How to lmprove PhotographyThrough the Microscope

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How to get the most out of this bookletThis booklel ls div ded into four ma n sections. ltems marked with a a I st the min mum informa-tion that newcomers to the art of photomicrography need to know, whie llems left unmarkedare addressed to users who have already taken photos with the mlcroscope but are not satisfiedwith the results. By relerring to sections 1 to 3 for picture taking and section 4 for checking,you will find this bfochure a reliable help the next time you want to take a photomicrograph.

From pEparation to observationlvlicroscopes suitable tor photomicrography . . . . . . . . . . . .8Equipment suitable tor photomicrography. . . . . . . . . . . . . .9Various types ot photomicrographic equipment

a n d p e r f o r m a n c e . . . . . . . . . . . 1 0aEquipment needed for photomicrography .. . . . . . . . . . . . 11

Suitable localions for sett ing up thie microscope ... . . . .12aObjectives and photo eyepieces suitable for

p h o t o m i c r o O r a p h y . . . . . . . . . . 1 3aDifferences in the peripheral images depending on

ob iec t i ve des ion . . . . . . . . 14aDiffeiences in re-solution depending on lne lype

of objective . . . . . . . . . . . . . . . . 15asome hints about the objective. . . . . . . . . . . . . . . . . . 16- 18

Selection of a condenser. . . . . . . . . . . . . . . . . . . . . . . . . . . 19a o b s e r v a t i o n p r o c e d u r e s . . . . . . . . . . . . . . . . . . . . . . . . m - 2 1a H a n d l i n g o f s p e c i m e n s . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2 2

Use of the f ield ir is diaphragm . . . . . . . . . . . . . . . . . . . . . .23lJse of the aperture iris diaphragm and its effects . .24 - 25

aBasic focusing methods . . . . . . . . . . . . . . . . . . . . . . . .6 - 27How to spot dirt and specks of dust in the optical

systems of the microscope and the Ohotomicro-g r a p h i c a l t a c h m e n t . . . . . . . . . 2 8

aHow to clean the microscope frame . . . . . . . . . . . . . . . . . 29a H o w t o c l e a n t h e o p l i c a l s v s t e m . . . . . . . . . . . . . . . . . 3 0 - 3 laHow to c lean an o i i . immers ion ob jec t i ve . . . . . . . . . . . . . . 32aHow to clean specimens . . . . . . . . . . . . . . . . . . . . . . . . . . .33

How to obtain good printsHow to obtain good color prints . . . . . . . . . . . . . . . . .60 - 61

aPreparing black-and.white p nts . . . . . . . . . . . . . . . . . . . . . 62N l a r k s o n t h e p h o t o . . . . . . . . . . . 6 3How lo avo id marks du r ing deve lopmen t . . . . . . . . . . 64 -65

SectionlFrom pEpaaation toobseryation

Sectlon2When laking pictur€sin photomicrography

SectionSHow to obtaingood pdnls

In the case of poor photos

$ection4Trcuble€hooting

tr:oJclion ol eouiDmeni suilable lorGE-Jal on and phoiomicrcgraphy, basiclti.€dge, observaiion procedures,

When taking pictures in photomicrographyBasic inlomalion in photomicrography

Operating instructions for models P[,4-1oAD andP[,4 IoADS (with 35mm camera back) . . . . . . . . . . . 36 - 37

M a g n i f i c a t i o n o f p h o t o g r a p h i c e q u i p m e n i . . . . . . . . . . . . . 3 8Differences in resolution according to the combinaiions

o f o b i e c t i v e s a n d e y e p i e c e s . . . . . . . . . . . . . . . . . . . . . . 3 8Setting ot photographic magnification (effective

m a g n i f i c a t i o n ) . . . . . . . . . . . . . . . . . . . . . 3 9F r a m i n g o f t h e s p e c i m e n . . . . . . . . . . . . - . . . . . . . . . . . . . 3 9Check ing o f t he f i n i shed pho tomic rog raphs . . . . . . . . 40 -41

Pholomicrography techniquesE x p o s u r e a d j u s t m e n t l e c h n i q u e s . . . . . . . . . . . . . . . . 4 2 - 4 3U s e o f t h e A E l o c k . . . . . . . . . . 4 4 - 4 5Compensa t ing fo ra f i lm ' s rec ip roc i t y f a i l u re . . . . . . . . . . . 46

color photomicrographyO C o l o r f i l m . . . . . . . . . . . 4 1O T v D e s o f f i l t e r s . . . . . . . . . . . . . 4 86 Differences in color rendit ion depending on the

combination of t i lm and l ight source . . . . . . . . . . . . . . . 49E Differences in color reproduclion depending

on differences in color temperature . . . . . . . . . . . . . . . . 50a Dif lerences in color rendit ion depending on lhe lype

o f f i l m . . . . . . . . . . . . . . . . . . . . . 5 ' 1a P u r c h a s e o f c o l o r f i l m . . . . . . . . . . . . . . . . . . 5 2

Use of color-compensating (cc) filters when takingc o l o r p h o l o s . . . . . - . . . . . . . . . 5 3

a T e s l p h o t o g r a p h y . . . . . . . . . . . . . 5 4Black-and.white phologEphy

a Black'and-while tilrn .

4 F i | t e r s . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . 56

555657

cE rc methods, etc. are explained in.esylsundeGland manner.

ranging linished photomicrcgnphsCng to their characteristics you can

:-€ causes for iailure and take cor-

aComparison of ditterent film brands

Pho tog raphy w i th Po la ro id f i lm . . . . . . . . . . . 58Trouble-shooting

Problems in tinished photos and thok co.reclionPoor color reproduction . . . . . . . . . . . . . . . . . . . . . . . .68-71Blurred image . . . . . . . . . . . . . . . . . . . . .72-73T h e i m a g e i s i n l o c u s b u t n o t s h a r p . . . . . . . . . . . . . . 7 4 ' 7 9Oojecls olher rha" the specimen imaqe appeared

o n t h e i i l m . . . . . . . . . . . . . . . . 8 0 - 8 1lJneven brightness . . . . . . . . . . . 82 - 83

l*, t"*'** and exprafanon orl-r"= p'ocedures needed fof pholGlt<r, drfferences in films and purchase

e action. In many instances, theshoollng seclion can also serve

+ct processing can ruin lhe best' This seciion deals wilh film denl, how lo order color Prlnls, elc.

