr
otei
ctesti
samet
hodt
hatcanbeusedt
odet
ermi
net
heamountof
pr
otei
nsol
ubl
einasol
uti
on,usi
ngconcent
rat
edni
tr
icaci
d.Thet
estgi
vesaposi
ti
ve
r
esul
tint
hosepr
otei
nswi
thami
noaci
dscar
ryi
ngar
omat
icgr
oups,especi
all
yint
he
pr
esenceoft
yrosi
ne.I
fthet
esti
sposi
ti
vet
hepr
oofi
sneut
ral
i
zedwi
thanal
kal
i
,tur
ning
dar
kyel
l
ow.They
ell
ow col
ouri
sduet
oXant
hopr
otei
caci
dwhi
chi
sfor
medduet
o
ni
tr
ati
onofcer
tai
nami
noaci
ds,
mostcommonexampl
esbei
ngt
yrosi
neandt
rypt
ophan.
Thi
schemi
calr
eact
ioni
saqual
i
tat
ivet
est
,det
ermi
ningt
hepr
esenceorabsenceof
pr
otei
ns.Theami
noaci
dty
rosi
ne,pheny
lani
neandt
osomeext
entt
rypt
ophanear
e
r
esponsi
blef
ort
hexant
hopr
otei
ctest
.

Mat
eri
als:Testt
ubes,
testt
uber
ack,
Bunsenbur
ner
Chemi
cal
s:Eggal
bumen/
eggwhi
te,
ammoni
asol
uti
on,
nit
ri
caci
d,di
sti
l
ledwat
er

50
Pr
ocedur
e:
1.Add2ml
ofpr
otei
nsol
uti
on(
eggal
bumen)i
nat
estt
ube.
2.Addabout2ml
ofconcent
rat
edni
tr
icaci
dtot
heeggal
bumen.Mi
xcar
eful
l
y.
3.Heatt
hecont
entcar
eful
l
y.
4.Coolt
het
estt
ubeundert
apwat
erandt
hencar
eful
l
yadd2-
5mlofammoni
a
sol
uti
on(
15N)
a.Whatcol
ordi
dyounot
ewheny
oumi
xtheeggal
bumenwi
thconcent
rat
ed
ni
tr
icaci
d?
b.Di
dyouseeanycol
orchangewheny
ouheatt
hesol
uti
on?
c.Whatcol
ordi
dyou f
inal
l
ygetaf
tery
ou cool
ed t
he t
estt
ube and add
ammoni
asol
uti
ont
oit
?
Act
ivi
ty2:Bi
uretTest
Thebi
urett
esti
sachemi
calt
estusedf
ordet
ect
ingt
hepr
esenceofpept
idebonds.I
n
t
hepr
esenceofpept
ides,
acopper(
II
)ionf
ormsv
iol
et-
col
oredcoor
dinat
ioncompl
exes
i
nanal
kal
i
nesol
uti
on.Sev
eralv
ari
ant
sont
het
esthav
ebeendev
eloped,suchast
he
Modi
fi
edLowr
ytest
.Al
thoughnotv
eryspeci
fi
c,i
tisagoodgener
alt
estf
orqual
i
tat
ive
det
ermi
nat
ionofpr
otei
n.

Mat
eri
als:Testt
ubes,
testt
uber
ack,
Bunsenbur
ner
,testt
ubehol
der
Chemi
cal
s:Eggal
bumen/
eggwhi
te,
5%ofKOH/
5%ofNaOH,
dist
il
ledwat
er,
1%of
CuSO4 sol
uti
onorBenedi
ct’
ssol
uti
on

Pr
ocedur
e:
1.Add2ml
ofpr
otei
nsol
uti
on(
eggal
bumen)t
oat
estt
ube.
2.I
nanot
hert
estt
ube,
add2ml
ofdi
sti
l
ledwat
er(
asacont
rol
).
3.Addanequal
vol
umeof5%NaOHsol
uti
onandmi
x.
4.Add2dr
opsof1%CuSO4 sol
uti
onorBenedi
ct’
ssol
uti
onandmi
x.Noheat
ingi
s
r
equi
red
5.Leav
ethecont
entf
or15-
20mi
nut
es.
a.I
sther
eanycol
ordi
ff
erencebet
weent
hecont
rol
andt
hepr
otei
nsol
uti
on?
b.Whatcol
ordi
dyouobser
vei
nthepr
otei
nsol
uti
on?Howdi
dthecol
orf
orm?

51
Act
ivi
ty3:Mi
ll
on’
sTest

Mat
eri
als:Testt
ubes,
testt
uber
ack,
Bunsenbur
ner
,vor
tex,
testt
ubehol
der
Chemi
cal
s:Eggal
bumen/
ami
noaci
d,t
apwat
er,
Mil
l
on’
sreagent
Pr
ocedur
e:
1.To1ml
oft
heeggal
bumensol
uti
oni
nat
estt
ube,
addf
ewdr
opsofMi
l
lon’
s
r
eagentandusev
ort
exshaker

2.Boi
lthecont
ent
sov
eraBunsenf
lamef
or3–5mi
nut
es.

3.Cool
thecont
ent
sunderr
unni
ngt
apwat
erandaddf
ewdr
opsofsodi
um ni
tr
it
e
sol
uti
on
a. I
sther
ecol
ordi
ff
erencebet
weent
hecont
rol
andt
hepr
otei
nsol
uti
on?
b.Whatcol
ordi
dyouobser
vei
nthepr
otei
nsol
uti
on?Howdi
dthecol
orf
orm?

I
I. Te
I st
sforLi
pids

Li
pidsar
edi
ver
segr
oupofor
gani
ccompoundsi
ncl
udi
ngf
ats,oi
l
s,hor
mones,and
cer
tai
ncomponent
sofmembr
anest
hatar
egr
oupedt
oget
herbecauset
heydonot
i
nter
actappr
eci
abl
ywi
th wat
er.Mol
ecul
essuch aspr
otei
ns,nucl
eicaci
ds,and
car
bohy
drat
eshav
eanaf
fi
nit
yforwat
erandar
ecal
l
edhy
drophi
l
ic(
“wat
er-
lov
ing”
).
Li
pids,
howev
er,
arenothy
drophi
l
icbuthy
drophobi
c(“
wat
er-
fear
ing”
).
NB.Anyheat
ingi
nthathast
obedonei
nthef
oll
owi
ngt
est
sshoul
dbecar
ri
edouti
na
wat
erbat
hatt
heboi
l
ingofwat
er.Di
rectheat
ingoft
estt
ubesshoul
dnott
akepl
ace
si
ncey
oumaybr
eakt
het
estt
ube.

Act
ivi
ty1:SudanI
IIt
est
Mat
eri
als:Testt
ube,
testt
uber
ack,
testt
ubehol
der
Chemi
cal
s:SudanI
IIr
eagent
,oi
l
,wat
er

Pr
ocedur
e:
1.Add2ml
oil
to2ml
wat
eri
nat
estt
ube.
2.Addaf
ewdr
opsofSudanI
IIandshake.

52
 a.Whi
chpar
t(uporbot
tom)oft
hecont
ent
soft
het
estt
ubei
sst
ainedr
ed?
b. Whyi
sthatso?
Act
ivi
ty2:Emul
siont
est
Mat
eri
als:Testt
ube,
testt
uber
ack,
testt
ubehol
der
Chemi
cal
s:Et
hanol
,col
dwat
er,
fatoroi
l
Pr
ocedur
e:
1.Add2ml
off
atoroi
ltoat
estt
ubecont
aini
ng2ml
ofabsol
uteet
hanol
.
2.Di
ssol
vet
hel
i
pidbyshaki
ngv
igor
ousl
y.
3.Addanequal
vol
umeofcol
dwat
er.
a.Whatchangesdi
dyouobser
ve?
b.Whatef
fectdoest
heal
cohol
hav
e?

Act
ivi
ty3:Gr
easespott
est

Mat
eri
als:Api
eceofpaper
,Bunsenbur
ner
Chemi
cal
s:Et
hanol
,col
dwat
er,
fatoroi
l
Pr
ocedur
e:
1.Rubdr
opsofoi
lorf
atont
oapi
eceofpaper
.
2.Al
l
owt
imef
oranywat
ert
oev
apor
ate.Gent
lywar
mingwi
l
lspeedupt
hepr
ocess.
a.Whatdi
dyouobser
ve?
b.Whydi
dthathappen?

53
 LABORATORYSTUDYNUMBER6

BI
OCATALYSI
S–ENZYMES

6.
1Int
roduct
ion

Cat
aly
stsar
ethosesubst
ancest
hatal
tert
hespeed ofchemi
calr
eact
ion wi
thout
t
hemsel
vesunder
goi
ngper
manentchange.Fori
nst
ance,
atr
oom t
emper
atur
ehy
drogen
per
oxi
de(
H2O2)under
goesaspont
aneousbutv
erysl
ow decomposi
ti
on.Fur
ther
mor
e,
t
hisexampl
eshowst
hatnei
therpl
ati
num norcat
alasei
nit
iat
est
her
eact
ion.I
nfact
,
whatenzy
mes orcat
aly
sts do i
sto accel
erat
ethe r
ate ofanot
her
,ot
her
wise
i
mmeasur
abl
ysl
owr
eact
ion.Hy
drogenper
oxi
de(
H2O2)i
spr
oducedi
ncel
lasar
esul
tof
t
heoxi
dat
ionofmol
ecul
aroxy
geni
noxi
dat
ionr
eact
ion.Si
ncei
tist
oxi
ctocel
l
siti
s
i
mmedi
atel
ydecomposedi
ntowat
erandoxy
gen.
2H2O2→ 2H2O+O2

I
ncel
l
sthedecomposi
ti
onofhy
drogenper
oxi
dei
scat
aly
zedbyanenzy
mecal
l
ed

54
cat
alase.Thi
senzy
mei
sfoundi
nvar
ioust
issues(
E.g.Pot
ato,
li
ver
,ki
dney
,muscl
es)

Obj
ect
ives:
Af
tercompl
eti
ngt
heact
ivi
ti
est
rai
neeswi
l
lbet
o:
 ex
plai
ntheef
fectoft
emper
atur
eonenzy
meact
ivi
ty
 ex
plai
ntheef
fectofpHonenzy
meact
ivi
ty
Act
ivi
ty1:Act
ionofcat
alaseonhy
drogenper
oxi
de(
H2O2)
Mat
eri
als:Testt
ube,
pot
atocubes(
boi
l
edandunboi
l
ed)
Chemi
cal
s:3%H2O2sol
uti
on,
dist
il
ledwat
er.
Pr
ocedur
e:
1. Pl
aceabout3mlof3%H2O2sol
uti
oni
nonet
estt
ubeandanequalamountofwat
er
i
nanot
hert
estt
ube.
a.Toeachoft
het
estt
ubeaddasmal
lcubeofpot
atot
issue
b.I
nwhi
choft
het
ubei
sar
eact
ion(
gasbubbl
efor
ming)t
aki
ngpl
ace?
c.Whati
sther
eact
ionandwhati
sthepr
oduct
?
d.Whati
sther
eint
hepot
atocubet
hati
sresponsi
blef
ort
her
eact
ion?

Act
ivi
ty2:Ef
fectofhi
ght
emper
atur
eonenzy
meact
ion
Mat
eri
als:Testt
ube,
unboi
l
edpot
atot
issue,
boi
l
edpot
atot
issue.
Chemi
cal
s:H2O2
Pr
ocedur
e:
1.Taket
wocl
eant
estt
ubesandpl
aceabout3ml
of3%H2O2sol
uti
oni
neachoft
hem.
a.Pl
aceacubeofr
aw/
unboi
l
edpot
atot
issuei
nonet
estt
ubeandpl
aceacubeof
boi
l
edpot
atot
issuei
ntheot
hert
ube.
b.I
nwhi
choft
het
ubesi
sar
eact
iont
aki
ngpl
ace?
c.Howdoy
ouaccountf
ort
hef
actt
hatar
eact
ioni
staki
ngpl
acei
nonebutnoti
n
t
heot
hert
estt
ube?

