TheMicroscope

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The microscope,developedmorethanthreecenturiesago,is the basictool of the biologist.A

microscopeenablesbiologistsio investigateliving things and objectsthat aretoo smallto be seenwith the
unaidedeye.The microscopeis ableto magnifr thesetiny specimensby meansof lenseslocatedin the
eyepieceandobjectives.T[e [gtrt microscopeis also capableof revealingfine detail. This ability to revealfine
aitiit is known asresolvingpower. The type of microscopethat you will be usingthroughoutyour studyof
biology
- is the compoundlight microscope.
Specimensihat areviewedunderthe microscopearemountedon one of two typesof glassslides.
Preparedslidesare slidesthat are for permanentuse.\ilet-mount slidesare for temporaryuse.Most of the
slidesthat you will usein biologywill be wet mounts.
for youto estimatethe lengthandwidth of someolfg,ur
In ybur studyofbiology, it will be necessary
specimens.To measureobjecti underthe microscope,a unit calledthe micrometer(pm), rygetimes called
tfremicton (p), is used.Onemicrometerequals0.001millimeter;onemillimeterequals1000micrometers.
Give the purposeof the microscope:
Defineresolvingpower:
Namethetype of microscopeusedin high schoolbiologyplasses:
Contrastthe useof preparedandwet-mountslides:
for microscopework:
Namethe preferredunit of measurement
Tell how manymicrometersare in a millimeter:

alsocalledthe

Give the syrnbolfor the micrometer:

Stage opening
permitslight from light sourceto
pass uP through body tub

Oiaphngm
regulatesamount ol l.lght
passingthrough sPeclmen
Lamp or mirtor
directs lighl through diaphragm
and stage openrng

Carrying the Microscope: Takethe microscopefrom the storagearca. Carrythe microscopewith onehand
underthe baseandthe otherhandgraspingthe arm.
Placethe microscopeon the laboratorytable. The microscopeshouldbe about 10 cm from the edgeof the
table. Uncoil andplW the cord into anoutlet at your lab station. The on(l)-otr(0) switchis locatedon the
left sideofthe baseofthe scope.
Parts of the Microscope: Look at the drawingson the previouspagethat is mostlike your microscope.
Identifr the partsand functionsof your microscope.
Cleaningthe Microscope: Carefullycleanthe eyepieceandobjectiveslenseswith lenspaper. Locatethe
nosepieceand gently turn it sothat the low-powerscanningobjectiveis in line with the body tube. The
nosepiecewill click into placewhen the objectiveis in the properposition.
Field of View: Keepingboth eyesopeo look throughthe eyepiece.You will seea circleof light. This is
calledthe field of view. To makethe circle of light asbright aspossible,you may haveto adjustthe
diaphragm.
Your Dominant Eye: If you areobservingthrougha singlelens(monocular)scope,learnto seewith your
dominanteyewhile training the other eyeto relax andnot concentrateon anything. This techniqueis useful
and is easily learnedwith practice. It will comein handywhenyou draw your specimensasyou canuseyour
weakereyeto help you seeyour drawingwhile observingwith the dominanteye.
Peoplenormally havea stronger
(dominant)eye(the onethat is
mostused). It is oftenthe one
usedfor microscopicobservation,
unlessyou areusing a binocular
stereomicroscope.Then,you
would useboth eyesto observe
your objector specimen.

Right or Left? Whichis your dominanteye?

In orderto determinewhich eyeis dominant(stronger),thereis a
simpletest. Useyour handsto form a circleat arm'slength. Look
acrossthe roonl throughthe circle,both eyesopen(relax),at some
object. Slowly bringthe circletowardyour face. Your "handcircle" will probablygo to oneeyeor the other-this is your
dominanteye. Usethis eyefor viewing.

Describeyour hand-positionwhen
carryingthe microscope:
Give the rulmefor the circle of light you seewlren looking throughthe eyepiece:
Namethe microscopepart usedto adjustthe amountof light in the field of view:
Contrasta monocularand binocularscope:
Which eye is your dominanteye?
Therearethreeobjectivelenseson your microscope.The shortestlensis calledthe scanninglens-it hasa
red ring aroundit. The low-powerobjectiveis the medium-lengthlensand hasa yellow ring aroundit. The
high-powerobjectiveis the longestlensand hasa blue ring aroundit.
The lensyou areusing is the onethat is "clicked" into placeabovethe stageopening. You could also sayit
is the onebelowthe bodytube.
A4iust your micrqscopesothe scanninglgnsis in placefor viewine a specimen.