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SectionlFrom pr€paralion to obseryation

lvl icroscopes suitable for photomicrography .. ... .. .. .. .8Equipment suitable fo. photomicrography.. ... .. .. .. .. .9Various types of photomicrographic equipment

a n d o e r f o r m a n c e . . . . . . . . . . . 1 0aEquipment needed for photomicrography . . . . . . . . . . . . . 1 'l

Suitable locations for setting up the microscope . . . . . . .12aobjectives and photo eyep;eces suitable for

p h o t o m l c r o g r a p h y . . . . . . . . . . 1 3aDiff€rences in the peripheral lmages depending on

o b j e c t i v e d e s i g n . . . . . . . . . . . . 1 4aDifferences in resolution depending on the type

of obiective . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15asome h in ts about the ob iec t ive . . . . . . . . . . . . . . . . . . 16- 18

S e l e c t i o n o f a c o n d e n s e r . . . . . . . . . . . . . . - . . . . . . . . . . . . 1 9aobserva t ion procedures . . . . . . . . . . . . . . . . . . . . . . . .n -21a H a n d l i n g o f s p e c i m e n s . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2

Use of the fiefd iris diaphragm . . . . . . . . . . . . . . . . . . . . . .23use of the aperture iris diaphragm and its eftects ..24-25

a Baslc focusing methods . . . . . . . . . . . . . . . . .... .. ..26-27How to spot dirt and specks of dust in the optical

systems of the rnicroscope and the photomicrcgraph ic a t tachment , , , . . . . . .m

aHow to clean the mlcroscope frame . . . . . . . . . . . . . . . . .29aHow to clean the optical syslem . . . . . . . . . . . . . . . . . 30 - 31aHow to clean an oifimmersion objective . . . . . . . . . . . . . .32aHow to clean specimens . . . . . . . . . . . . . . . . . . . . . . . . . . .33

M icroscopes suitable for photomicrography1. The photolub€ accepts the pholomicrc

graphic camera attachmenl.

2. -Ihe

image can be focusedthrough the binocul lube.

3. The desired lighl inlensily matchingtre specimen conditions c€n be setby the odical palh seleclor,

4. Objectives should have high resolutionand good llatness (iat image all the wayto lhe periphery ol lhe Msual fielo.

5. The slage is rolalable.

6. The condenser is equipped wilh anaperlure i s diaphragm and cemeiingdevice lor aligning 10 the oplical a.xis.

z$""[if;ffffi;'lXll""*'ipd-r'h ------8. Sufiicient lighl intensity is assured.

Light inlensily is adjuslable 10 match theobservalion and pholographing condi-lions. lJnilorm illumination from low10 high magnific€tion.

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9. The microscope sland is resislantagain$ enemal vibralions.

o10. Provision lor insenion of filters

'The phoio shoE m.del BHS

Equipment suitable for photomicrography

1 . The c€rnea attachmenl accepls 35rnmor large{omat films. The 35mm camsraback is equipped with automatic fllm

2. The oplical palh can be changed lomatch photographic condilio.rs.

3. The measuring arca c€n be changedto match the specimen, (lntegraledmetefing 3oor-sfDt meiering 1%)

4. The camera attachment is fmlyclamped on the microscope,

5. Device for measuring color temperalure.

6. Wide range for setting lso/AsA sensilivity

7. Coanpensation for reciprociy law failureis carried out automaljcally for long

8. E\po€ure adjuslment, matchingsp€cimen conditions, is possible.

9. Manual exposure is possible.

10. Special applicalions. such as 16mmcine and 35mm lime lap6e pholography,can be performed.(A special corfrcl unil is reqrifed in

1 1 . An exposure lock mechanism {AE lock)is built'in.I'-

o'rne phoro shm mod€ PM.1oAos.

Various types of photomicrographicequipment and performance

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' Concernlng the ioclsing screen iorolympus oM system cameras:TTre screen can be changed to sul dflerenluses (OM.] OM 2. Olv-3. OM 4) For pholomcrography lse screen No 1 12

' Sh|]lter b !r may occuf li magnificaton sincreased When usng 40X and 100Xobleclives. adlust lhe light inlensty so llratlhe shltter speed is 1-2 sec. In this case,some types of coor fim may requne coor

Types ol pholomicrographic oquipment

Pi|nOADS/ OM s€des SLR came6

E se ollocuslng

The',ocus ng screen mLrsl be re-placed ior pholomicrography'

Photographicquallty

Shutler b ur may resull when 40Xand 1mX objecl ves are used'

Ease olop€ralion

.Manual winding oi35mm lim

.Use of cable fe ease

.Manual wind ng of filrn

.Use of cable release

.Use of Varmagni Fnder lofrocusng

Use ot exposure meter(EMM,7) .tlse ol buill-n exposure meier

Use ol largeforfial lilrn

: st check the charl to see if lhe equipment-:.uned ior pholomicrography has been com:'.iely assembld. fihe lypes listed below are-€ most basic attachmenls needed for takingroto6 ol stained specimens in lransmitt€d lighl.

. q u i p49!! !99!ie! !!!!9pgEP!r :ocusiig magnifier (mI :ocusing Telescope! 35mm camera back. iLrlomalic aeosure body

. :onneciing cordt lrgeformal camera back| :dapter for largeJormal camera, tulomatic exposlre control lnil

B _slruclion manlals

t :crm lor recording datarr leaning utensilsi blower, cleaning iquid,

?1S Ceaning lissle

Equipment needed for photomicrography

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Suitable locations forsetting up the microscope

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Ressons for unsuitable localions and corective measures

Unsultabl€ room Cons€qugnces T|Ealm6nl

.Localed too close to mechanical appliarices ormachinery that can cause exlernal Mbralions

.P aces in which the vibral on ol personswalking pasl can be transmitled

Blurred image as a rcsull of vibralions

.Remove lhe microscope Jrom the soufceof vibralions

.Use a slurdy lable as suppod

.Use a vibralion prool lable

.Use of lhe microscope near a windo /

.Place where room lighl enters the eyepiece

Brlghl light lrom the wndow prevents correcilocusrng

photoljght or flares are rcproduced on the

.Sei up lhe microscop€ near a wall

.Position lhe microscope in such a waylhat ihe overhead light ialls in slightty infroni oi the microscope.