Act
ivi
ty3:Ef
fectofpHonEnzy
meAct
ion
Mat
eri
als:Testt
ube,
pot
atot
issue(
boi
l
edandunboi
l
ed)

55
Chemi
cal
s:H2O2,
0.1NHCl
,0.
1NNaOH,
wat
er
Pr
ocedur
e:
1.Taket
hreecl
eant
estt
ubeandpl
ace3ml
of3%H2O2sol
uti
oni
neacht
estt
ube.
2.Label
them asA,
BandC.
3.Addt
wodr
opsof0.
1NHCli
ntestt
ubeA;
2dr
opsofwat
eri
nB;
and2dr
opsof0.
1N
NaOH i
nC.
4.Pr
epar
ethr
eepot
atocubesofequal
sizeandaddonecubet
oeachoft
het
estt
ube.
a.I
nwhi
cht
estt
ubei
sther
ateoft
her
eact
ionhi
ghest
?
b.I
nwhi
chpHr
ange(
aci
dic,
neut
ral
,oral
kal
i
ne)i
stheact
ivi
tyofcat
alasehi
ghest
?

LABORATORYSTUDYNUMBER7

PHOTOSYNTHESI
S

7.
1Int
roduct
ion
Phot
osy
nthesi
sist
he pr
ocess bywhi
ch pl
ant
s,al
gae,some bact
eri
a,and ot
her
or
gani
smsconv
ertl
i
ghtener
gyandcar
bondi
oxi
dei
ntooxy
genandgl
ucose.I
tisan
essent
ialpar
toft
hel
i
fecy
cle,andt
her
easont
hatpl
ant
sar
eatt
hebot
tom oft
hef
ood
chai
n.Ty
pical
l
y,t
hepl
ant
stakeener
gyf
rom t
hesunt
oconv
ertt
hecar
bondi
oxi
de
mol
ecul
es(
car
bonandt
wooxy
genat
oms)i
ntogl
ucosemol
ecul
es,whi
char
emadeup
ofcar
bon,hy
drogenandoxy
gen.Oxy
gengasi
sgi
venof
faswast
edur
ingt
hepr
ocess,
andwat
eri
sof
tenusedi
nthepr
ocessaswel
l
.

56
Fi
gur
e7.
1.Di
agr
ammat
icPr
ocessofPhot
osy
nthesi
s

Phot
osy
nthesi
sisdi
vi
dedi
ntot
woset
sofr
eact
ions.Somechar
act
eri
sti
csoft
hese
r
eact
ionsar
ecompar
edasf
oll
ows:

Phot
ochemi
cal
(li
ght
)react
ions Bi
ochemi
cal
(dar
k)r
eact
ions
 Li
ght
-dependent Li
ght
-i
ndependent
 Fast(
pract
ical
l
yinst
ant
aneous) Sl
ower
,butst
il
lext
remel
yfast
 Spl
i
twat
ert
orel
easeO2, Conv
ert(
fi
x)CO2t
osugar
El
ect
ronsandpr
otons

57
Fi
gur
e7.
2.Di
agr
ammat
icpr
esent
ati
onofpl
antpi
gment
ati
ons

Object
ives:
Uponcompl et
ionoft
heseact
ivi
ti
es,
youar
eexpect
edt
o:

58
  i
sol
ateandi
dent
if
yphot
osy
nthet
icpi
gment
sinl
eav
es
 cal
cul
ater
etent
ionf
act
or(
Rf)v
aluesofphot
osy
nthet
icpi
gment
sandgr
apht
he
absor
pti
onspect
rum f
oreachpi
gment
 i
nvest
igat
ethei
mpor
tanceofl
i
ghtandchl
orophy
llf
orst
archpr
oduct
ion

Act
ivi
ty1:Paperchr
omat
ogr
aphyofl
eafpi
gment

Materi
als:
.Greenleaves,
chromatographychamber,
mortarandpestl
e,sci
ssor
s,penci
l
,
capi
ll
ary
tube,
spatula,
scale,
fil
terpaperst
ri
ps,
watchglasses,
thr
ead

Chemi
cal
s:Et
hanol
,pet
rol
eum et
her
/acet
one

Pr
ocedur
e:

1.Pr
epar
eleafext
ract

2.Cutoutast
ri
poff
il
terpaper
/chr
omat
ogr
aphyst
ri
pabout2cm wi
de

3.Dr
awahor
izont
all
i
newi
thapenci
labout2-
3cm awayf
rom t
het
ipoft
henot
ch

4.Usi
ngacapi
l
lar
ytube,
placeaf
ewdr
opsoft
heext
ractont
othemi
ddl
eoft
hel
i
ne
andl
eti
tdr
y.

Not
e:eachdr
opshoul
dbeal
l
owedt
odr
ybef
orepl
aci
ngt
henextone.

5.Add2ml
ofchr
omat
ogr
aphysol
vent(
9par
tspet
rol
eum et
her
:1par
tacet
one)

6.I
mmer
set
heendoft
hest
ri
pint
hechr
omat
ogr
aphi
cchamberj
ustabov
ethe
l
evel
oft
hesol
ventandwat
chast
hesol
ventmov
esupt
hepaper
.

Thespotshoul
dnotbeundert
hesur
faceoft
hesol
vent
.

7.Remov
echr
omat
ogr
aphyst
ri
pbef
oret
hesol
vent
-f
rontr
eachest
het
opoft
he
st
ri
p(af
ter3t
o5mi
nut
es)
.Mar
kwi
thapenci
lthedi
stancet
hatt
hesol
ventf
ront
t
rav
ell
ed,
andsett
hest
ri
pasi
dedr
y.Youwi
l
lseef
ourbandsofcol
ouront
hest
ri
p:
ay
ell
owbandofcar
otenesneart
het
op,
fol
l
owedbyay
ell
owbandof
xant
hophy
ll
ous,
abl
ue-
greenbandofchl
orophy
lla,
anday
ell
ow-
greenbandof

59
 chl
orophy
llbneart
hebot
tom.

Quest
ions

1.Measur
ethedi
stanceofeachpi
gmentbandf
rom t
hel
oadi
ngspotandal
so
t
hedi
stancet
rav
ell
edbyt
hesol
vent
.Cal
cul
atet
heRfv
alueusi
ngt
heequat
ion
andr
ecor
dthev
aluesi
nthet
abl
e.

Not
e:AnRfv
aluet
hati
scl
oset
o1i
ndi
cat
est
hatt
hepi
gmenti
sver
ysol
ubl
eint
he
sol
vent
.AnRfv
aluet
hati
sver
ysmal
lindi
cat
est
hatt
hepi
gmenti
snotv
erysol
ubl
ein
t
hesol
vent
.

RFv alues Pigmentidenti

fi
cat i
on
β-carotene=0.99 Chlorophyl
la=blue-green
Chlorophy l
la=0.30 Chl
or ophy
llb=Olivegr een
Chlorophy l
lb=0.13 Xanthophyl
l=yell
ow
Violaxanthin=0.
40 Carotene=orangeyellow
Lutein=0. 68

2.Whatdoesasmal
lRfv
aluet
ell
youaboutt
hechar
act
eri
sti
csoft
hemov
ingmol
ecul
es?

3.Whyi
schr
omat
ogr
aphyusef
ul?

Fact
orsnecessar
yforphot
osy
nthesi
s

Chl
orophy
ll
,li
ght
,Car
bondi
oxi
de,wat
er,andt
emper
atur
ear
emaj
orf
act
orsi
mpor
tant
f
orphot
osy
nthesi
stot
akepl
ace.
Act
ivi
ty1:Li
ght–af
act
ornecessar
yforphot
osy
nthesi
s

60
Mat
eri
als:Par
tl
ymaskedl
eaf
,Pet
ri
-di
sh
Chemi
cal
s:wat
er,
iodi
nesol
uti
on,
alcohol

Pr
ocedur
e:

1.Youwi
l
lbepr
ovi
dedwi
thal
eafwhi
ch,
whi
l
est
il
lat
tachedt
oani
ntactpl
anti
spar
tl
y
maskedf
ort
hel
ast24–48hour
spar
tl
ycl
i
ppedandcov
ered.
2. Remov
ethecov
erandcar
ryoutt
het
estf
orst
archasdescr
ibedbel
ow.
3.Keept
hel
eafi
nboi
l
ingwat
erf
orabout5mi
nut
es.Thi
ski
l
lst
hecel
l
sandr
ender
s
t
hem hi
ghl
yper
meabl
e.
4.Tr
ansf
ert
hel
eafi
nwar
m al
cohol
;andkeepi
tther
eunt
ili
tisdecol
ori
zed.The
al
coholi
skeptwar
minawat
erbat
h,ori
tmaybedi
rect
lywar
medov
era‘
cont
rol
l
ed’
ov
enorhot
-pl
ate.
5.Soakt
hedecol
ori
zedl
eafi
nwat
er.Thenputi
taPet
ridi
shandcov
eri
twi
th0.
05
i
odi
nesol
uti
on.
6. Al
l
owt
hel
eaft
oremai
nint
hei
odi
nesol
uti
onf
or5mi
nut
esandt
henr
inse(
wash)i
t
wi
thwat
er.

Quest
ions:

1.Whi
chpar
toft
hel
eaf
,themaskedort
heunmasked,
isposi
ti
vef
orst
arch?
2.Whatdoest
hisr
esul
tshowaboutt
hei
mpor
tanceofl
i
ghtf
orst
archpr
oduct
ion?

Act
ivi
ty2:Chl
orophy
ll-af
act
ornecessar
yforphot
osy
nthesi
s

Mat
eri
als:Var
iegat
edl
eaf
,Pet
ridi
sh

Chemi
cal
s:wat
er,
iodi
nesol
uti
on,
alcohol

Pr
ocedur
e:

1.Youwil
lbeprovi
dedwi
thav
ari
egat
edl
eaf
.Makeadr
awi
ngoft
hedi
str
ibut
ionof
col
orsi
n thel
eaf.
2.Testt
hel
eaff
orst
arch.
3.Makethedr
awi
ngoft
hedi
str
ibut
ionofst
archi
nthel
eafandcompar
eitwi
thf
ir
st
drawi
ng.
Quest
ions:
61
1.Howdot
hedi
str
ibut
ionofchl
orophy
llandofst
archi
nthel
eafar
ecompar
ed?

2.I
schl
orophy
lli
mpor
tantf
orphot
osy
nthesi
s?Expl
aint
hisi
nli
newi
thy
ourr
esul
t.

LABORATORYSTUDYNUMBER8

MI
CROBI
OLOGI
CALLABORATORYTECHNI
QUES

8.
1Int
roduct
ion
I
nthel
abor
ator
y,t
henut
ri
entpr
epar
ati
onst
hatar
eusedf
orcul
tur
ingmi
croor
gani
sms
ar
ecal
l
edmedi
a(si
ngul
ar,medi
um)
.Thr
eephy
sicalf
ormsar
eused:Li
qui
d(br
oth)
medi
a,semi
‐sol
i
dmedi
aandsol
i
dmedi
a.Themaj
ordi
ff
erenceamongt
hesemedi
ais
t
hatsol
i
dandsemi
‐sol
i
d cont
ainasol
i
dif
ying agentcal
l
edagar
,wher
easal
i
qui
d
medi .Agari
um doesnot spol
ysacchar
ideext
ract
edf
rom mar
ineseaweedssuchasr
ed
al
gae.Agarhassomeuni
quepr
oper
ti
est
hatmakei
tusef
uli
ncul
tur
emedi
a.Onl
yfew
mi
crobescandegr
adeagarsoi
tremai
nssol
i
ddur
ingmi
crobi
algr
owt
h.I
tli
quef
iesat
100oCandr
emai
nsi
nal
i
qui
dst
ateunt
ilcool o40oC.1.
edt 5%agarpr
oducesaf
ir
m gel
39oCbut
at32‐ ,oncesol
i
dif
ied,
itwi
l
lnotmel
tunt
il85oCi
sreached.Thi
smeansy
oucan
i
ncubat
epl
atesatanyt
emper
atur o85oC.Youcanaddor
eupt gani
smst
omol
tenagar

62
cool
edt
oat
emper
atur
e( 45oC)whi
40‐ chwi
l
lnotdamaget
hemaj
ori
tyofor
gani
sms.
Thi
sal
l
owst
hepourpl
atet
echni
quet
obecar
ri
edout
.