Preparing and Observing a \ilet Mount

Obtain a glassslide and cover slip. Wipe both sidesof the slidewith a cloth to removedust,etc. The cleaned
slide andcover slip shouldbe handledby their edges.
Cut out a smallpieceof newspaperthat containsthe letter "e." This shouldbe a standardnewsprintletterNOT a headline-sizeletter-Avoid dark backgroundson either sideof the newsprint. Placethe letter "e" in the
centerofthe slide as it would appearin the newspaper-face-up.rieht sideup.
Usinga pipette,placea dropof tap wateron top ofthe "e." IJsea probeto hold the "e" in placewhile you add
the drop of water,if necessary.
overthe dropof water.Gentlylower the coverslip ontothe slide. If air
Hold a coverslip at abouta45o ang,le
gently
tap the coverslip with the backendof the probe.
bubblesappear,

Dissectingneedle

Placethe wet mountof the letter "e" on the stageof the microscoperryiththe letter facing yor+asyou would
readit in print. Adjust the slide so that the letter is abovethe openingof the stage.
Look at the slide at eyelevel. Observethe spacebetweenthe
slideandthe scanningobjective.The scanningobjectiveshould
be in line with the body tube (over the stageopening).Slowly
knob, raisingthe stageto its highest
turn the coarse-adjustment
position. You arenow readyto view your specimen.
Look throughthe eyepieceandSLOWLY turn the coarseadjustmentknobuntil the letter"e" comesinto focus. Inthe
a field of view), sketchwhatyou
circlebelow(whichrepresents
see.
Calculatethe magnification(eyepiecex objective).
Showyour math in the spacebelow.

Leaveyour slide on the stageandhaveyour instructorveriff

your view. Your instructorwill initial the blank below.
Magnification
Completed

CAUTIONz Never raise the stage

while looking through the eltepiece;
you may hit and danrage the slide or
ohjective lens. Look to the side, raise
the stage to its nruxintumheight, then
focus u,hile the stage is movirtgAWAY
fi'om the objective.

Make a secondslideof a lettero'e"but this time from print providedby your instructor.Onceagain,checkthe
orientationof the letter(it shouldbe in the samepositionasit appearsin print).
Rotatethenosepiece
to the actuallow-powerlens. This is the mid-lengthlens(with a yellow circle)andis
markedwith a magnificationof 10X.
Followingpropertechnique,locateandfocuson the letter. Sketchwhatyou observebelow.
Calculatethe magnificationbelow(showyour math):

Completed

Magnification

ParadoxicalMovement: Themicroscope
exhibitsan opticalphenomenon
calledparadoxical
movement.
Movethe slideto theright......Whichway doesit "appear"to movein the field of view?
to theleft....Whichway doesit o'appear"
to move?
Move the slideawayfrom you,
you. Which way doesit "appear"to moveeachtime?

Move it
thentoward

Whatdoesit mean

whenan opticalinstrumentexhibitsparadoxicalmovement?
Practicecenteringan objectin the field of view. For example,an objectis on the edgeof the field of view and
you wantto centerit beforeyou askyour partnerto takea look. Determinewhich way you would movethe
slideto centerthe specimen.Below aresomesampleproblems.Usean arrow(or two arrows)to showwhich
way you would movethe slideto centerthe object:

To observea specimenat high-power magnifrcation,turn the nosepieceuntil the high-power objective clicks
into place. (The high-powerobjectiveis the longestobject,has a blue ring, and is marked40X.)
You microscopeis parfocal. This allows you to focus the scopeat low power, switch to high power, and,with
only minor adjustmentof the fine-adjustmentknob, seethe object at high power. For that reason,here is
anothergeneralrule of microscopy:
To view an object at high power, always begin by focusing on it at low power. Then,
switch to high power and use the fine-adjustment knob to bring the object into view.
Observethe letter at high power. Draw what you observe
Calculatethe magnificationbelow (show your math):

Completed

Magnification

Measuringan ObjectUndertheMicroscope
Turn the nosepieceso the scanninglens (4x, 40x total) is in place. Placea millimeter scaleof a transparent
plastic ruler over the centerof the stageopeningin the microscope.
Use the scanningobjectiveto locatethe millimeter lines of the ruler. Placetheselines in the middle of the field
of view and use the coarse-adjustment
knob to bring them in to focus. The distancebetweentwo lines on the
ruler representsI mm.
While looking through the eyepiece,move the
ruler so that one of the millimeter lines is just
touching the left side of the field of view. You
ruler will look like the diagrambut you will be
able to seemore millimeter lines sinceyou are
working at a lower magnification.
To determinethe diameterof the field of view for
the scanninglens,count the number if millimeter
lines (actuallyspaces)that are visible. You will
need to estimatethe diameterto the nearesttenth
of a millimeter.