.Cover ihe eyepieces with caps

.Shui oul siray lighl getling nto the eyepieceor the focusing lelescope by changing lheoplica Palh seleclor

.A dusry and dlrly room

.Place nea. a winclow where dust can enierfrom lhe oulside

B ack spols are reproduced on lhe specimenimage

.sel lhe microscope up in another room

.Cover the whole microscope with a dusf

Example ol a suitable room lor photomlcrography

Objectives and photo eyepieces suitablefor photomicrography

Cor€ct comblnalions ol obioctiYe and ey€pl€c€Obieciives:.' pholomicrography high'resolulion cbjec-la€s wilh flahess all the way 10 the peripheryr - visual field are requircd. Oi the LB (ong-:a"e ) objectives, seies S Plan Apochromal,a >an Achromai, and D Plan Achromal,._c oi lhe short-barrel objeclives, Plan,rccchromal and Plan Achromal types are

tlpto eyepleces-e pholo eyepiece is optically coanpensatedE .ermil the objective lo deliver ils fullr€.'cmance on the film plane. ll musl be:..'eclly malched wilh the objectives.

Combination with NFK Combination with FK/P

s Plan Apochromat eeri€s

Shod.bargl Achromal s€ties

Differences in the peripheral images

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depending on objective designAccord ng lo lhe lype 01 objectve the perphery oi both lhe observed imaqe and thepholographed rnage rnay appear oul ol locusThis efiecl s caused by lhe pe.formancecharaclerslics oj Achromal lype oblecivesUs ng Pan Achromal oblectves however.wi res! t . a sharp and Tlat mage exlendnga lhe way 10 the perphery of the Jeld

rhe rierd ,rarness or a D Pan Achromai objeclive eiiend nq a th€ way lo the pefprrery s superof torhe one orovided bv a D ach/omal 0b _.ci va

Difference in resolution dependingon the type of objective

:- .: -: on n lhe center of the lield oi borh_- :cserved and the pholoqraphed images: _. : according to lhe type of objectve._ - -:c cass objeclive series, S Pan Apo-

:-:1s. as wel the S Pan Ach.omal- :: crovide superor resolulion.

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: ; i- Apochromat 40x. NFK 33XLD ligss-ffiJIU*H

Gives srrarp ,esorutor .r lh€ whoe mase. allowino obseflation oi mlnul€ dela s

Some hints about the objectiveIt ls importanl lo choos€ en obi€ctiyesultable ior your sp€cltlc purpo66For all objectives, proper use depends on lhespecilic purpose, some types requiring someadjuslments. ln order to tulv rJtiiize theobjective, you should know the meaning oithe various numbers and letiers engravedon lhe objeclive barrel.

objFljw typ€ and hagntficationNum€icar apenu,e (N.4.)

1. Obi€ctlve wlth con€ction collsrIn objeclives with a numeical apenure (N.A.)above 0.6- €xcluding oifimmersion l}pes-lhe thickness oi the cover glass slronglyailecls ihe image qualily. Cover glass lhick-ness is theoretically designaled al 0.l7mm,although in actual praciice this may vary by+0.3rnm- By oplically compensaling ior thisthickness devation, the correction collarassures the best image.

Method ot adiustnent2. Obieclhr€ wlth lds dlaphragmSome obiectives abo\€,lOX are equipped withan iris diaphragm. This diaphragm prevenlsdirecl light frorn enteing the objective inclarKield or lransmitted light lluorescenceobservalion. By watching both conlrast andresolution of the image, lhe apenure canbe adjusled to the optimum posilion.

(1)Set the scale lo lhe 0.17mm position,

(2)Rotate ihe coreclion collar 2 or 3 gradua.llon marks (0.02{.o3mm) in the directionoi 2, and refocus. lf lhe image is sharperthan under (1), rotate the collar anolher2 or 3 graduation maks in the samedireclion and tocus again.

(3)lt you are not satisfied wilh the imagequality, try rotating the collar in the oppositedirection tor 1-2 gradualion marks, r+focusing and comparing the image.

(4)Gradually reduce the amount oi rolalion otthe correclion collar, and try to iind the opiimum condilion by repealinq steps (2H3).

r-Ihe abo,/e adjustmenl procedures muslbe repeated every iime lhe specimenis changed.

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D Pran f{X, S Plan ADo 100X, Nc s Pt.n ADo 100x

: \o-cov€r (NC) objeclive::cmens wilhoui cover glass such as

:-: objectves bearinq the mark NC: :over glass attects the imase c arity,

.!h ch is padicu arly obv ous in lhe case:.tves with a large numer ca aperlure.':s!1, when observlng spec mens nol: :C by a cover glass at hqh maqnl

: specia NC obleclive is used.. ewing ufcovered specmens thfough

: .bjecllves, on y poor images w th'ares and insuficieni resolulon can

-r s malk indicares rhat no cov€r

-ous LB s€ries no cov€f objecl vesi: rn 40X, NC D P.n FL 60x NC S Pran: : ! a^d NC S Pan rmx dry

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)'r 'l /;l- (sp@imen w ihoui coverolass NcsPanApoloox

4. Nocovor lmx dry objecliveHigh.maqn l calion I00X obleclives are uslalyoj lhe oll mmersion type bul ior the nocoverl00X objecl ve a dry type is ava lab e Usingthis objective logelher wth other non.imme6on objeclves, e.g. nGcovef 40Xand 60X. obviates lhe need lo pul mmersiono on llre spec men slide The l00X dry typelens can aso be used for pholom c fography.bLrl ior achevng oplimum piclure qua ily,the lse oi an oil'immerson objeclve s

SDecimen wlhoul cove, s ass S Plan Apo r00X

Numdi€r aperluc (N.A-)Mehanial llb€ l€nglh

ofthenlcD6cop€

Obletlve lype and m.gnific.tlon

Tnlckns ot lie cdor Olas

Oiljmmersion objeclives have a numericalapenure above l-0 and use manulacturer-speciiied oi belween the lront lens ol theobjeclive and lhe specimen. (Oil mmersiofobjeciives carry lhe mark "oii .) In order iomake full lse ol the resolving power of theobjeclive, il is preterable to use oil alsobetween lhe condensef front lens and the

(2)Rolale lhe revolving nosepiece so lhal lheoilimmersion objective is pointing towardsyou, and apply oii io the iront lens ol

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'Use only oll specilied by themanutacluretli you us€ old Cargill oil or cedar oll, lheobieclive cannot display ils lull potentiat,since their difiaclion coetficienls differ iromlhe nominalvalue. li the oil is llnted. it affeclslhe qually of color rcndilion in color photos.Theretore, il is advisable to use only manufac.lurerspecified oil. In parlicular for fluorescenceexaminalion, use only lhe iluorescenceireeoil provided wilh the lluorescence microscopes.

clicks inlo the lighi palh. Make sure lhailhe objective lronl lens is tully immersednto the oil on lhe speclmen slide.