Whi
l
eint
hel
i
quef
iedst
ate,sol
i
dmedi
acanbepour
edi
ntoei
therat
estt
ubeorPet
ri
pl
ate.I
fthemedi
um i
sal
l
owedt
ohar
deni
nasl
ant
edposi
ti
on,
thet
ubei
sdesi
gnat
edas
Agarsl
ant
.Ift
het
ubei
sal
l
owedt
ohar
deni
nanupr
ightposi
ti
on,
thet
ubei
sdesi
gnat
ed
as Agardeep t
ubeand i
ftheagari
spour
ed i
nto a Pet
ri
‐pl
ate,t
hen t
hepat
eis
desi edanAgarpl
gnat ate.
Mostchemo-
het
erot
rophi
cbact
eri
aar
erout
inel
ygr
ownoncompl
exmedi
a.Or
gani
c
car
bon,ener
gy,ni
tr
ogensour
cesar
eusual
l
ysuppl
i
edbypr
otei
nint
hef
orm ofmeat
ext
ract
sandpar
ti
all
ydi
gest
edpr
otei edpept
nscal
l ones.

The pr
epar
ati
on ofmedi
afr
om commer
cialdehy
drat
ed pr
oduct
sis si
mpl
e and
st
rai
ght
for
war
d.Eachbot
tl
eofdehy
drat
edmedi
um hasi
nst
ruct
ionsf
orpr
epar
ati
onon
i
tsl
abel
.Medi
amustbest
eri
l
izedaf
terpr
epar
ati
on.Themostcommonmet
hodof
st
eri
l
izi
ngcul
tur
emedi
athatar
eheatst
abl
eisst
eam st
eri
l
izat
ion,
oraut
ocl
avi
ng,
usi
ng
st
eam underpr
essur
e.Dur
ingt
hispr
ocess,mat
eri
alt
obest
eri
l
izedi
spl
acedi
nthe
aut
ocl
aveandheat o121oCat15poundsofpr
edt essur
efor15mi
nut
es.

Obj
ect
ives:
Af
tercompl
eti
ngt
heact
ivi
tyst
udent
swi
l
lbeabl
eto:
 Pr
epar
edi
ff
erentcul
tur
emedi
a
 descr
ibet
hei
mpor
tanceofst
eri
l
izat
ioni
nmi
crobi
alt
echni
ques
 i
sol
atebact
eri
ausi
ngdi
ff
erentt
echni
quest
ogetpur
ecul
tur
e
 pr
epar
eabact
eri
alsmearpr
oper
ly
 st
ainbact
eri
aandobser
vet
hem undert
hemi
croscope

Act
ivi
ty1:Pr
epar
ati
onofcul
tur
emedi
um
Mat
eri
als:Pet
ripl
ate,aut
ocl
ave,er
lenmey
erf
lask,t
ri
ple beam bal
ance,i
ncubat
or,
al
umi
num f
oil
,testt
ubes,
hotpl
ate,
magnet
icst
ir
rer
,heatpr
oofgl
oves,
saf
ety
hood/
saf
etycabi
net
,cot
ton,spr
itl
amp,met
ali
nocul
ati
ngl
oop,per
manent

63
 mar
ker
,l-
shapedgl
assr
od,v
olumet
ri
cpi
pet
tes,mi
cropi
pet
te,pi
pet
tef
il
ler
,
spr
eadi
ngwheel
,cent
ri
fuge,
bot
tl
es(
reagentbot
tl
es)
Chemi
cal
s:Nut
ri
entAgar
,Nut
ri
entBr
oth,
Pot
atoDext
roseAgar
,Al
cohol

Pr
ocedur
e

1.Wei
gh7gr
amsoft
hepowderoft
heNut
ri
entagargi
ven(
asment
ionedont
he
l
abel
oft
hebot
tl
e)andaddi
tto500ml
flask.
2.Add250ml
ofdi
sti
l
ledwat
er
3.UseAl
umi
num f
oil
orcot
tont
ocov
ert
hemout
hoft
hef
lask.
4.Heatt
hesol
uti
onunt
ili
tbegi
nst
oboi
l(Makesur
etousemagnet
icst
ir
rerort
o
shaket
hebot
tl
eper
iodi
cal
l
yunt
ili
tboi
l
s)
5.Loadtheautocl
avewi
tht
hepr
epar
edcul
tur
emedi
aandaut
ocl
avef
or15mi
nut
es
o
ormoreat121C
6.Whent
heper
iodofst
eri
l
izat
ioni
scompl
etedandt
hepr
essur
eint
hechamber
r
eads0,
car
eful
l
yopent
hedoorandr
emov
ethecont
ainer
s.
Bef
oreopeni
ngt
heaut
ocl
ave,
youshoul
dwearheat
‐pr
oofgl
oves,
standatar
m’s
l
engt
h,andsl
owl
yopent
hedoor
.
7.Wai
tunt
ilt
het
emper
atur
edr
opsappr
oxi
mat
elyt 50oC
o48‐
Pour15‐20mlofthemedium t
osteri
le,
label(
indi
catingt
henameofthemedium)Petri
‐
pl
atesinaLaminarfl
owfoll
owingasepti
cprocedures.Waitunt
ilt
heagarsol
i
dif
iesand
i
nvertt
heplat
esupsidedown.

Quest
ions:

1. Whyar
ecul
tur
emedi
ast
eri
l
izedbef
oreuse?
2. Whyi
sagarPet
ri
‐pl
atesi
nver
tedaf
tert
heyhav
ecool
ed?
3. Whyi
scul
tur
emedi
um cool oabout48oCt
edt o50oCbef
orepour
ing?
4. Whatar
ethet
hreemai
nty
pes(
int
ermsoft
hei
rphy
sicalf
orms)ofmi
crobi
ologi
cal
cul
tur
e medi
a?

Act
ivi
ty2:I
sol
ati
ont
echni
quesofpur
ebact
eri
alcul
tur
e

A.St
reakpl
atet
echni
que
I
nnat
uremostmi
crobesar
efoundgr
owi
ngi
nenv
ironment
sthatcont
ainmanydi
ff
erent

64
or
gani
sms.Howev
er,mi
xed cul
tur
es ar
e ofl
i
ttl
e use i
n st
udy
ing mi
croor
gani
sms
becauseoft
hedi
ff
icul
tyt
heypr
esenti
ndet
ermi
ningwhi
chor
gani
sm i
sresponsi
blef
or
anyobser
ved act
ivi
ty.A pur
e cul
tur
eisa cul
tur
ethatcont
ainsonl
yone ki
nd of
or
gani
sm.A pur
e cul
tur
eisr
equi
red i
n or
dert
o st
udyconcept
s such as gr
owt
h
char
act
eri
sti
cs,pat
hogeni
cit
y,met
abol
i
sm,and ant
ibi
oti
c suscept
ibi
l
ity and any
bi
ochemi
calt
est
s.Ther
ear
ethr
eedi
l
uti
onmet
hodscommonl
yusedf
ort
hei
sol
ati
onof
bact
eri
a:t
hest
reakpl
ate,
thespr
eadpl
ate,
andt
hepourpl
ate.
I
nthest
reakpl
atet
echni
que,al
oopi
susedt
ost
reakt
hemi
xedsampl
emanyt
imes
ov
ert
hesur
faceofasol
i
dcul
tur
emedi
um i
naPet
ri
‐pl
ate.Atsomepoi
ntont
hest
reaks,
i
ndi
vi
dualcel
l
swi
l
lber
emov
edf
rom t
hel
oopasi
tgl
i
desal
ongt
heagarsur
faceandwi
l
l
gi
ver
iset
osepar
atecol
oni
es.Oneassumest
hatonecol
onycomesf
rom onecel
l
.The
pr
inci
plei
sthatbyst
reaki
ng,adi
l
uti
ongr
adi
enti
sest
abl
i
shedont
hesur
faceoft
he
pl
ateascel
l
sar
edeposi
tedont
heagarsur
face.St
reakpl
atei
sthemostcommon
i
sol
ati
ont
echni
quei
nuset
oday
.
Mat
eri
als:Spr
itl
amp,
inocul
ati
ngl
oops,
per
manentmar
ker
Chemi
cal
s:24‐
48hour
smi
xednut
ri
entbr
othcul
tur
es,
nut
ri
entagarpl
ates
Pr
ocedur
e:
1.Labelal
lthepl
ates,ont
hebot
tom,wi
tht
henameoft
heor
gani
sm(
s)orsampl
e
codet
obei
nocul
ated,
thedat
eandy
ournamewi
thmar
ker
.
2.Fl
amet
hei
nocul
ati
ngl
oopt
oredness,al
l
ow i
ttocool
,andasept
ical
l
yobt
aina
l
oopf
ulofonebr
othcul
tur
e.
3.Tost
reakapl
ate,
li
ftoneedgeoft
hePet
ripl
atecov
er,
andst
reakt
hef
ir
stsect
or
bymaki
ngasmanyst
reaksaspossi
blewi
thoutov
erl
appi
ngpr
evi
ousst
reaks.Do
notgouget
heagarwhi
l
est
reaki
ngt
hepl
ate.
4.Fl
amey
ourl
oopandl
eti
tcool
.Tur
nthepl
atesot
henextsect
ori
sont
op.St
reak
t
hroughonear
eaoft
hef
ir
stsect
or,andst
reakaf
ewt
imesawayf
rom t
hef
ir
st
sect
or.
5.Fl
amey
ourl
oop,t
urnt
hepl
ateagai
n,andst
reakt
hroughonear
eoft
hesecond
sect
or.Thenst
reakt
het
hir
dsect
or.
6.Fl
amey
ourl
oop,st
reakt
hroughonear
eaoft
het
hir
dsect
or,andt
henst
reakt
he
r
emai
ningar
eaoft
heagarsur
face,bei
ngcar
efulnott
omakeaddi
ti
onalcont
act

65
 wi
thanyst
reaksi
nthepr
evi
oussect
ions.Fl
amey
ourl
oopbef
oreset
ti
ngi
tdown.
(
Why
?)
7.I
ncubat
et heplatesi
naninvert
edposi
ti
oninthe35oC i
ncubat
or(
oratr
oom
t
emperat
ure,dependi
ngont
heinocul
um)f
or24‐
48hours.

Fi
gur
e8.
1:Thest
reaki
ngt
echni
quef
orpur
eisol
ati
onofbact
eri
alcul
tur
e

Quest
ions:

1.Whatist
heneedtolabel
theplatesont hebot
tom?
2.Whyi si
tnecessar
yt oincubat
ei noculat
edagarpl
atecul
tur
esi
nani
nver
ted
posi
ti
on?

Act
ivi
ty3:Smearpr
epar
ati
onandsi
mpl
est
aini
ng
A.Smearpr
epar
ati
on

66
Mat
eri
als:Mi
croscope,
micr
oscopi
csl
i
des,
cov
ersl
i
ps,
inocul
ati
ngl
oopandneedl
e,
spr
itl
amp,
per
manentmar
ker
,for
ceps,
blot
ti
ngpaper
Chemi
cal
s:Agarpl
atecul
tur
e,st
eri
l
edi
sti
l
ledwat
eri
nadr
opper
,oi
li
mmer
sion,
cry
stal
v
iol
etandmet
hyl
enebl
ue
Pr
ocedur
e:

1.Mar
kthename(
Code)oft
hebact
eri
alcul
tur
eint
hef
arcor
neroneachoft
he
sl
i
des.
2.Fora br
oth cul
tur
e,shake t
he cul
tur
etube and,wi
th an i
nocul
ati
ng l
oop,
asept
ical
l
ytr
ansf
er1t
o2l
oopf
ulsofbact
eri
atot
hecent
eroft
hesl
i
de.Spr
ead
t
hisoutt
oabouthal
finch(
1.27cm)ar
ea.Whenpr
epar
ingasmearf
rom asl
ant
orpl
ate,pl
aceal
oopf
ulofwat
eri
nthecent
eroft
hesl
i
de.Wi
tht
hei
nocul
ati
ng
needl
e,asept
ical
l
ypi
ckupav
erysmal
lamountofcul
tur
eandmi
xint
othedr
op
ofwat
er.Spr
eadt
hisoutasabov
e.
3.Al
l
owt
hesl
i
dest
oai
rdr
y.
4.Passt
hesl
i
dest
hroughaBunsenbur
nerorspr
itl
ampf
lamet
hreet
imest
oheat
f
ixandki
l
lthebact
eri
a.

