Using a Microscope.pdf

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Using a Microscope

EQUIPMENT AND MATERIALS

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BACKGROUND

snippetfirm c’m of tic’ Pork

Biology as a modern science would not exist without the microscope, just as modern

would not exist without the telescope. Both perform the same function: making the in

visible. Without microscopes, biologists would be literally blind to the vast majority of

around them.

When we took ooi fir‘-1hiph schoolbrolopycourses morethin

41) you a,po.they fomused_{on sum vey sepruents—sucir as}

ommmmmsmg mht tm-roottypes_{of pOuts for gross similarities and}

differences—cod dissectmons of frogs and other unfortunate

spocimmmeos Wwo me lucky to pet iiia fewminutes a week usmnp

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Thit wasn’t because sur’Jeys and dissectmons w

way_{to learn biology It was a matter of necessiti}

were very expensive, and most scimool budgets to provide enough of them. Many biology classrc only one microscope. Or none Nowadays, altho emexpensive microscopes are available, most sd

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it lii.I 1 iwO iicrosc.Ope.

it h, ita s tWeet wik he using the inic re cope every

cliocnt of every day, lint that the rnicrosco[v.r tots to be

lilalbeerr the spur of the moment Every tim you encounter

1hirg new to you your Prof thonoht Ocred wonder

ii firs looks lW urirt ufuruulicrt51OI)r it iiii Iiot,s Or’

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nIpossible yenWoolfdent cite a ivirk ra Lii n’l5cii

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i,eopevtafuiri furarrnkigiuirel I iv rrfr kuirt

tic (oven {aor lii. •iir;. fio desk (5riis, fi ; i

.me bed image of ,r ut vork ing oscrosciipebench We dicto’

nan up or move anyft snot before we shot this image, other thin removing the cover rein the mucroscope.)S wuh:hing from uuslng ihe computer to using the microscope t:kesliterallyseconds,

.‘ilsch means there, riveran excuse not to tuck snrrmeftiing

ifiiirder theiOicrns( rpm.

cr rutmnodnl it orIon, pes ui mimir’. roil ii

in different, rut nr,mnins lilIruricec hi.e.1 rrorrrirforhI to intrO. mbnt;inirnxi,t,ivi,nic.srLipiitirnrin c /,iiiniHi/

itStInter un the pro.nrco or ahcniuce’ t rn rn,rfuern

mt. which allows the nnin’reicope no hetilted amnif wfmrtfunrthe

,urrroscnoe focuise, iv muinvnn;g thestaureor the body tube

,ihrtever thepanticelar confguratiorm ofyour nnicroscope,

it’s irrmportant thatyou he able to identify eum.fr feature and

understandhowitisriced properly.With prictice, operrting

/0111microscope should become second nature, Forexample

you shouldn’t have to think about which direction to rotate the focsknob to open or ‘lose focus or which directiorr to turn the mechanical stage controlto movea specimentotherightin

/nuijrtielri of view.

rmfdition. tolearningto etc the microscope itself. enr ai.sn

inertto learn some batic nuucruscopy trrclmnrrqrres,immclui.tinrf

crcslide making, rn,mkirig whole moor its. sriirar irmuurrms.arid

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BUILD A DARKFIELD APPARATUS FROM POCKET CHANGE

So far in

this

_{lab session, we’ve used only bright field illumination, which is all most sti}

science enthusiasts ever use.

Dark field illumination

_{reveals structural details that are}

with bright field illumination, and is worth exploring. Fortunately, it’s quick and cheap

nearly any compound microscope to use basic darkfield illumination, at least at low tc

mag nit cation

Research-grade (very expensive) nricroscopes include special ilarktuc’ld corrdeiiser sand oblectivex. wrth lower thin standard roimer cal api’rtrires to optimize daristiild ir’nage quality. Although they arenot_{equipped with special darkfield} coridini-ersiii _{ol”iectivs,}sorTiE)stLident-g5lde nicroscopes include the hardwarr’ i’ieeded tousebasic; drrkfield ullrrrrrrrritiun,

or offerit_{as an inexpensive opton. You can also buy a darkfield}

assembly for $110cur so_{that tori be used with rn;rny standard}

microscopes._{but thor es really no need for}_it _{You can burld your}

Lab I-i Using a

own darktield apparatus in less than a second fc

(literally), and get resLilts about as good as yoLir $50 apparatus. Figure 1-1 6 shows you how to di

Yep. it’s thateasy. .cistplaceti_{coin in the center}

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510’ pOsits) r. cit t1C coo, ptace.110)00 o !o papa 50iii! •[e rtoat.o.r ,.i.orj l..rst t•he