(4)lf the irnage is not in vieq slowy rolale lheline tocusing knob lill it comes into focus.

I Make sure that the objective fronl lens doesnot gel 1oo close lo the specimen, sincelhe image will deteriorale il air bubbtes gelinlo lhe oil. ll a haze seems to cover parlsof lhe image, even lhough it is in focus,swng the nosepiece 1 or 2 times frorn lhecljck slop in order to remove be air bubbles.

5. Oil.immersion objectlv€Use of the oil-innercion obiective

How to apply the oil(l)Focus on lhe specimen, wilh the 10X,

or 40X obiective and bring lhe desiredspecimen detail in ihe lietd of view

Il } tall- r lrbal

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(3)Apply oi lo the specimen surface androtate lhe nosepiece untillhe oii.immersiorlobjective, ils tip likewise covercd wilh oil,

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lf the lmage still does nol improve, removethe eyepiece as illuslraled above, checkior air bubbles by viewing the back tensof lhe objective, wip€ oii the oil andreapply oil.

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Nlmsdcal aperllB {N.AJMehanical lub. l.ngth

ol lh. mlcrosope

ObjeclivB typ. .nd magnltrcatlon

Thickn6s ol th€ cd.r glss

(2)Rotale the revoving nosepiece so thal lheoil-immercion objeclive is pointing towaldsyou, and app y o I lo lhe lronl lens oflhe objeclive.

'Us€ only oil sp€cified by th6manulaciulerll you use old Cargill oil or cedar oil, theobjeclive cannol display ils fu I polential,since lheir diffraciion coeflicienls ditfer lrcmthe nominal value. lf lhe oil s tinled, ii aiteclsthe quality of color rendition n color pholos.Thereiore, il s acfuisable to use only man!1ac.turer-specifed oil, In padcular for lluorescenceexamination, use only the fluorescenceireeoil provicled wilh the fluorescence micrGcopes.

clicks inlo the light palh. Make sure lhailhe objective lronl ens is fully immercedlnto the oil on lhe specirnen slide.

(4)lf the irnage is not in vieq slowy rctate lheline locusing knob lll it comes into focus.

I Make surc thal the objective fronl tens doesnot gel too close lo the specimen, sincethe image will deteriorale il air bubbtes getinlo the oil. lf a haze seems to cover parlsof lhe image, even though it is in focus,siwing the nosepiece 1 or 2 limes l.om lheclick slop in order to remove the air bubbtes.

5. Oiljmm€r$on objeclhr€Use of the oil-imnercion obiective

How to apply the oll(l)Focus on the sp€cimen, wilh the 1OX,

or 40X oblective and bing the desiredspecimen delail in the field ol view

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Oijmmerslon objeclives have a numericalaperlure above 1.0 and use manuiaclurer,specilied oil belween the iront lens oi theobjective and the specimen. (Ollimmersionobjectives carry the mark 'o l ,) In order tomake lull lse oi th€ resolving power ol theobjeciive, I is preferable io use oil alsobelween lhe condenser lront lens and lhe ob.,

(3)Apply oil 10 the specimen surface androlate the nosepiece untillhe oiljmrnersonobjective, ils lip likewise co\€red wilh oi,

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li lhe image slill does not improve, removelhe eyepiece as illlstrated above, checkfor air blbb es by viewing lhe back lensol the objeclive, wipe off the oil andreapply oil.

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:rrpcse ol lhe condenser s to eficenlly'le ighl emanal ng lrom the lighl

! on lhe specjnren. 1rr creale ightn!i :.is malch ng the obleclive and lhlsr. de a be11er image Depend ng on the

: .a abe. Padculary wi lh Lr l r ra ow mag.n obleclves such as lX 2X and 4X.

:ns ike uneven i uminalon and insul

: . r2aAc

: -2 SC

: - 2 C A

:-2 ULC

Selection of a condenserf icent amo!nl of rghl a1lhe perphery arekey 1o occlr Therelore make slre lo lse

lhese objeclles n combnal on wtir lhe! tra ow magnificaton condenser

n orde. 1r] obla n better photos wtir theS Pan FL 2X objectve. lse oi tire Lr tra ownragnilicalion condenser BH2 ULC s

:nbination of BH2 series condensers and LA obiectives

Condenser

Observation proceduresNow we are Ina y gettng to observalionbul firsl make sure that no dust or d ri son ihe obleclive, eyepiece, and specimen.lvake t a habt lo check ior dirt before yoLlLrse the mcroscope since d rt prevenlslocusing and resulls n poor image qua ily

1 Placement ol filters

Turf on lhe ma n swtch and adlusl thevoilage lo poston Pholo (ca 9V)

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(1)Pace lhe ight baancing f i l ler (LBD.2N)of the lighl ext wlndow.

t Adiustment ofr4 dislance

r,- ob .

(3)Place the specimen on the stage and focuswth a lox objective

(4)Adl!sl lhe nterplpilary dslance |]nl boU.leil and righl vew fieds mefge nio one.

interyupillary I 3

a

C

I

(2)Pace an ND i ter in the slot close 1o lheamp housng Dependlng on the objectver.agniiication and the densly oi thespecmen. Lrse lhis fltef so thal t providesenough brghlness for easy exa..inaton.

h^=-1.

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Diopter adlusimentrc Jsl lhe diopler lo sul lhe observels-,.esighl. Ihe melhod d flers according 10

I rnless the diopier is adjusted, parfocalily.r I nol be maintained ' hen lhe objeclive

: r;_en using WHK lox eyepieces, the.,:-s is adjusled with lhe focus ng knobs.. e observing lhrcugh the .ighl eyepEce^-€ aiopler adjuslmeni ing 6) on the letl'd): s then adjusted lor maximum image;'f lor the left eye.

(7)Al rhat time, adjusl lhe irnage so lhal thecross ines n lhe center of lhe lrame maskare cleary dslinguished as two separaieines. Then adjusl the focus by roiatingthe linefocuslng knob so that the crossInes and lhe specimen image are in Tocuss mullaneously. Alter compleiing I ghl_eyeadlustmeni, also adjusl lhe diopter lor lhelefl eye by rolating secllon A as n (5).