B.Si
mpl
est
aini
ng
Pr
ocedur
e:
1.Pl
acethefi
xedsmear
sonast
aini
ngl
ooporr
ackov
erasi
nkorot
hersui
tabl
e
r
ecept
acl
e.
2.St
aint
hesl
i
dewi
thanyoft
het
hreest
ainsgi
ven(
i.
e.,cr
yst
alv
iol
et(
20t
o30
secondsst
aini
ngt
ime)
,Car
bol
fuchsi
n(5t
o10secondsst
aini
ngt
ime)
,oral
kal
i
ne
Met
hyl
enebl
ue(
1mi
nut
est
aini
ngt
ime)
).
3.Washst
ainof
fsl
i
dewi
thwat
erf
oraf
ewseconds.
4.Bl
otsl
i
dedr
ywi
thbl
ott
ingpaper
.Becar
efulnott
orubt
hesmearwhendr
yingt
he
sl
i
debecauset
hiswi
l
lremov
ethest
ainedbact
eri
a.
5.Exami
neundert
heoi
li
mmer
sionl
ens.

Quest
ions:
1.Whatwoul
dhappeni
fyouappl
ytomuchheatwhi
l
eheatf
ixi
ngt
hesmear
?
2.Ment
iont
hreei
mpor
tanceofheatf
ixi
ng?

67
 3.Whati
stheadv
ant
ageofsi
mpl
est
aini
ngov
erwetmount
?
4.Whati
sthebi
ochemi
cal
basi
sforsi
mpl
est
aini
ng?
C.Gr
am’
sst
aini
ng
Bact
eri
adi
ff
erf
rom oneanot
herchemi
cal
l
yandphy
sical
l
yandmayr
eactdi
ff
erent
lyt
oa
gi
venst
aini
ngpr
ocedur
e.Thi
sist
hebasi
sfordi
ff
erent
ialst
aini
ng.Gr
am st
aini
sa
di
ff
erent
ialst
ainasi
tdi
ff
erent
iat
esbact
eri
aint
wogr
oups‐gr
am negat
iveandgr
am
posi
ti
ve.TheGr
am st
ain(
named af
terHansChr
ist
ianGr
am,Dani
shSci
ent
istand
phy
sici
an)i
sthemostusef
ulandwi
del
yempl
oyeddi
ff
erent
ialst
aini
nbact
eri
ology
.
Gr
am st
aini
ngi
sthesi
ngl
emostusef
ult
esti
nthecl
i
nical
micr
obi
ologyl
abor
ator
y.

Thef
ir
stst
epi
nthepr
ocedur
einv
olv
esst
aini
ngwi
tht
hebasi
cdy
ecr
yst
alv
iol
et.Thi
sis
t
hepr
imar
yst
ain.I
tisf
oll
owedbyt
reat
mentwi
thani
odi
nesol
uti
on,
whi
chf
unct
ionsas
amor
dant
;thati
s,i
tincr
easest
hei
nter
act
ionbet
weent
hebact
eri
alcel
landt
hedy
eso
t
hatt
hedy
eismor
eti
ght
lyboundort
hecel
li
smor
est
rongl
yst
ained.Thesmeari
sthen
decol
ori
zedbywashi
ngwi
th95% et
hanol
.Gr
am posi
ti
vebact
eri
aret
aint
hecr
yst
al
v
iol
et‐i
odi
necompl
exwhen washed wi
tht
hedecol
ori
zer
,wher
easgr
am–negat
ive
bact
eri
aloset
hei
rcr
yst
alv
iol
et–i
odi
necompl
exandbecomecol
orl
ess.Fi
nal
l
y,t
he
smeari
scount
erst
ainedwi
thabasi
cdy
e,di
ff
erenti
ncol
orf
rom cr
yst
alv
iol
et.Thi
s
count
erst
aini
susual
l
ysaf
rani
n.Thesaf
rani
nwi
l
lst
aint
hecol
orl
ess,gr
am negat
ive
bact
eri
api
nkbutdoesnotal
tert
hedar
kpur
plecol
oroft
hegr
am posi
ti
vebact
eri
a.

Gr
am st
aini
smor
econsi
stentwi
thy
oungcul
tur
es(
8‐16hour
sol
d).Ol
dcul
tur
esmay
show v
ari
ati
oni
nGr
am st
aini
ng.Besi
des,somebact
eri
alspeci
esar
egr
am v
ari
abl
e.
Thati
s,somecel
l
sint
hesamecul
tur
ewi
l
lbegr
am posi
ti
veandsome,gr
am negat
ive.
Hence,i
ndi
sti
nctgr
am‐st
ainr
esul
tscanbeconf
ir
medbyasi
mpl
etestusi
ngKOH.
Pl
aceadr
opof10% KOH onacl
eangl
asssl
i
deandmi
xwi
thal
oopf
ulofbact
eri
al
past
e.Wai
t30secondst
henpul
lthel
oopsl
owl
ythr
ought
hesuspensi
onupandaway
f
rom t
hesl
i
de.Agr
am negat
iveor
gani
sm wi
l
lpr
oduceamucoi
dst
ri
ngwher
easgr
am‐
posi
ti
veor
gani
sm r
emai
nsf
lui
d.

Mat
eri
als:Mi
croscope,
glasssl
i
des,
inocul
ati
ngl
oop,
spr
itl
amp,
blot
ti
ngpaper
,st
aini
ng

68
r
ack
Chemi
cal
s:Sol
uti
onsofCr
yst
alv
iol
et,Gr
am’
siodi
ne(2gKIi
n300mlDi
sti
l
ledwat
er
pl
us1g,i
odi
necr
yst
al)
,95% et
hanol
,Saf
rani
n,KOH (
3%)
,18‐
24hournut
ri
ent
br
othcul
tur
esofGr
am posi
ti
veandGr
am negat
ivebact
eri
a,i
mmer
sionoi
l
Pr
ocedur
e

1.Pr
epar
eheat
‐f
ixedsmear
soft
hegi
vencul
tur
eandmar
kci
rcl
ear
oundeach
smearont
her
ever
sesi
deofsl
i
dewi
thmar
ker
.
2.Pl
acet
hesl
i
desont
hest
aini
ngr
ack.
3.Fl
oodt
hesmear
swi
thcr
yst
alv
iol
etandl
etst
andf
or30seconds.
4.Ri
nsewi
thwat
erf
or5seconds.
5.Cov
erwi
thGr
am’
siodi
nemor
dantandl
eti
tst
andf
or1mi
nut
e.
6.Ri
nsewi
thwat
erf
or5seconds.
7.Decol
ori
zewi
th95%et
hanolf
or15‐30seconds.Donotdecol
ori
zet
ool
ong.Add
t
hedecol
ori
zerdr
opbydr
opunt
ilt
hecr
yst
alv
iol
etf
ail
stowashf
rom t
hesl
i
de.
8.Ri
nsewi
thwat
erf
or5seconds.
9.Count
erst
ainwi
thsaf
rani
nforabout60‐
80seconds.
10.
Rinsewi
thwat
erf
or5seconds
11.
Blotdr
yandexami
neunderoi
li
mmer
sion.Gr
am posi
ti
veor
gani
smsst
ainbl
uet
o
pur
ple;gr
am negat
iveor
gani
smsst
ainpi
nkt
ored.Ther
eisnoneedt
opl
acea
cov
ersl
i
pont
hest
ainedsmear
.

Quest
ions:
1.Whati
sthebasi
sfort
heGr
am st
aint
echni
que?
2.Whyi
stheGr
am st
aint
echni
queknownasadi
ff
erent
ial
techni
que?
3.Whatdoy
ouconcl
udei
fyouf
indr
edandpur
plecocci
inpur
ecul
tur
e?

69
 LABORATORYSTUDYNUMBER9

CELLDI
VISI
ON

9.
1Int
roduct
ion
Cel
ldi
vi
sioni
sapr
ocesswher
ecel
l
srepr
oduceandmul
ti
plyt
hei
rnumber
.Mi
tosi
sisa
nucl
eardi
vi
sion,whi
chpr
oducest
wocompl
eteset
sofchr
omosomes.Eachseti
slat
er
separ
atedf
rom t
heot
herbycy
toki
nesi
s.Thenewl
yfor
meddaught
ercel
l
sar
eident
ical
t
ooneanot
her
.Thi
sdi
vi
sional
socal
l
edequat
ionaldi
vi
sion,ast
hechr
omosomesar
e
equal
l
ydi
str
ibut
edamongt
het
wodaught
ercel
l
s.Thi
sdi
vi
sionensur
est
hatani
dent
ical
genet
icconst
it
uti
oni
smai
ntai
nedf
oral
lthecel
l
soft
hebody
.

Mostoft
hedi
ploi
dor
gani
smsunder
gor
educt
iondi
vi
siondur
inggamet
ogenesi
si.
e.,
t
heypr
oducegamet
est
hatar
ehapl
oid.Thecel
ldi
vi
siont
hatbr
ingsaboutt
hisr
educt
ion
f
rom t
woset
sofchr
omosomes(
dipl
oid)t
oonesetofchr
omosomes(
hapl
oid)i
scal
l
ed
mei
osi
s.Mei
osi
scompr
isesoft
wodi
vi
sions-Mei
osi
sI&Mei
osi
sII
.Att
heendof
mei
osi
sfourdaught
ercel
l
s ar
e pr
oduced,each daught
ercel
lhav
ing hal
fthe
chr
omosomecompl
ement–oft
hepar
ent
.

Obj
ect
ives:
 Af
tercompl
eti
ngt
heact
ivi
ti
est
rai
neeswi
l
lbeabl
eto
 descr
ibet
hepr
ocessofmi
tot
iccel
ldi
vi
sion
 descr
ibet
hepr
ocessofmei
oti
ccel
ldi
vi
sion
 di
scusst
hedi
ff
erencesbet
weenmi
tot
iccel
ldi
vi
sionandmei
oti
ccel
ldi
vi
sion

Act
ivi
ty1:Exper
imentonMi
tosi
s

Mat
eri
als:Mi
croscope,
sli
desandcov
ersl
i
ps,
ger
minat
ingoni
onorgar
li
c,t
estt
ubes,
t
estt
ubehol
der
s,wat
chgl
ass,
Bunsenbur
ner
Chemi
cal
s:Di
sti
l
ledwat
er,
acet
ocar
mine/
haemat
oxy
li
n,0.
1NHCl
,20%acet
icaci
d,
Cor
noy
’sf
lui
d(30ml
of100%ofEt
hyl
alcohol
,5ml
ofgl
aci
alacet
icaci
d,15ml
of
chl
orof
orm)

Pr
ocedur
e:

70
 1.Separ
atet
he gar
li
c cl
oves (
oruse oni
ons)and pl
ace t
hem i
ntestt
ubes
cont
aini
ngwat
er,
sucht
hatt
hei
rbasesar
edi
ppi
ngi
nwat
er.
2.Wi
thi
n2-
3day
sroott
ipsst
artappear
ing.
3. Cutt
her
oott
ipsandf
ixt
hem i
ncar
noy
’sf
lui
d(100%et
hylal
cohol
-30ml
;gl
aci
al
acet
icaci
d-5ml
;Chl
orof
orm-
15ml
),f
or18-
24hr
satr
oom t
emper
at eor4oC.
ur
Thent
ransf
ert
hem t
opr
eser
vat
ivesuchasspr
itorf
ormal
i
n
4.Remov
ether
oott
ipsandkeept
hem i
nwat
eri
nawat
chgl
assf
oraf
ewmi
nut
es.
0
5. Remov
ethewat
erandadd0.
1NHClandheati
tinwat
erbat
h(about60C)and
wai
tforabout10mi
nut
esunt
ilt
her
oott
ipsbecomesof
t.(
Donotal
l
ow t
he
mat
eri
alt
oboi
l
).
6. Remov
etheHClandwasht
hemat
eri
alwi
thdi
sti
l
ledwat
er,2-
3ti
mest
il
lal
la
t
raceoft
heaci
disr
emov
ed.
7.Addt
hest
ain-
acet
ocar
mine/
haemat
oxy
li
nandal
l
owt
hemat
eri
alt
oremai
nint
he
st
ainf
orabout10-
15mi
nut
es.
8.Cutt
heswol
l
en(
stai
ned)r
oott
ipsandpl
acet
hem onacl
eansl
i
de.Add2-
3dr
ops
of20%acet
icaci
d.Cov
eri
twi
thacov
ersl
i
pandt
apgent
lyt
omakeasquash.
9.Addadr
opoff
reshst
ainont
her
oott
ipandcov
eri
twi
thcov
ersl
i
p
10.Tapont
hecov
ersl
i
pgent
lywi
tht
her
ubberendofy
ourpenci
l
.Thi
swi
l
lbr
eakt
he
t
issueapar
t.
11.Pl
aceaf
oldedpi
eceofsof
tpaperont
hecov
ersl
i
pandsquasht
her
oott
ipby
appl
yi
ngauni
for
m pr
essur
eont
hecov
ersl
i
pwi
tht
hebal
lofy
ourt
humb.
I
fyouf
indy
ourpr
epar
ati
onunst
ained,
rest
aini
tusi
ngt
hef
oll
owi
ngst
eps:
12.
Placeadr
opoft
hest
ainatoneedgeoft
hecov
ersl
i
p
13.Byhol
dingapi
eceofsof
tpaperoranyabsor
bentpaperatt
heopposi
teend,
dr
awt
hest
ainundert
hecov
ersl
i
p
14.
Obser
vet
hesl
i
deundert
hemi
croscopeandi
dent
if
ythedi
ff
erentst
agesof
mi
tosi
s.
NB.Wheny
ouobser
vet
hedi
vi
dingcel
l
sundert
hemi
croscope,
taket
hef
oll
owi
ngi
nto
consi
der
ati
on:
a. Thet
reat
menthaski
l
ledt
hecel
l
s.Ther
efor
e,onecel
lshowsonl
yonephasesi
.e.
t
hephasei
nwhi
chi
thadbeenbef
oret
het
reat
ment
.