Field of view

2 millimeters

Since the micrometer (pm) is the preferred unit of measurementfor use with the microscope,convert your
measurementin millimetersto micrometersby moving the decimalthreeplacesto the right (effectively
multiplying by 1 000).
Scanninglens (4x, 40x total) field of view:
mm or

Next rotatethenosepiece
to the low-powerobjective(10x, 100x total) anddeterminethe field of view. Count
the numberof millimeterlinesthat arevisible. Sincethemillimetermarksthemselves
arenow ratherlarge,you
will needto movethe edgeof oneof the marksto the left sideof the field of view andcountfrom the left edge
of onemarkto the left edgeof the next. Don't forgetto estimatethe nearest0.1 mm.
Low-powerobjective(10x, 100xtotal)field of view:

mm or

pm

Becauseunderhigh power(40x,400xtotal) thethicknessof oneof the millimeterlinestakesup practicallythe

entirefield of view, it is difficult to estimatethe diameterof the field veiw underhigh powermagnification.
The diameterunderhigh powercanbe calculatedon paper,however.Herearethe steps:
l. Divide the magnificationof the high-powerobjectiveby the magnificationof the low powerobjective.
Showyour mathin the spacebelow:

2. Then divide the diameter of the low-power field size of view in micrometersby the answerto the step
one above. Again, show your math:

High-powerobjective(40x,400totalfield of view:

mm or

pm

Sampleproblem: Hereis whatyou seein your scopeat 100x(usingthe 10xobjective):

Forlow power(10x,l00x total):
Edgeof ruler

Field of view

For highpower(40x,400xtotal):

Millimeterlines

2 millimeters

("

Recordyour group'sfield sizesfor the scanning(4x, 40x total),low-power(10x, l00x total),andcalculated

high-power(40x,400xtotal) on the datatablein class.The classwill adopta standardsetof field sizesfrom
classdata. Do not write anythingin the sectionbelowuntil all the classdatahasbeenrecorded.
Classstandards:

Preparea wet-mount of a human hair. Observeand draw the hair at scanning,low, and high powers.

Low-power

High-power

To calculatethe width of a humanhair, estimatehow manycouldbe placedside-by-sideacrossthe field of

view. Divide the sizeof the field of view by this number. Showyour mathfor eachproblemin the space
below:

/t

Practice problems:

HumanHair l00x

HumanHair 400x

Preparea slideof two crossedthreadsof differentcolors. View the threadsat low poweranddrawwhatyou see
in the spacebelow. You drawingsshouldincludecolor. Estimatethe width of the threads.

CrossedThreads
Magnification

Width of a thread

pm

\liJ U\ c,*1"^\q{, ons :

<.qrhPl"[" o\

Make a temporary wet mount of material from your'oPond inaJar" and view it using the microscope. Attempt
to chooseone microorganism and draw it in the spacebelow.

Pond in a Jar
Magnification

Width or length_
\^ri A+h

pm

c.o.tc\^lslions :

Continueyour work with the pondwaterby preparingadditionalslide(s)andlocatingat leasttwo other

microorganisms.Draw themin the spacesbelowandusethe field guideto identi$zthem.

Magnification
Width or length
Cq[o.rt\.^li snr ..

Magnification

pm

pm

Width or length
C"o.l.-,rlq{'on;

Label the parts of the microscopein the figure below:

Looking throughthe microscope,in what directiondoesthe letter "e" appearto move when you moved the slide

to therieht?

to the left?

awayfrom you?

towardvou?

What is the abovephenomenoncalled?

Calculatethe total magnificationof your microscopefor the following objectives. (Show your math)
Scanning
Low-power
High-power
What happensto the focus of the letter'oe" as you changefrom low-power to high-power magnification?

When the aboveoccurs,your microscopeis saidto be

How many times is the magnification increasedwhen you changefrom low-power to high-power
masnification?
What happensto the size of the field of view when you changefrom low-power magnification to high-power
maenification?
How many micrometersare in a millimeter?
Give the alternatename for the micrometer:

Giveits unit:

Give the unit for the micrometer:

)a