{ Condenser centering

(8)Alter locuslng on lhe specimen with lheI 0X objective, rotate the lie d slop A inlhe d rection of lhe arrow and reduce thefield irisdiaphragm d ameter io a minimum.Then slowly move the condenser lrom lopto bonom by using the condenser helghladjuslnrenl knob and stop at a posilionwhere the rield iis diaph€gm ir.ageis sharply deiined.

I Move the field iris diaphragm mage 1o lhecenier of the vsual lied wilh lhe con'denser cenlerlng knobs c .

@@a(g)Open the lield iis diaphragm image lntil

it almosl louches the periphery ol the vislaliield, and make some linal cenleringadjuslmenls. For normal observationconditions, make the diaphlagm slighlylarger lhan lhe visual field.

I lfthe field iris diaphragm cannot be sharplyfocused, check the thickness of the spec|men slide, Use slides wilh a lhicknessbelween 0.9 and 1.2mm.

III

-aa A'- us ng a iinder eyepiece tor pholomicrG

t=.y adjuslment is made by rotaling-: :r B. of lhe eyepiece al the right eye.

I S nce the \,pe of the linder eyepiece ditlersaccoding lo lhe size oJ lhe film used,choose lhe type suitable for yolr patticular

EwlrKloxE wFrhlox

E wHkiox41/4x31/4" Po aro ido

@wNrro {

Handling of specimensMake I a habLl toc ean tire specmen reg! arybolh before and afler obsetualion Jlsl aswlh lenses 1is mosl mportanl lo work wilha c ean spec men Nlake sure thal no dlslpairces stck lo the specimen when youstore I and do not toLrch the gass slrlaces

Ho ld l heA lass s l i dea l l he co rne ron y , o rhod l h€ g ass s l i d€ be lween you r r i n0e . s

, / t , i .F inoe@an lson rhe .ove r g ass The imaae i s bu i i ed aons t he l r ac€so l you .

I n9elpnnls

Notes on specimen placement on ihe stagewhen you pace a sp€cimen on the staqethe spec men irolder is opened wide and it s re eased rapdly. i1 wl h t the edge oj theslrde and damage the g ass. Aner coni rm nglhatlhe sp€cimen has be€nlirmly put n pacellre sprng loaded spec men holde. ..u$ beretracted sowLy so thal il genly lolches lhe

n case the s de is damaged carefu yrenrove the tny glass ffagmenls. I fragmentsare eil on lhe stage.lhey may calse inilres.or the specimen s de may be paced n at t€d poston on lhe stage. caus ng one sdeoi the v is la ie ld to be oLr i o i 'cc!s

one sde o i rhe v suar { ie rd i s b

Use of the field iris diaphragm. 'ed l r ls daphragm seves 1rr aolus l the- "ated area on the spec men dependn!'',e obleclve oowe. lh s daohragm paysr .a roe dur n9 Dhoion cro l raphy and i

._:. ode than necessary r uninal n! gh1. ecred ..d scaltered rre_o! a ly on the: ren eslLlrng in a oss oi mage conlrast: : ig down the l ed d lapnragm 10 l !s1:1d the irame rel c e area v/i resu 1 n

: rlreph c mages wih rmproved conlrasi:re ted rs dapl-rra_om s stopDe.j do!,/n: c ose io the irame retc e tire ohoto:!red ma_oe may be cLrt ai tfre corners! d aphragn,shoLrd ihereTore be ooen€dtrly more lhan 1l.e re:c e slro\ls

ljsing the Plvll0AD tunet mask locusing telescope (with 35mm Jilm)

' ' . . r - . " , , . . ' : ' : , 1 : i : i ; ' r

.l

Il.

I ' e rd , 3d aph fagm h3s been

Use of the aperture iris diaphragmand its effeits

The diaphragm rnounled on the condensers ca ed aperture rs diaphragm The iunclionoi this diaphragm is lo maintain opl mlmcondilions oi image resolulon. conlrasl andioca deplh by adlusling the nurrerical apenureof the lLrmnalion syslem depend ng on lhenur.erical aperlLre of the objective in use.For most specimens opllmur. irnage quajlys achreved il lhe apenure daplrragm sadjusted lo between 60% and 80% ol lhe

How lo adjusl the aperlure ids diaphragmThere are lwo methods oi adlushent:Pu oul lhe eyepece with lhe specmen nlocus and then adlusl lhe daphfagm !,vhllewatchng lhe rs al the rear ioca plane oithe oblecliv€ as n pholo (1): use the gradLraton marks on tire condenser. as in (2)

- 0)O r€cl viewng wilh lhe eyepi€ce

Reducino rh€ ap€r1ure nis da-prragm lo 6os0% ot lh€ numeicaapenure or lhe oblecl ve.

(2)Usnq lhe oraduar oi ma*s on rhe

Samp e Us nOa loxoblecrive wilha numercalap€duf€ ol0.25 and reducing ll to30%, ihaqGduar on malkon lhe connenser shouldbe ser a r 0 2 (=0.2s '03) .

s

Example Whsn uslng S Plan Apo 20)( NFK 2.5X

II -*o " *" "*n"* iris draphrasmI F€ge resolLilion deleriorales if lhe apertureI .: d aphraom is slopped down loo muchI r'h lhe exceplion ol specialy slained lhnI .ec rnens lhe diaphragm should not beI sDped doBn lower lhan lo 60% ol lheI nrencal apedLire ol Ihe obieclive.I I Ar effecl srmrl lo stoppng down theI rDenLrfe rris diaphragm can be achievedI :t moving lhe condenser clownward bulI :ris lends to interferewith lhe basic illumina'I :o1 iunclion of lhe condenser and resultsI _ uneven ilumination. Thus always use lheI : aphragm and clo not rnove lhe condenser.

I

$#ffiffiFully op€ned poslilon

&'/"Conn4l b enhanced. Details are alsocr€any vblble, .nd localdepth lsincr6ased, r€sulling in an oprimum

g./o

Re.olurion d.r.riorales as a resull

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Basic focusing methodsFocusing lor obseruationA lOX obleclve s used as lhe slandard forlocusng. thef the objecrlve s changed lromr(]X lo 4X and rrom rOX to 40X and i!(herlo 100X Do nolchange abrupilyirom iowmagn ficat on (2X and 4X) to h gh (40X and 100X)As a resLr t of lhe imled eye oi the obsetuerand the large ioca deplh oi low magnilicatanobjectves (2X and 4X) lhe ironl mounl l)1the h gh magniticat on objectve could touchthe specrrnen surtace when the revov ng

Focusing lor pholomicrographyFocus ng dlring photography s done etherthrough the locusing lelescope of the photographlc altachmenl or lhrolgh the eyeoiecesof the binocLr ar tube. When locusng throLrghlhe eyepieces. a jnder eyepiece mLrst beused Pror to pholomicrography the fndereyepece has 10 be foclsed by means oT i1sfocusng l ront lens. to make cear ly v isbedoLrb e cross lines as 1wo para el nes lnlfre cenler ot the lraming relice

1

- b

AJler iocus ng on the spec men $, th lOXobieclive sel the upper mt ol the coarseadllslrnent excurson willr the preioclsing

--)Fi-rl

l giII

Move the specmen detai 1{) be examlned tothe center of lhe vsua leld and increasemaqnilicaton by rolaling the nos€pece.

a' {

/n ErFinder eyepiece ol lhe binocular tubeFocusng through the binocLr ar tube spossbre wlh microscopes Vanor BHS. BFTand BtsTU

21

*r+

l - .