71
 b.Youmaynotf
indal
lthephasei
nanysi
ngl
efl
i
edofv
isi
on
c. Donotexpectt
ofi
ndt
hephasesi
ntheor
derI
nter
phase,Pr
ophase,Met
aphase
andTel
ophase
d. I
nanyonef
iel
dofv
isi
ont
her
ear
eal
argenumberofcel
l
s,usi
ngt
hel
owpower
ort
hemi
ddl
epower
,scant
hrought
hef
iel
dwi
thy
ourey
es.Wheny
ouf
indacel
li
n
t
hephasey
ouar
elooki
ngf
or,br
ingi
ttot
hecent
eroft
hef
iel
d.Thenshi
ftt
he
hi
ghpowerobj
ect
ive.

Quest
ions:
1.Whyy
ouusedt
heswol
l
enr
oott
ipt
oobser
vet
hest
agesofcel
ldi
vi
sion?
2.Whati
s/ar
ethest
age(
s)ofmi
tosi
sobser
ved?

Act
ivi
ty2:Exper
imenton:Mei
osi
s

Mat
eri
als:Mai
zeort
est
isofgr
asshopper
,(f
reshant
her
sfr
om unopenedbudscanal
so
beused)
,Mi
croscope,
sli
desandcov
ersl
i
ps,
Bunsenbur
ner
Chemi
cal
s:Car
noy
’sf
lui
dorf
ormal
i
n-acet
ic/
propi
oni
c-al
cohol(
90mlof50% ofet
hyl
al
cohol
),5mlofgl
aci
alacet
icaci
dor5mlofPr
opi
oni
caci
d,5mloff
ormal
i
n,
acet
ocar
mine/
haemat
oxy
li
n

Pr
ocedur
e:
1.Fr
eshorf
ixedant
her
sofmai
zeort
het
est
isofgr
asshopperi
stakenonacl
ean
sl
i
de.
2.Themat
eri
ali
scr
ushedwi
thascal
peloranot
hersl
i
desucht
hatt
hecel
l
sar
e
spr
eadorsmear
edi
nasi
ngl
elay
er.
3. I
faf
reshmat
eri
ali
sused,i
nver
tthesl
i
deov
eraPet
ridi
shcont
aini
ngt
hef
ixi
ng
andki
l
li
ngagent(
Car
noy
,sf
lui
dorFAA)
,sucht
hatt
hemat
eri
ali
simmer
sedi
nit
.
Al
l
owi
ttor
emai
nfor10-
15mi
nut
es.
4. Addaf
ewdr
opsoft
hest
ainont
hesmear
edmat
eri
al.Remov
ethel
argepi
eces
anddebr
is.Af
teraf
ewmi
nut
es,
freshdr
opsoft
hest
aincanbeadded.
5. Heatt
hesl
i
degent
lyov
ert
hef
lameandaddaf
ewdr
opsof20%acet
icaci
dor
uset
hest
aini
tsel
f.Cov
ert
hemat
eri
alwi
thacov
ersl
i
p.
6.Obser
vet
hesl
i
deundert
hemi
croscopeandi
dent
if
ythedi
ff
erentst
agesof

72
 mei
osi
s.

LABORATORYSTUDYNUMBER10

COLLECTI
ONANDPRESERVATI
ONOFPLANTANDI
NSECT
SPECIMENS

10.
1Int
roduct
ion
Col
l
ect
ionsofpl
ant
sand ani
mal
sser
veast
hesci
ent
if
icr
esear
cher
s,ar
ti
sts,and
educat
ors.Col
l
ect
ions may i
ncl
ude a v
ari
ety ofpr
epar
ati
on t
ypes emphasi
zi
ng
pr
eser
vat
ionofl
eaf
,st
em,
sof
tti
ssues,
teet
h,ski
norsomecombi
nat
ionoft
hese.

Obj
ect
ives:

Af
tercompl
eti
ngt
heact
ivi
ti
esst
udent
swi
l
lbe:
 abl
etodescr
ibeappr
opr
iat
etechni
quesf
orcol
l
ect
ingandpr
epar
ingher
bar
ium and
i
nsectspeci
mens
 begi
nthei
rownpl
antandani
mal
col
l
ect
ion
 acquai
ntedt
obasi
cequi
pmentusedf
ori
nsectcol
l
ect
ion
 acquai
ntedt
omet
hodsempl
oyedt
oki
l
landpr
eser
vei
nsect
s
 acquai
ntedt
ocor
rectpi
nni
ng,
mount
ing,
label
i
ngandst
ori
ngt
echni
ques

I
.Pl
antcol
l
ect
ing
Pl
antcol
l
ect
ingi
stheacqui
sit
ionofpl
antspeci
mensf
ort
hepur
posesofr
esear
ch,
cul
ti
vat
ion,orasahobby
.Pl
antspeci
mensmaybekeptal
i
ve,butar
emor
ecommonl
y
dr
iedandpr
essedt
opr
eser
vet
hequal
i
tyoft
hespeci
men.

Thepur
posesofcol
lect
ing:

73
Ther
ear
etwo mai
nreasonsf
orcol
l
ect
ing pl
ant
s.Thef
ir
sti
sto obt
ainr
ecor
dsand
speci
mensofpl
ant
s,ei
therf
oraper
sonalcol
l
ect
ionort
obest
oredi
naher
bar
ium.A
her
bar
ium speci
meni
sapr
essedand/
ordr
iedsampl
eofapl
antorf
ungust
hatcanbe
st
oredf
orf
utur
eref
erence.

Whatt
olookf
ori
naspeci
men:
Speci
mensf
orcol
l
ect
ionshoul
dbeascompl
eteaspossi
ble.I
deal
l
yfl
ower
sandf
rui
t
shoul
dbei
ncl
uded,
aswel
lasv
eget
ati
vepar
ts
Whatt
ocol
lect
/pl
antspeci
mens:
Eachspeci
menshoul
dconsi
stofast
em wi
that
tachedl
eav
esand,
ifatal
lpossi
ble,
f
lower
sand/
orf
rui
ts;
thesear
eusual
l
ycr
it
ical
fori
dent
if
icat
ion.Wi
thsmal
l
erpl
ant
s,
suchasgr
asses,
rushes,
sedges,
iri
sesandl
i
li
es,
col
l
ectwhol
epl
ant
s(oranumberof
ent
ir
epl
ant
s)i
ncl
udi
ngunder
groundpar
ts(
i.
e.bul
bs,
cor
ms,
rhi
zomes)st
il
lat
tachedt
o
aer
ial
par
tsofpl
ant
.Int
hecaseofv
eryl
arget
rees,
shr
ubs,
orv
ines,
piecesshoul
dbe
sel
ect
edt
oil
l
ust
rat
etot
hegr
eat
estext
entpossi
blet
heov
eral
lchar
act
eri
sti
csoft
he
pl
antandt
her
angeofv
ari
ati
oni
nfl
ower
s,l
eav
es,
andot
herst
ruct
ures.Make
speci
mensl
argeenought
opr
esentaf
airsampl
eoft
hepl
ant
,it
smannerofgr
owt
h,
br
anchi
ngandsoon.Mossesandl
i
chensshoul
dal
sobet
akenwhol
e.
Howt
ocol
lect
:
I
deal
l
y,col
l
ect
ionsofv
ascul
arpl
ant
sshoul
dbeputi
mmedi
atel
yint
oaf
iel
dpr
ess(
ify
ou
hav
etakenapl
antpr
essal
ong)
,becauset
hispr
oducest
hebestl
ooki
ngspeci
mens.
Col
l
ect
ingi
ntopl
ast
icbagsi
sanot
heropt
ionandt
hespeci
menswi
l
lbepr
essedl
ateron
af
tert
hef
iel
d.

Not
est
otake:
Ev
ery speci
men shoul
d be accompani
ed by compr
ehensi
ve not
es r
etai
ned i
na
col
l
ect
ingnot
ebook.Thenot
esshoul
dcont
aint
hef
oll
owi
ngi
nfor
mat
ion.
1.Col
l
ect
ionnumber
:Thi
sisaser
ial
numberspeci
fi
ctoacol
l
ect
orandaspeci
men.
Thenumbermayst
artat1andcont
inuet
hrought
hecol
l
ect
or'
sli
fet
ime.
2.Thenameoft
hepl
ant
:Thi
sisi
mpor
tantasi
thel
pst
hecol
l
ect
orr
emembert
he
i
ndi
vi
dualspeci
menev
eni
fthel
abel
sar
eacci
dent
lyl
ostormi
xed.Ev
eni
fthe
col
l
ect
orhasnoi
deawhatt
hespeci
meni
s,i
tissomet
imesusef
ult
ogi
vea

74
 compl
etel
yar
bit
rar
yname.
3.Local
i
ty:Thi
sshoul
dbeasdet
ail
edaspossi
ble,i
ncl
udi
ngt
henameoft
owns,
r
oadsl
akesandsooni
nthev
ici
nit
y,aswel
lasTownshi
p,Count
yorDi
str
ict
,the
l
ati
tudeandl
ongi
tude.
4.Descr
ipt
ion:Thi
sshoul
dincl
udeev
ery
thi
ngaboutt
hepl
ant
.Essent
iali
temsar
e
t
hehei
ght
,ty
peofbar
k,whet
hert
hest
em i
supr
ight
,spr
awl
i
ngordr
oopi
ng,
obv
ioussmel
l
s,whet
hert
hepl
anti
scl
umped,si
ngl
eorgr
owi
ngi
npat
ches,and
t
hepr
esenceofcr
eepi
ngorunder
groundst
ems.Fl
owerandf
rui
tcol
ourshoul
d
al
sobenot
edast
heseof
tenf
adeondr
iedspeci
mens.
5.Habi
tat
:Thi
sshoul
dincl
udet
hegener
alhabi
tataswel
lasmor
especi
fi
cdet
ail
s
ofmi
cro-
habi
tat
.Impor
tantpoi
ntsar
ety
peofsoi
lorot
hersubst
rat
e(sand,cl
ay,
gr
ani
te,deadwood,ot
herv
eget
ati
on)
,associ
atedspeci
es,moi
stur
eandaspect
(
ful
l
yexposedonasout
hfaci
ngbank;
inadamphol
l
owunderdensescr
ub,
etc)
.
6.Dat
eofcol
l
ect
ion
Toav
oidconf
usi
on,
wri
tet
hedat
einf
ull
(August7,
2018)
.Not
ethat‘
‘
8/07/
2018’
’
coul
dbei
nter
pret
edasAugust7,
2018orJul
y8,
2018.
7.Name(
s)ofcol
l
ect
or(
s)
Recor
dal
lsi
gni
fi
cantmember
soft
hecol
l
ect
ingpar
ty.I
nthef
utur
e,i
fone
col
l
ect
ori
sunav
ail
abl
etoanswerquest
ions,
theot
her
smaybecont
act
ed.