1 hragrrr iisrnaI jarodoc’ owtrr (r.tc.nior.it

11.11 ‘‘l.’O.i’’ ii’,. ft’! Hi Ii1 iiO’L,’ “10 “‘ri rnr afro

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atioo

‘111111 irony huH. i.opes, thociirrrr tlrrf seiko irmtty vinll

with the 4X or lOX objecbve hut it you vaot to o’er darkficrd

ulluminahori with your 40X obtuctive ‘joint need to do a but nero work and sperunfafew Inure cents With a 40X objective,

the 1035k urust he be aii,nller thin a coin rod closer to the

rlraphraqm .ini toe boltorri of hi u:oiritin’rrrr. the titter Irolder 5

ii ohvioin’r toe. hen

‘i’ouii riceda piece of phrss or trio transparent pl,netic that fits

roteor ontop of your substage titter holder You can usean

or iirrary glassslide, ifittits betweenthetopof the filter hinder

ridthe bottom of the ,lianhr,n.nru a0)enrrbly. or a pioce otsOft,

li.nrrapinrent plastic rWe used .r plastic rnrcrnscope slirie, which

iiorbitsis,.rbent illplastic runiroscupo ‘,lndes aregoodfur)

(ijuil also riced an opaque, roundmask How big? We cant

..iy other than that it mustbe smaller in diameter than the

or i mumopeningof yourmicroscope daphragrn—probahly

mm Ii ‘in rlber ideally, whenif‘a focusedyouwantthe dark

oint:ral par iri5unji5 ih bripht rn!) to he iuinrt ‘tuirlntly t,)nrr.:;er

troth of an own.hairs ir40X n. htocIrae,w (iri.h rs p rcnh:rl...dy nbc

it mm Hnm’‘usia ri tiremna’okiii eden to r.urr:udoi.:o.iclark.iin.)

:0 it ri f’ .iii’.imi’t-5’’rf-,’ mnhiiC,i, .rt ur’l’’iit,

.1 /00‘dir by pt,ni.iHit the .liti’ iou on brtr”,tw

uifis the 4!( and lOX objectives uithe a Lige, Rot,..ntot.l nit)X

‘niyective intOpositron, focus on the hns’ht heldill age, and

-‘f

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oot 0 ‘e’ln,mnrhr r’’ri15)01510’ nil ,‘onfr”riser Iii u’,toi’,’j

o,.irs—i iii’nt_)f’t:r,ihfy t_uii.000it-slcri_’ioin’.nrf bt b-at!

rijrnmr,i,k -i/o

t withaunn,iok about itor in(.rn.lm.iarfor inr.;h) in

rfi,’rrr’for die usorf small stncky label lot, hut you cn’i also

O hree .0 pi...inn 11 1 [lii richOrit cir n”, urnI’. is- pijin rr 0 rink ai!,rihinnrif lirO tort-ntis to ‘rb k [Iso rn,r’,klottie

i__I ‘die or ,‘l,roati’ln’ -red positionj.tor theliOr’r twIrler

diitsbragrrr toits wstest settmrrg.Move themask,nrounrf orit:

mt,nrkr:erifralportiun,rppo,nrsirithefinlrfofviewriidthenclr

lliodiaptrr,rgnitoraif offatlhrnt,rthrn,iirrruilusarrrrrnrf their’ it limed irk j.r’t ‘fii.”i 1 fo’ier hrm’ofirr’ l,’lrlof‘imirw yr.iiltIn

01,1 k If lip ‘l,rrk corrtr,rl -pr.)l a ninth IliCer

of a n.y ‘jar mlni’prl ‘trrr.ntler nrm.rax iili’Hiy ‘j’u’,a.nrrn

io.oak10hrijduce I,.ru k ceinin spot [hr.itS ji..rst‘;Irnrhttyi,ii

th,,nrthe field ofvIew,

ifcourse prorfricmnrg a reasonably circular rnakof ‘ierrhi

ln,inni’ter :111her,litfmcnrlt Wetoed rnsrrrg 1 perrn’anrerrf

rri,ukertoprn:dnicn tinyblackerrcles, trilltirose ‘irHles wore

neiltierriert’rctlyroi.nrnd near comnrpletelyopanirlo. Ehmnkrmrg

,ihurnt it,werbecirfeiltotry usingsmalldrops of opaquepain rpplied with atoothpick, which worked perfectly after a bit

if expermnnerrtatrunr taproduce opaque crrcles of mysize w

-air trod.

thisfirst firingyou’ll rrr.rtrce i.nsing danktielri i1iunflnln,rtiOo at 1

nmnrgnntrcatnon is thatthe irn,nge is very dim fhat’o unavoirl,

because the only light err the specimen is coming from the poripher’j Youcan rrnprove flatters somewhat try removir!

trourfd glass slurnirrator m’lnfluartr disk

1)11cc you have a mask of theproper size,keep it stored sal

‘0 that yr.nu don’t loch to n mnivent the maskeach time you to n_Iso darkfnelcl ullumrnatnonat 400X.Unfortunately, rt’s ire

possible to build a field -expedient darkfieldapparatus for

‘auth fheIOOXoihonnrnersion obtective. Or, if itns, ‘ve can’t I

illit how todoit.

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Using a Microscope.pdf