"i]". I Focusing lelescope ol lhe photographic

For pholoqraphy the specimen locus mLrslbe adllsled wth the same eye wilh wh chlle .ross ines were foclsed

I

l 31. Focusing when using 1x,4 and 4x

obieclivesFoclsing errors occ!r qu le irequenty wthlow nragn flcat on oblect ves Tierelore lsea locusng magnjer and adlu$ the iocLrsby iolow ng the procedures lsled beowBui beiore that the pholographer shoudadjust diopter a1 the cross nes oT theJocusng ieescope or the fnder eyepece

2. Focusing when using 10X and 20X

FocLrsrng rs achieved by adjustng lhecross Ines so lhat they are ceary v isb le.and then rolat ng tire lineJocusng knobLrnt both lhe cross nes and lhe mage01 the spec men are c leary v isbe s m!laneousy. By sLghly mov ng yolr eyesto a sdes. see i the posnLons oi lhecross ines and the spec men mage '1o nolshi l f t i r is s the case lhe Pc1!re sn focLrs t they do sh it. the tocls rs nots;

4r! l,^,*r f f )'rage is s ghlly oul oi iocls aner

. : o j magnlcal ion lse on y the f ne_j knob ior refocLrsing n partcLr ar

rse the coarselocusng knob for_! a h gh-magnlflcat on object ve. s nce. a danger lhal the obleclve w rLn- spec men ana uer oamageo

f s propery a gned and mls lw tl-r tire lne jocus n9 knob

3. Focusing when using ,lox and 10{x

Adl!$ the cross nes so tfral lrrey arec ear y v sib e. llren s owy adlust ihe locLrsot the speclmen image u71h lhe l nelocusng knob lnli cross Ines and lmageoi tl-re spec..en are c eary vsDle

: : 'eyeplece isorovdedwth a ocat n9: . n must be firmy nse(ed nto ther ' the eyepiece seeve f lhe p n ls

_-_t rnserted.correcliocLrsngbecomes:e and lhe magew beoLto i foc ls .

Camp the iocusing ma!nlfler on the'oclsing leescope. afd side the top secron n or olr thereby readluslLng the toclsal lhe cross lnes. FocLrs is co(ecl tbolh the cross lnes and lhe specmenmage are ceary vs ibe s imulaneousy

Basic focusing methodsFocusing lor observalionA lOX oblecl ve s used as lhe standard iorlocusng. then the objecllve s chanled fromrOX lo 4X and irom 10X lo.10X and ilnherlo I 00X Do noi change abrlpl y irom low magnirical0n (2X and lxl to hrgh {40X and t00X)As a res! t 01 llre lmled eye oi lhe observerand the arge ioca depth of ow magnificatonobleclives (2X and 4X] lhe Tronl mount oithe high magniiicalion obleclive coud louchlhe specimen sLrrlace when lhe revolv ng

Focusing lor pholomicrographyFoclsrng dlrng pholography s done eilherthrough the iocus ng leescope oi the pholo-graprr c attachmenl or tlrrouqh the eyepecesot the bnocuar lube When locusfg thro lqhlhe eyepeces. a fnder eyepece must beused Prior lo photom crography lhe indereyepece has 10 be focLrsed by means of lsfocls ng iront ens to rnake ctearty vrsbtedoube cross l ines as lwo parale lnes nlhe cenler o l the l raming re lce

3I

I,

, l

21

- ' ' b

After ioclsing on lhe specimen with l0Xoblectve. set the lpper mil oJ lhe coarseadjustmenl exclrson wth lhe preiocus n9

1-)E

Focusing t€lescope ol the photographic

For pholography the specimen focus mLrslbe adl ls led wlh the same eye wlh whichl"e .ross lnes were foclsed

fvlove lhe specr.fen dela to be exam ned 10lfre cenler o1 the vsLra fied and ncreasemaqn lcalion by rolal ng the nosepece

2 l al * -

Ir-'*r+

(

Finder eyepiece ol ihe binocular rubeFocusng lhrough the binoc! ar lube spossbre vilh microscopes Vanox. BHS. BNTard Br ITU

il1: : - ---

lr i

,

f s 1. Focusing when using 1X, 2X, and 4X

Focusng errors occur q! i le j requentywi lh

ow magn i cal on obieclives Theretote usea focLrs ng magnilier and adlu$ the ioclsby fo owing the procedlres sled beowBlt bejore thal llre pirolographer shouldadlusl dopler al the cross Lnes of thetocls ng leescope or lhe fnder eyepece

:,:,'"Hll"l -')I

Camp llre iocls ng magnifier on lheIocLrsng ieescope and s de the lop seclon n or o!1. ih€reby readjustng the tocLrsat lire cross lnes Focus s correcl 1both the cross nes and the spec menimage are cear ly v isbe smLr laneoLrs ly .