Pr
epar
ingpl
antspeci
mens
Af
tercol
l
ect
ionpl
ant
sar
epr
essedanddr
ied,
mount
edandst
ored.
1.Pr
essi
nganddr
yingpl
antspeci
mens
Speci
mensshoul
d bepr
essedwhenf
resh(
i.
e.i
nthef
iel
d).Thi
sresul
tsi
nbet
ter
her
bar
ium speci
mens,maki
ngt
hem easi
ert
oident
if
y.Whenpr
essi
ngaspeci
men,
car
eful
l
yspr
eadoutst
ruct
ures(
i.
e.l
eav
es,f
lower
s)sot
hatdi
agnost
icf
eat
uresar
e
cl
ear
lyev
ident
.Makesur
ethatbot
htheupperandt
hel
owerl
eafsur
facear
evi
sibl
eby
t
urni
ngov
ersomel
eav
es.Speci
mensar
epr
essedi
napl
antpr
ess,whi
chconsi
stsofa
wooden f
rame(
forr
igi
dit
y),cor
rugat
ed car
dboar
dvent
il
ator
s(t
o al
l
ow ai
rtof
low
t
hrought
hepr
ess)
,bl
ott
erpaper(
toabsor
bmoi
stur
e),andf
oldedpaper
,ty
pical
l
ya
newspaper(
tocont
aint
hepl
antmat
eri
al)
.Thepl
antpr
essi
sti
ght
enedusi
ngst
rapswi
th

75
buckl
esorbol
tswi
thwi
ngnut
s.Theobj
ect
iveofpr
essi
ngpl
ant
sist
oext
ractmoi
stur
e
i
ntheshor
testper
iodoft
ime,whi
l
epr
eser
vingt
hemor
phol
ogi
cali
ntegr
it
yoft
hepl
ant
,
andt
oyi
eldmat
eri
alt
hatcanber
eadi
l
ymount
edonher
bar
ium paper(
anaci
d-f
ree
car
dst
ock)f
orl
ong-
ter
m st
orage.

Fi
gur
e10.
1:Pl
antpr
esser
s

Howt
opr
essandpr
eser
vepl
ant
s
 Topr
esst
hespeci
men,
cleanupt
hepl
ant
.Br
ushof
floosesoi
landbl
otof
fmoi
stur
e.
 Ar
ranget
hepl
antonasheetofnewspaper
.Nextt
oit
,pl
acet
hei
dent
if
icat
iont
ag
wi
thi
tsname,
anumbery
ouhav
eassi
gnedt
oit
,thel
ocat
ionwher
eitwascol
l
ect
ed,
wheni
twascol
l
ect
ed,
andbywhom.Makesur
ethesamei
nfor
mat
ioni
siny
our
j
our
nal
.Pl
aceanot
herpi
eceofnewspaperont
opoft
hepl
ant
.
 Makel
ayer
s.Pl
acet
hepi
ecesofnewspaperwi
thy
ourspeci
meni
nsi
debet
weent
wo
pi
ecesofbl
ott
ingpaper
,thenbet
weent
wopi
ecesofcor
rugat
edcar
dboar
d,t
oal
l
ow
ai
rtoci
rcul
ate.
 Pl
acet
her
esul
ti
ngpackagei
nthepl
antpr
essandgent
lyscr
ewi
tdown.Asan
al
ter
nat
ive,
youcanhol
ditsecur
elyt
oget
herwi
thst
raps,
orpl
acesomeheav
y
obj
ect
s(books,
bri
cks)ont
op.
 Youcandr
ysev
eral
plant
sint
hepr
essatonet
ime.Eachshoul
dbear
rangedi
nthe
76
 samel
ayer
sasdescr
ibedabov
e.
 Checkt
hepl
ant
sev
eryt
woort
hreeday
s,andr
epl
acet
hedamppaper
swi
thdr
yones.
I
twi
l
ltakef
rom t
wot
ofourweeksbef
oret
hespeci
mensar
ecompl
etel
ydr
y.

Fi
gur
e10.
2:Pr
essi
ngofpl
ant
s

Pr
ocessi
ngdr
iedspeci
mensi
nvol
ves3maj
orst
eps:
 i
dent
if
icat
ion
 l
abel
prepar
ati
on
 mount
ing
 st
orage
I
dent
if
icat
ion:Speci
mensneed t
o be cor
rect
lyi
dent
if
ied.I
fyou cannoti
dent
if
yit
y
our
sel
forhav
eanydoubt
,sendadupl
i
cat
especi
ment
oat
axonomi
cexper
tfor
conf
ir
mat
ion.

Labelpr
epar
ati
on:Label
s(not
est
akendur
ingcol
l
ect
ion)f
ort
hedr
yspeci
menscanbe
pr
epar
edbef
oret
hepl
ant
sar
eident
if
ied,
recor
dingi
nfor
mat
ionknownatt
hatt
ime.The
genus/
speci
es and i
dent
if
iercan be t
yped i
nlat
er.Not
ethatanyi
nfor
mat
ion on
char
act
ersandf
iel
dobser
vat
ionst
hatcannotbeobser
vedf
rom t
hepr
essedspeci
men:
 Habi
tat
:incl
udeabr
iefdescr
ipt
ionofwher
ethepl
anti
sgr
owi
ng
(
e. Themedat
g. ri
andr
agr
assl
and;
grazedpaddock;
weedyr
oadsi
de,
etc.
)anda

77
 l
i
stofot
herpl
ant
sgr
owi
ngi
nassoci
ati
on,
ifknown.

 Habi
t:r
ecor
dthegr
owt
hfor
m(e.
g.t
ree;shr
ub;v
ine;her
b)andhei
ght(
e.g.dense
shr
ubt
o2met
ershi
gh;spr
awl
i
ngher
b).Fort
rees,r
ecor
dthebar
kty
peand
ext
ent(
e.g.r
oughbar
kupt
o2met
ersonmai
ntr
unk,
smoot
habov
e).Bar
kty
pei
s
especi
all
yimpor
t nEucal
anti ypt
us.Al
sor
ecor
dthecol
ouroff
reshst
ems,
leav
es,
f
lower
s(f
orpl
ant
s)orpi
l
eus,
sti
peandl
amel
l
ae(
forf
ungi
).
 Abundance:
numberofpl
ant
satsi
te.Fr
equencyi
nthear
ea(
rar
e,occasi
onal
,
f
requent
/commonorabundant
).
 Subst
rat
e:f
orbr
yophy
tes,
fungi
andl
i
chens,
incl
udeadescr
ipt
ionoft
he
subst
rat
eonwhi
cht
hespeci
meni
sgr
owi
ng.
 Foral
gae,
col
l
ect
ingl
abel
sshoul
dincl
udei
nfor
mat
iononwat
ert
ype(
sal
i
ne,
br
acki
shorf
resh)andqual
i
ty(
suchasmuddyorpol
l
uted)
,aswel
laswhet
hert
he
al
gai
sfr
eeorat
tachedt
oasubst
rat
e.

Mount
ingspeci
mens:Oncemat
eri
ali
spr
essedandt
hor
oughl
ydr
ied,i
tismount
ed
onher
bar
ium sheet
s.Thest
andar
dsi
zef
ort
hesesheet
sis420x297mm.Theyshoul
d
bemadef
rom st
if
f,aci
d-f
ree,paperorcar
dboar
dofgoodqual
i
tysot
hatt
heywi
l
lnot
t
urny
ell
oworcr
ackwi
thage.

St
orageofspeci
mens:I
nor
derf
orpl
antspeci
menst
obekepti
ngoodcondi
ti
onf
or
f
utur
eref
erence,
adequat
est
oragecondi
ti
onsar
eneeded.Thest
oragespaceshoul
dbe
kepti
nal
ow l
i
ght
,low humi
dit
yenv
ironment
.Thet
emper
atur
eoft
hest
oragespace
shoul
dbekeptcool
, ween50–650F.
bet

I
tisi
mpor
tantt
okeept
hest
oragespacef
reeofhar
mfulpest
.Iti
srecommendedt
o
pr
otectt
he speci
mens by sheat
hing t
he sheet
sin seal
ed pl
ast
ic bags.Var
ious
pest
ici
desmayal
sobeusedt
opr
otectt
hest
oragespacef
rom pesti
nfest
ati
on.I
fpest
i
nfest
ati
onhasal
readyoccur
red,t
hesampl
esshoul
dbef
rozenf
ort
hreet
ofourday
s.
Fr
eezi
ngnewaddi
ti
onsofpl
antsampl
esi
sasuggest
edpr
event
ati
vemeasur
eagai
nst
t
hei
ntr
oduct
ionofpestt
othest
oragespace.

78
Act
ivi
ty1:Pl
antcol
lect
ion
Eachst
udentwi
l
lcol
l
ect5pl
ant
swhi
chi
ncl
udegr
asses,shr
ubs,her
bs,andt
rees.The
pl
ant
swi
l
lbepr
essedanddr
iedandsubmi
tt
edt
othei
nst
ruct
orwi
thal
lnecessar
y
i
nfor
mat
ion.

I
I.I
nsectcol
lect
ion
I
ntr
oduct
ion:
Thebestwayf
orast
udentt
oknowandappr
eci
atei
nsect
sisbycol
l
ect
ing
al
lsagesofi
nsect
sfr
om di
ff
erenthabi
tat
satdi
ff
erentseasonsoft
hey
ear
.Shor
tvi
sit
s
t
oknownhabi
tat
smaynoty
iel
drepr
esent
ati
vespeci
esofi
nsect
s.Ther
efor
epr
olonged
sear
chi
nli
kel
yhabi
tat
swoul
ddef
ini
tel
yresul
tinsuccess.

Akeenst
udent
,inaddi
ti
ont
owhathe/
shel
ear
nsi
ncl
ass,woul
dcomet
oknow mor
e
abouti
nsect
sasar
esul
tofr
egul
arcol
l
ect
ionf
rom di
ff
erenthabi
tat
s.Ast
udentmay
l
ear
nal
otaboutt
hel
i
feanddai
l
yact
ivi
tyofi
nsect
sbysi
mpl
yobser
vingt
hem i
nthei
r
nat
uralhabi
tat
s,orhe/
shemaysy
stemat
ical
l
ycol
l
ectandpr
epar
ethem f
orper
manent
di
spl
ayandi
dent
if
icat
ion.Asar
esul
tofsuchact
ivi
tyast
udentwoul
ddev
elopmuch
mor
eint
eresti
ninsect
saswel
lasappr
eci
atet
her
olet
heypl
ayi
nnat
ure.

Somebut
ter
fl
iesandbeet
lesar
esi
mpl
ycol
l
ect
edf
ort
hei
rbeaut
y.Ot
heri
nsect
smaybe
col
l
ect
edt
odet
ermi
net
hespeci
est
hatmaybeofmedi
cal
,vet
eri
nar
yoreconomi
c
i
mpor
tance.St
il
lmanyot
heri
nsect
smaybecol
l
ect
edt
oknow t
hei
nsectf
aunaofa
gi
venar
ea.

A st
udentshoul
dther
efor
e know some oft
he met
hods and t
echni
ques t
hatar
e
empl
oyedf
orcol
l
ect
ingdi
ff
erentt
ypesofi
nsect
sfr
om di
ff
erenthabi
tat
s.I
naddi
ti
on,
t
he st
udenthast
o be abl
eto car
eful
l
ypr
ocess,mount
,pr
eser
ve,i
dent
if
y(wher
e
possi
ble)andl
abel
his/
hercol
l
ect
ion.
I
twi
l
lbedi
ff
icul
tint
hisi
ntr
oduct
orycl
assorl
abor
ator
ysessi
ononent
omol
ogi
cal
t
echni
que t
o l
i
st al
lty
pes of met
hod f
or col
l
ect
ing and mount
ings i
nsect
s.
Ent
omol
ogi
calt
echni
quesact
ual
l
yinv
olv
esev
eralot
hermat
ter
s.Thest
udenti
sadv
ised

79
t
oconsul
tvar
iousbooksandmanual
stol
ear
nmor
eaboutt
hev
ari
ousmet
hodsusedi
n
t
hecol
l
ect
ionandpr
epar
ati
onofi
nsect
s.St
andar
dident
if
icat
ionkey
sorev
ensi
mpl
e
key
sthatar
efound i
n books coul
d be used t
oident
if
yinsect
stot
he f
ami
l
y.
I
dent
if
icat
ionofspeci
menst
othegenusort
hespeci
esi
susual
l
yver
ydi
ff
icul
tandmay
r
equi
ret
hehel
pofaspeci
ali
st

Wher
etol
ookf
ori
nsect
s
I
nsect
sar
efoundal
mostev
ery
wher
e.Mi
l
li
onsofi
nsect
scoul
dbecol
l
ect
edf
rom a
smal
lpl
otofl
and.Theyar
efound underst
ones,l
ogs,f
all
en l
eav
es and r
ott
ing
v
eget
ati
on.Al
argenumberofi
nsect
sar
efoundont
hef
lower
s,l
eav
es,br
anchesand
st
emsoft
reesorondi
ff
erentpar
tsofsmal
lpl
ant
sorcul
ti
vat
edcr
ops.Manyar
efound
i
nsoi
l
,and among gr
ass.Numer
ousi
nsect
sful
l
yorpar
ti
all
yli
vei
ntempor
aryor
per
manentbodi
esofwat
er.Adul
tandi
mmat
urest
agesofi
nsect
stend t
oli
vei
n
di
ff
erenthabi
tat
s.Fori
nst
ance,t
hel
araeandpupaeofCul
v exqui
nquef
asci
atusl
i
vei
n
dr
ainagesy
stems,sept
ict
anks,et
c.,whi
l
ethewi
ngedadul
tmosqui
tof
li
esi
ntonear
by
houses,
cav
esandnest
stof
eedonbl
oodofman,
var
iousr
odent
sorbi
rds.