2. Focusing when using 10X and 20x

Focusng s ach eved by adlustng lhecross lnes so that they are c€ary vsbe.and lhen ro1aling the fne ioclsng knobunt borh the cross ines afd the mageot lhe speclmen are c eary vsble s imutaneousy By s ghlly moMng yoLrr eyesto a sides see I the posilbns or thecross lnes and llre specimen lmage do noishil li th s s lhe case the Pclure sn iocls li they do shjl. the iocus is nol

f s

: male is slghty oul oi jocus afterte 01 magnical lon, lse ony the lne: -! knob ior reiocusing In pancuar: lse the coarse locusng knob for

. rg a hgl r .magn fcaton oblectve. s nces a danger thal the oblecl&e lvr rln: spec men and get damaged

r: r eyep €ce s provicied w lh a localing_. pln musl be I rm y inserled into tl-re

::r y nserled. cotrect locls ng becomes: oe. andlhe magewi beol tof iocus

qt * -i - j

b - t

l fn -

propery a gn€d and musl be readlusledwth lhe fne ioc ls ing knab

3. Focusing when using 4OX and 10ox

Adilsi the cross nes so thal lhey arec ear y v s b e then slow y adlust lhe locLsol the specirnen image wlh the r nejocusng knob un1 cross lnes and imageof the spec men are c learY vsble

\

How to spot dirt and specks of dust in theoptical systems of the microscope andthe photomicrographic attachment

D ( and dLrst panc es are sor.el mes noteddur ng observaliof or phologrpahy. but pinponting the r exact localion nray be diiiicuitWhen photographing imponant Spec menswhich cannol be photographed again 11 canbe realy i rus l ra l jng i dust padices are vs ibeon tlre pclLrre. An eliective method s there-fore requ red thal w hep you deiecl

I S inced r la t ocatons no r_and i maycomeirom panicies oj lhe n m lisei (oadng. j mtranspo(). regular checks ard necessary

I lf the aperlLrre daphragm is.edlced below30%. dirl and dusl Specks becorne cearly

Dlst specks on the pr sm insde the obser'valion lube. on lhe inside of the objeciiveand on lhe jnsde oi the rn croscope basecannot be wp€d oJl eas y. iceanngbecomes necessary conlact your Oympls

\frf' oescrirrionObser.grapny Melhod ol verilication

Rernove the adapter ior large{orrnal camerasand check ior dirt by peerng ln lhrouglr the lopli you spot dilr unsc.ew and remove llre re ay ens

Rotale lhe lop lens eement as you observe lhe

:91 Camera prism a Sel to,Time mode, open lhe shulter and peer in

Either remove photo eyeplece and check ior duslq"lig 91 9l r".9"" phgto 9lep ece in prace, rotatet and check for movng dust parlc es.

-_

lhrough lhe foclsing leescope or lhe fndercha^ge rfe op_ical pa f 4- le alernarely ooqetu g

r Oplical path

Tube englhO correcton

Remove lhe observation tube irom the mcroscopea body and check lhe prisrn su.face ior fngerpr nts

or conlaminalionce ovF the obectNp roa l\e nos6prale d o- check t lor d rt or conlaminalon.O object ve a

_O Specmen

!0 Condenser

Observe lhe spec men and move lt in lhe fied.li dust is on lhe specmen, il wi also move.Remove lhe condenser lrom the mlcrcscope andlook for dlir and oi deposits.

^ . Check the iilef after removing it lrom the m.rG- ' scooe oase

Swilch on the lluminalion and examne lhe lens

8 Frosted gtass check lhe irosled g ass

; ; *

l? Lghl exil

j.i Co lecior lefs a a Aemove lhe amp houslng and check rhe ensesin lhe colleclor assembly.

z 199!li9

Bulb ^ - Bemove the bulb trom its sockel and check' ' sgns of backen ng fingerprnts. dirr etc.

How to clean the microscope frame:ii _s on the rnicro€cope lrame are lirsl..€d wlth a piece ol clolh weted with ai: amounl of neutral delergent, and then..€a clean wlth a piece of clolh lhal has)ir- ,mmerced ln luke-warm water But makeL-. roi io louch lhe lens section,rvhile:er_ ng the microscope ifame.\. : using organc solvenls which may--:ge paslic parls.

::r dusl conlam nation thal adheres 10= _ - pans and is difficult to remove, wiper: . piece of cloth or soll tissue paper that-3: :een soaked in a mixlure oi 7 parts etherr: I parls alcohol, Keep lhe m xture away+:- claslic pans lo prevenl damage.

Mslerials us€d lor th€ mlcroscooe

EEE

How to clean the optical system' p 6 o r a . 6 o o r . o t . r a n . p a . o d t . .

s . o . . r o e r . o r . o " - |surtace t can forma y be renroved wth abower Yo! shoLr d irowever. make a habloi covering the mcroscope wth a dlstproolcover after eacn lse

,- ll:e i\

2 l s

' i l ' f l ' n. s . b | i

To prevent scratches on coalings and oplical! ass remove din and dlsl lhat stcks 10the rsuriaces wth an ar gun or b ower brush

- -------"'- t

t..ii-'

' € ' "__

+'.I

Cleaning melhodT duslspolson optca glassessuch as lensesprisms and i ters are e11 unallended. thedLSr becomes d ficLri ro renrole and maycalse mold By always keepng optca glasss!rfaces clean. yo! avojd ma nlenance probems ard prolong lhe ie of yolr mcroscopeClean ng oj lhe ens suriaces app es ony 10exposed areas oi obleclives. eyepeces lirlersand condensers If lnlerna or malor cean ngbecomes necessary. pease conlact yoLrrO ympus N4icroscope dea er

Required lools

Wrap lhe lens lssLre aroLnd a wooden orbamboo slck as ustraled.

5-e

) t,

i A r gLrn or bower brlsh2 cean ng mxlure ot 7 par ls e lher and

3 pai(s acohol. or efs clean nq f! d3 Ol ps wood sl cka Sojl qauze. lens 1ss!e! Magn ryng glass An €yepiece can aso be

lsed n pace ot the magn f e

When clean ng large g ass slr{aces on bothsdes ot an accessory such as a j t€r fodtwo or three ayers of lens lssle soaked nthe cean ng mxture hod the accessory a lils edges and wipe lrom the cenier lowardsllre perphery as yo! s owly rotale I

When c ean ng tl-re suriaces oi the condens:and oJ the iighl exil gass hod a piece oilens lssue between yol r mdde and inderingers lod I afd $/rap i1 arolnd yoLr ind:ringer Then hod lhe lissue down wth yo!lhlmb wlrie vr'rpnq llre lens sudaces cea.