Somei
nsect
sar
eact
ivedur
ingt
heday(
diur
nal
)whi
l
eot
her
stendt
obeact
iveatni
ght
(
noct
urnal
).Ther
efor
e,t
hehabi
tat
saswel
last
het
imeoft
hedayar
eimpor
tantt
o
col
l
ectdi
ff
erentst
agesandspeci
esofi
nsect
s.

Basi
cequi
pmentf
orcol
lect
ingi
nsect
s
1.I
nsectnet
s
Aer
ialnet
s:Thesear
eusedt
ocol
l
ecti
nsect
sinf
li
ght
.Anetbagmadeoft
ransl
ucent
net
ti
ngsoonecanseewhat
’si
nsi
de;
itcanbeusedasabeat
ingneti
fneeded.
Beat
ing(
orsweepi
ng)net
-Aheav
ycl
othbag,per
hapswi
thsmal
lnet
tedar
eaatt
he
bot
tom;
iti
susedt
osweep“
li
keabr
oom”t
hroughv
eget
ati
onmanyt
imes.I
tisusedf
or
t
aki
ngi
nsect
sthatl
i
veamongv
eget
ati
on.
Aquat
icnetorwat
ernet
:Iti
susedt
ocol
l
ecti
nsect
sfr
om aquat
ichabi
tat
.

2.Aspi
rat
ors

80
At
ubepl
uggedwi
thar
ubbercor
kinwhi
char
einser
tedt
wot
ubes:onebentandusedt
o
poi
ntatt
inyi
nsect
s;t
heot
herconnect
edt
oar
ubbert
ubef
ori
nhal
i
ngqui
ckl
ytosuck
t
hei
nsecti
ntot
het
ube.Thel
att
eronehasat
inyscr
eenat
tachedt
othei
nsi
deendt
o
pr
eventi
nsect
sfr
om get
ti
ngi
ntoone’
smout
h.Aspi
rat
orsar
eusedf
orcat
chi
ngsmal
l
i
nsect
ssuchasmosqui
toes,
sandf
li
es,
blackf
li
esmi
dges,
etc.

Fi
gur
e10.
3:Aspi
rat
or
3.Ber
lesef
unnel
Acommer
cialf
unnelofanysi
zei
sneeded,equi
ppedwi
thascr
eeni
nser
tedj
ustabov
e
t
henar
rowspoutt
opr
eventmat
eri
alf
rom f
all
i
ngout
.Leafl
i
tter
,bi
rds’nest
s,andot
her
or
gani
cmat
terar
eputi
ntot
hef
unnel
,whi
chi
smount
edonar
ackorr
ingst
and.Al
i
ght
bul
bispl
acedov
ert
het
opt
odr
youtt
heor
gani
cmat
eri
al,
dri
vi
ngar
thr
opodsdownwar
d
ast
heyseekmoi
stur
e.Thi
sisasi
mpl
emet
hodofcol
l
ect
ingi
nsect
s(e.
g.spr
ingt
ail
s)
outofsoi
l
,leafl
i
tter
,et
c.Ast
hemat
eri
aldr
ieswi
thel
ect
ri
cli
ghtbul
bthei
nsectmov
es
downt
hef
unnel
andget
scol
l
ect
edi
naj
arcont
aini
ng70%al
cohol
.TheBer
lesef
unnel
is
l
efti
npl
aceunt
ilt
heor
gani
cmat
teri
scompl
etel
ydr
iedout
.

4.Li
ghtt
raps
Var
ioust
ypesofl
i
ghtt
raps(
e.g.CDCl
i
ghtt
rap,
NewJer
seyl
i
ghtt
rap,
Monk’
sWoodt
rap,
et
c.)ar
eusedt
ocol
l
ectsmal
linsect
ssuchascul
i
coi
des,sandf
li
es,mosqui
toesand

81
bl
ackf
li
es.Li
ghtt
rapsar
enor
mal
l
yusedatni
ghtandar
eef
fect
ivei
ncat
chi
ngi
nsect
s
t
hatar
eact
iveatni
ght
.

5.Fl
oat
ati
ont
echni
que
Somei
nsect
sandr
elat
edar
thr
opodscanbecol
l
ect
edf
rom soi
l
,debr
isorl
eafl
i
tter
usi
ngt
hist
echni
que.Sol
uti
onofsugarormagnesi
um sul
phat
ecoul
dbepl
acedi
na
bucket
.St
irorshakeasampl
eofsoi
lint
hissol
uti
onori
nwat
er.Var
iousmi
tes,l
arv
ae,
pupaeandadul
tsofi
nsect
scoul
dfl
oatt
othesur
facef
rom wher
etheycoul
dbeeasi
l
y
r
emov
edusi
ngcamel
-hai
rbr
ushorf
orceps.
6.St
ickypl
ates
Thesear
efor
micapl
atesorsi
mpl
enon-
absor
bentpaperpl
atesofknownsi
zet
hatcoul
d
besmear
edwi
thst
ickymat
eri
alsuchascast
oroi
l
.Thepl
ateswi
l
lbel
eftov
erni
ghti
n
l
i
kel
yhabi
tatt
ocat
chorent
rapsmal
li
nsect
ssuchassandf
li
esandant
s.
7.Humanbai
t
Human bei
ngs coul
d ser
ve as bai
tto col
l
ect ant
hropophi
l
ic speci
es and
haemat
ophagousi
nsect
s(e.
g.mosqui
toes,
sandf
li
es,
blackf
li
es,
etc.
).

I
nsectki
ll
ingj
ars/
bot
tl
es
Agl
assorpl
ast
icj
arofdesi
redsi
ze(
tocol
l
ectsmal
landl
argei
nsect
s)canbemade
i
ntoaki
l
li
ngj
arbyput
ti
ngabouta1-
cm l
ayerofpl
ast
erofPar
isi
nthebot
tom,oruse
j
ustapadofabsor
bentmat
eri
alsuchascot
tonorsof
tti
ssue.Af
lui
dki
l
li
ngagentsuch
aset
hylacet
atei
saddedt
obeabsor
bedbyt
hepl
ast
erorot
hermat
eri
al.Besur
enott
o
hav
eanyf
lui
dont
hewal
l
soft
hej
ar,orspeci
menswi
l
lbespoi
l
ed.I
fyouusecot
tonor
ot
herabsor
bentmat
eri
al,cutacar
dboar
ddi
skt
osepar
atet
hei
nsect
sfr
om t
hepadof
ki
l
li
ngagent
.

1.Cy
ani
debot
tl
es(
Sodi
um orPot
assi
um cy
ani
de)
Toxi
ccy
ani
dev
apor
sar
eusedcommonl
ybyent
omol
ogi
stst
oki
l
ladul
tinsect
s.Cy
ani
de
i
sani
nsect
ici
dewhi
chi
sadeadl
ypoi
son.Sodi
um orpot
assi
um cy
ani
decov
eredwi
th
sawdustandpl
ast
erofPar
isi
nki
l
li
ngbot
tl
eswoul
dser
vei
ntheki
l
li
ngofdi
ff
erent
speci
esofi
nsect
s.Thi
sar
rangementshoul
dbepr
epar
edi
nat
ight
lycor
kedbot
tl
e.

82
Cal
cium cy
ani
debot
tl
eispr
epar
edi
nadi
ff
erentway
.Her
einst
eadofpl
ast
erofPar
is,
cot
tonandcar
dboar
dar
eusedt
ocov
ert
hecy
ani
dei
ntheki
l
li
ngbot
tl
e.

2.Et
hylacet
atebot
tl
e
Abot
tl
epar
tl
yfi
l
ledwi
thpl
ast
erofPar
iscoul
dbeusedf
ort
hispur
pose.Dr
ythepl
ast
er
andt
hensat
urat
eitwi
thet
hylacet
ate.Thi
ski
l
li
ngbot
tl
emustbekeptt
ight
lycor
ked.
Theet
hyl
acet
atewi
l
lremai
nef
fect
ivef
ormont
hs.

Fi
gur
e10.
4:Aki
l
li
ngj
ar

3.Ot
herki
ll
ingagent
s
Et
her
,chl
orof
orm,car
bont
etr
achl
ori
deandbenzenear
eal
sousedbycol
l
ect
orst
oki
l
l
i
nsect
s.

Mount
ingandpr
eser
vingi
nsect
s
Oncet
hespeci
mensar
edead,y
oucanmountt
hem.Remembert
otaket
hem outas
soonast
heyar
edead.Leav
ingt
hem i
nthej
art
ool
ongwi
l
ldamaget
hei
rcol
or.I
nsect
s
ar
eusual
l
ymount
edont
her
ightsi
de(
wheny
oul
ookati
tfr
om behi
nd)
.
83
1.Pi
nni
ng

AGui
det
omount
ingi
nsect
sonpi
ns

Thegr
oupsnor
mal
l
ymount
edoni
nsectpi
nsar
e:Odonat
a,Or
thopt
era,Der
mapt
era,
Hemi
pter
a(Het
eropt
era)
,Homopt
era(
Auchenor
rhy
ncha)
,Neur
opt
era,Col
eopt
era,
Mecopt
era,Lepi
dopt
era,Di
pter
a,andHy
menopt
era.Mostsmal
linsect
sthatcanbe
pi
nnedar
eatl
east5mm i
nlengt
h,wi
that
hor
axbi
genought
ohol
dthepi
n.Most
medi
um andl
argei
nsect
sar
epi
nnedent
omol
ogi
calpi
nsofsi
ze1t
o3.Si
zes4t
o7ar
e
somet
imesav
ail
abl
eforl
argespeci
mens.

Fi
gur
e10.
5:Av
ari
etysi
zeofent
omol
ogi
cal
pins
 Beet
les-pi
nont
heupperquadr
antoft
her
ightwi
ng(
ely
tra)
 Wasps,
bees,
andf
li
es-pi
ntot
her
ightoft
het
hor
ax,
neart
hebaseoft
hewi
ngs.
 Mot
hsandbut
ter
fl
ies-pi
nint
hemi
ddl
eoft
het
hor
ax;
thi
sist
obal
anceoutt
he
i
nsect
sforspr
eadi
ngt
hei
rwi
ngs.
 Gr
asshopper
s-pi
ntot
her
ightoft
het
hor
ax.
 Dr
agonf
li
esandDamsel
fl
ies-ar
enotusual
l
ypi
nned.Theyar
ekepti
nenv
elopes.
Poi
nti
ng:I
nsect
sthatar
etoosmal
ltobepi
nnedar
emount
edonpoi
ntsmadewi
th
paper
.Make
t
ri
angul
arpoi
ntsoutofanon-
aci
dpaper(
suchasar
ti
stcanv
as)
.Thepoi
ntsshoul
dbe

84
 onl
y7-
10mm l
ongandt
hewi
dthoft
het
hickestend2-
3mm wi
de.

Di
agr
am ofpi
nni
ngl
ocat
ions

85
 A.Fl
iesandbees
B.Butterf
li
esandmot hs
C.Grasshoppersandcri
ckets
D.Truebugs
E.Dragonfli
esanddamselfl
i
es
F.Beetlesandweevil
s

Fi
gur
e10.
5:Cor
rectpi
nni
ngl
ocat
ionofsomei
nsectgr
oups

Lar
geI
nsect
s

86
 Pi
nthei
nsect
,dor
salsi
deup,sot
hatpr
eci
sel
y10mm oft
heshaf
tisf
reeabov
ethe
speci
men.Gener
all
y,t
hepi
nshoul
dpasst
hrought
hei
nsect
'smesot
hor
ax,sl
i
ght
lyt
o
t
her
ightofcent
er(
seeDi
agr
am ofPi
nni
ngLocat
ionsf
ordi
ff
erentbodyt
ypes)
.The
i
nsect
'sv
ent
ralsur
faceshoul
dbeper
pendi
cul
art
othepi
n.Consi
derv
isi
bil
i
ty,
breakage,
andspacewhenposi
ti
oni
ngpar
ts.Lepi
dopt
eraandOdonat
aar
ebestposi
ti
onedona
spr
eadi
ngboar
d.Wi
thLepi
dopt
eraspr
eadsot
hatt
het
rai
l
ingedgeoft
hef
orewi
ngj
ust
ov
erl
apst
hel
eadi
ngedgeoft
hehi
ndwi
ng,t
hel
i
neofj
unct
urebei
ngper
pendi
cul
art
o
t
hel
ongi
tudi
nal
axi
soft
hebody
.

Fi
gur
e10.
6:Cor
rectpi
nni
ngofl
argei
nsect
sli
keOr
thopt
era

Smal
lInsect
s

I
nsect
stoosmal
ltor
ecei
vet
heshaf
tofa#3pi
nmaybegl
uedt
opi
nnedpaperpoi
nts.Usi
ngcl
ear
nai
lpol
i
shasgl
ue,f
ir
mlyat
tacht
hepoi
ntt
othei
nsect
'sr
ightsi
dej
ustabov
ethemi
ddl
eri
ghtl
eg
(
mesot
hor
ax)
,dor
salsi
deup.Al
i
gnl
abel
s5mm apar
tasshownt
opr
otectt
hespeci
menand
conser
vespace.

87
Fi
gur
e10.
7:Cor
rectpi
nni
ngofsmal
linsect
sli
kesmal
lant
s

Dat
alabel
s

Usei
ndexcar
dst
ockorequi
val
ent
.Label
sshoul
dbel
ett
eredi
nbl
ack,
wat
erpr
oofi
nkor
pr
int
edwi
thal
aserpr
int
er.Acr
owqui
l
lorRapi
dogr
aphpen(
01poi
nt)i
sadequat
efor
mosthandl
ett
eri
ng.
Si
ze:
nol
argert
han8x12mm,
1-4l
i
nesofwr
it
ingperi
ndi
vi
dual
label
.
For
mat
:Wr
it
edat
a(wher
e,when,
andbywhom col
l
ect
ed)

1.Topl
evel
:Dat
e/l
ocal
it
ylabel(
gener
alt
ospeci
fi
c)
Li
nes1&2:Count
ry,
Stat
e,Count
y,Ci
ty

Li
ne3:Dat
ecol
l
ect
ed(
day
,mont
hinRoman,
yeari
nful
las26-
VII
-2011)
Li
ne4:Col
l
ect
or'
sname(
ini
ti
alsandl
astname)

2.Secondl
evel
:Ident
if
icat
ionl
abel
 Gi
vef
ami
l
ynamef
orpi
nnedi
nsect
s;or
derandf
ami
l
yfori
nsect
sinal
cohol

88
 3. Thi
rdl
evel
(opt
ional
):Ecol
ogi
call
abel

Medi
um andl
arge-
sizedi
nsect
ssuchasbut
ter
fl
ies,beet
les,andgr
asshopper
scanbe
pi
nned.Thepi
nsshoul
dbet
her
ightsi
zeandshoul
dnotbepi
nnedont
hepar
toft
he
bodyt
hatwoul
dnotmakei
dent
if
icat
iondi
ff
icul
t.Thel
abel
sar
eat
tachedt
othepi
nsor
sl
i
deswi
thmount
edspeci
mens.Wi
thpr
eser
vedspeci
menst
hel
abel
sar
ewr
it
teni
n
penci
landkepti
nbot
tl
eswi
tht
hespeci
mensandt
hepr
eser
vat
ive(
e.g.70% or80%
al
cohol
).

2.Ot
hermet
hodsofmount
ing
Ver
ysmal
li
nsect
sar
emount
edonacar
dpoi
ntoronmi
croscopi
csl
i
des.
3.Spr
eadi
ngi
nsect
s
I
nsect
ssuchasbut
ter
fl
iesar
epr
epar
edonspr
eadi
ngboar
dbef
oret
heyar
ereadyf
or
di
spl
ay.

Pr
eser
vingi
nsectspeci
mensi
nal
cohol
I
nsect
sthatar
etoosmal
l(e.
g.smal
lbeet
les,
ant
s,manybugs,
etc.
),ort
hebodi
esof
whi
char
etoobr
it
tl
eorsof
t,shoul
dnotbepi
nned.Theyar
eki
l
ledandst
oredi
ngl
ass
v
ial
sin70%or80%et
hanol
.

Rel
axi
ng
Fr
eshspeci
menofi
nsect
scaneasi
l
ybemount
ed.Dr
iedspeci
menar
edi
ff
icul
ttomount
caneasi
l
ybr
eak.suchspeci
menar
erel
axedbyput
ti
ngt
hem i
nrel
axi
ngchamber
cont
aini
ngwetsandorcl
othf
oradayort
wo.Car
bol
i
caci
disaddedt
opr
eventmoul
d.

I
nsectboxes
Pr
epar
edspeci
mensar
ekepti
nboxes(
lar
georsmal
l
)topr
otectt
hem f
rom damageor
i
nsectpest
s.Label
edspeci
menskepti
ninsectboxar
eei
therst
oredi
nasaf
epl
acef
or
f
utur
euseorar
eputondi
spl
ayf
orusebyst
udent
sort
hegener
alpubl
i
c.

Act
ivi
ty2:I
nsectcol
lect
ion

89
Col
lect
ioni
nst
ruct
ions

Eachst
udentshoul
dsubmi
tani
nsectcol
l
ect
ionf
rom t
en(
10)or
der
s.Thecol
l
ect
ion
mustbehousedi
nSchmi
ttboxesorshi
rtboxes,andspeci
mensmustbemount
edon
i
nsectpi
ns(
notor
dinar
yhousehol
dst
rai
ghtpi
ns)orpr
eser
vedi
nvi
alsofal
coholi
fsof
t-
bodi
ed.Eachpi
nnedi
nsectandeachv
ialofal
coholmustcont
ainadat
e/l
ocal
i
tyl
abel
whi
chi
ncl
udest
hef
oll
owi
ngl
abel
i
ngi
nfor
mat
ion:
1.nameofspeci
mens
2.pl
aceofcol
l
ect
ion(
local
i
ty)
3.dat
eofcol
l
ect
ion
4.nameofcol
l
ect
orofspeci
mens

90
 REFERENCES
r
d
Tay
or,
D.J.
,Gr
een,
N.P.
O.andSt
out
,G.W.U.(
1998)
.Bi
ologi
calSci
ence3 edi
ti
on.Pp
984(
Soper
,R.
,ed)
.Cambr
idgeUni
ver
sit
yPr
ess.

Ht
tp:
//www.
sspca.
org/
gener
albi
ology
.ht
ml.Sear
chedonSept
ember5,
2017.

91
 Appendi
x

Labor
ator
yRepor
tWr
it
ingFor
mat

Thepur
poseofwr
it
ingal
abr
epor
tist
odet
ermi
nehow wel
lyouper
for
medy
our
exper
iment
, how much y
ou under
stood about what happened dur
ing t
he
exper
iment
ati
onpr
ocess,
andhowwel
lyoucanconv
eyt
hati
nfor
mat
ioni
nanor
gani
zed
f
ashi
on.

LabRepor
tFor
mat
Agoodl
abr
epor
tfor
mati
ncl
udessi
xmai
nsect
ions:
1.Ti
tl
e
2.I
ntr
oduct
ion
3.Mat
eri
als
4. Met
hods
5.Resul
ts
6.Di
scussi
on
7.Concl
usi
on
8.Ref
erences

Keepi
nmi
ndt
hati
ndi
vi
duali
nst
ruct
orsmayhav
easpeci
fi
cfor
matt
hatt
heyr
equi
rey
ou
t
ofol
l
ow.Pl
easebesur
etoconsul
tyourt
eacheraboutt
hespeci
fi
csofwhatt
oincl
ude
i
nyourl
abr
epor
t.

Ti
tl
e
Thet
it
lest
atest
hef
ocusofy
ourexper
iment
.Thet
it
leshoul
dbet
othepoi
nt,
descr
ipt
ive,
accur
ate,
andconci
se(
tenwor
dsorl
ess)
.Int
hiscour
sey
ouuset
hegi
vent
it
lef
oreach
act
ivi
tyi
nthemanual
.
 Ti
tl
epage

92
Notal
ll
abr
epor
tshav
eti
tl
epages,
buti
fyouri
nst
ruct
orwant
sone,
itwoul
dbeasi
ngl
e
paget
hatst
ates:
 Thet
it
leoft
heexper
iment
 Yournameandt
henamesofanyl
abpar
tner
s
 Youri
nst
ruct
or'
sname
 Thedat
ethel
abwasper
for
medort
hedat
ether
epor
twassubmi
tt
ed

I
ntr
oduct
ion
I
tisusual
l
ysev
eralpar
agr
aphsl
ong(
butshoul
dnotbet
ool
engt
hy)
.Thei
ntr
oduct
ionof
al
abr
epor
tst
atest
hepur
poseofy
ourexper
iment
,whyt
hest
udywasunder
taken
(
obj
ect
ivesoft
hest
udy
);gi
veabr
iefsummar
yofr
elev
antbackgr
oundi
nfor
mat
ion;
and
endwi
thast
atementoft
hespeci
fi
cpr
obl
em y
ouexami
ned.

Mat
eri
als
Li
stev
ery
thi
ngneededt
ocompl
etey
ourexper
iment(
Mat
eri
alsandchemi
cal
s).

Met
hods
Descr
ibet
hest
epsy
oucompl
eteddur
ingy
ouri
nvest
igat
ion.Thi
sisy
ourpr
ocedur
e.Be
suf
fi
cient
lydet
ail
edt
hatany
onecoul
dreadt
hissect
ionanddupl
i
cat
eyourexper
iment
.
Wr
it
eitasi
fyouwer
egi
vi
ngdi
rect
ionf
orsomeoneel
set
odot
hel
ab.

Resul
ts
Ther
esul
tssect
ionshoul
dincl
udeal
ltabul
ateddat
afr
om obser
vat
ionsdur
ingy
our
exper
iment
.Thi
sincl
udeschar
ts,t
abl
es,gr
aphs,andanyot
heri
l
lust
rat
ionsofdat
ayou
hav
ecol
l
ect
ed.Youshoul
dal
soi
ncl
udeawr
it
tensummar
yoft
hei
nfor
mat
ioni
nyour
char
ts,t
abl
es,and/
orot
heri
l
lust
rat
ions.Anypat
ter
nsort
rends obser
ved i
nyour
exper
imentori
ndi
cat
edi
nyouri
l
lust
rat
ionsshoul
dbenot
edaswel
l
.Somet
imest
he
Resul
tssect
ioni
scombi
nedwi
tht
heDi
scussi
on(
Resul
ts&Di
scussi
on)
.

Di
scussi
on
Thi
siswher
eyoui
nter
prety
ourdat
a/r
esul
ts.I
tisal
sowher
eyouwoul
ddi
scussany

93
mi
stakesy
oumi
ghthav
emadewhi
l
econduct
ingt
hei
nvest
igat
ion.I
ndi
scussi
ony
ou
conv
incey
ourr
eadert
haty
ouar
eint
erpr
eti
ngt
hedat
aful
l
yandi
ntel
l
igent
ly.Whatdi
d
y
oul
ear
n?Wer
ether
eanyer
ror
s?LongerDi
scussi
onov
er1-
2pagesi
snotal
way
sbet
ter
!

Concl
usi
on
Mostoft
het
imet
heconcl
usi
oni
sasi
ngl
epar
agr
apht
hatsumsupwhathappenedi
n
t
heexper
iment
,whet
hery
ourhy
pot
hesi
swasaccept
ed orr
eject
ed,and whatt
his
means.Somet
imesConcl
usi
oncanbemer
gedwi
thDi
scussi
onas‘
Discussi
onand
Concl
usi
on’
.

Ci
tat
ion/
Ref
erences
Al
lref
erencesusedshoul
dbei
ncl
udedatt
heendofy
ourl
abr
epor
t.Ref
erencesi
ncl
ude
anybooks,
art
icl
es,
labmanual
s,et
c.t
haty
ouusedwhenwr
it
ingy
ourr
epor
t.

94