--v

T,* - ' l

wWhen cleaning a large lens surface, wipefrom the cenler lowards lhe periplrery if

i .,Qtt'

When cleaning a arge lens wilh lens tissuewrapped afound your llnger, yoLr should wipeIrom lhe center towards lhe periphery in acircuar rnolion. Aways, use a cean pornoriof the ens ilssue as yoLr rotaie your index

After cleanng, breathe ghlly on the enssurlace unli lhe whole surface has lurnedwhle, then check whelher ihe haze disappears un lormly. Spots where the hazed sappears on y slowly are not yet wiped

ti]- : ean ng, examne the lens slrface wlth. -.:t. [rng glass. ]i color rcfected iromt- -,_s surface ooks uneven, it is an ind.- :. :nat there are sl I dusl specks and

" . ewng ihrough the bonom oi an eye. ,oLr can use it as a magntying gass

tL, -=t \

. : , -YJ

:.:. lhe lens by pulilng a small amounl ol? : .eaning ilud or cleaning mi*ure on the. :'a lens tissue. Dlscard each ens tissue

How to clean an oil-immersion objectiveCl€an the oil-immersion obieclive dunng

Aner in sh ns obseryal on wth an oimmerson objectve. wel a pad oj cotton wooor a pece oi ens lissue wilh a sma amoLrntof cean ng mxlure conlanrng 7 pans etherand 3 pans acohol, to femov€ ol adher nglo the objectve S nce an oil 1 m w I oilenadhere 1o lhe obieclive fronl ens t sholldbe wiped clean l\/ice after eaclr JSe

llse only lactory recommended immercionoil and r€move it atter examinationi the o femains on the objectve ior a ongperod o i t rne. t wl l haden(eg cedaro)makng t diiiicLrll to remove even if yoLrwipe the ens repeated y Tlre lens may bedamaged ln lhe process Use ony speci€doi. and afler ]]se wlpe lhe o irom the mmefsion surlace of the objectve. keepng t

lj you lrequenty use an oll rlrme|son oblec-live olmay contaminate the sLrrjace oi adry t/pe oblecl ve *hen you change oblect vesTo prevent lhe o irom adherng to llreobleciive careiully rolale the nosepiece afterlower ng lhe slage. so thal the o does nol

I lhe mage of a dry lr gh-magnifica|onobleclve appears iuzzy. check for o thalnriglrt have adhefed 10 lhe 1ip oT the

6

w pe rhe llonl rens oi llre oblecl've r\1

Lower rhe slaoe slowly so lhal no oi llouch lhe rronl rens oi the dry objecr '

Alle. owerns lhe sla0e careru ry.olanosepiece and crick rh€ dry obleclNe

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How to clean specimensr,:ke il a habl lo cean each sp€cimen bolll:r'cre and afler observation. OlheMise, dirt-r dusi ihal you lailed to notice dlrngriervalon mght appear on llre pholo.::- cleaning the speclmen a son c oth, gauze,:, sece o1 lens lissue may be used wiihoul:.aing liqrid. Bll ii ihe conlaminal on s,-.Lrll lo clean, brealhe on the spec menri:re wip ng il. When ceaning lhe specimen,

*,nts to note during cleaning' iemoval of oi as well as roulne cleaning

:an b€ done morc easily il lhe specimen; removed lrom the stage.

: .ihen using ceaning mxturc or lens: eaning illid, use a moistened clolh or:?aniag lissue. Be cerlain not to apply:.aessive fluid, as il may seep underneath-. cover glass and darnage lhe specimen,

.b

--:re are Mo lypes oi sp€cimens: those wiltra : : .er gass andlhosewlhoulacovergass,rJ:- as blood smeaG cleanlnc sp€im€ns wilh cover qlass

+Cleanlng specimsns with cover glassos

?s when cleaning lefses, wpe ol1 lhe

-.s n a xylene bath. There are conta ners

:'3 dirl wilh a piece of lens lssue ighlly.'ened wilh cleaning mixlure. Because: cannol be complelely removed withroe repeal wiping unlillhe ol film

Cle6ning spcimens wilhout covertElss6s::-eing to uncovered speclmens cannola 3ed ofl. You can, however, remove lhe:. Tmelsng lhe specimen for 5 to 10

t/t:'::. horizonlal and vertica irnmerson,:..o€r selecllon of which depends on

cleanins spec mens wrthout cov€r orass

@

Section2When taking piclurcsin photomicrography

Exposure ad jus tment tschn iques . . . . . . . . . . . . , . , .42-43L,se ot the AE lock . . . . ., ., .44-45Compensating for a film's reclproclty failure.. . . . . . . . . .,16

Operatlno Instructlons for modds PM-1oAD andPM-1oADS (with 36mm camera back) . . . . . . . . . . .36-37

Magnification of photographlc equipment . . . . . . . . . . . . .38Dlfferences in re6olution according to the combinations

of objectives and ey€pieces . . . . . . . . . . . . . . , . . . . . . . 36Setting of photographic magnlllcation (effective

m a g n i f i c a t i o n ) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3 9Framiog of the specimen .... - . . . . . . . . . . . . . . . . . . . . . 39Checking ot the linished photomicrographs. . . . . . . .,O-41

Ocofor film , , , , . , . . . . . . . . . . . . . 47a T y p € o f f i l t e r s . . . . . . . . . . . . . . ' , | 8a Dlfferences In color rendition dgpendlng on the

comblnatlon of film and light source . .. .. . . . . . . . .. .,$a Dlfferences in color reproductlon depending

on differences in color temDerature .. . . . . .. . . . . . ., .50a Diff€rences in color r€ndition depending on the type

ot tilm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51O Purchas€ of color f i lm . . .. . . .. .. .- . . . . . . . .. . . . . ... . .52

[Jse of colorrompensating (Cc) filt€rs when takinocolor photos . . . . . . . . . . . . . . . 53

aTes t pho tog raphy . , . . . . . . . . . . . 54

a B lack€nd-wh i te f i lm . . . . . . . . . . .56acomoar ison o f d i f fe ren t f l lm brands . . . . . . . . . . . . . . . . .56a Filters . . . . . . . . . . . . . . . . . . . . . .57

P h o t o g r a p h y w i t h P o l a r o i d f i | m . . . . . . . . . . . . . . . . . . . . . . 5 8

tD

Basic intormation in photomicrography

1 Seting the lilm lormat 2 Load ihe film propody

Plsh lhe buton ior 35mm camem. Load the ii m nto the camera and make surerhat the end ol the iim does nol protrLrdebeyond the spoo groove OlheMSe tireJrame spacing may not be Lrn lorm.

7 Center the specimen to be, photographed in the visual lield R Fgcus and adjust the apeiure

v ms otapnragm

Operating Instructions for models PM-1OADand PM-10ADS (with 35mm camera back)

. r l3 confirm the winding or tne rirm

f 4

Wlnd the fim onlo rlre empty rake up spooand advance i1 10 irame m. I� To conflrmthat rhe fim s winding be cedain thal thef i m f e w n d n g c r a n k s f o l a t n g t l s n o t .rolale the crank 2 or 3 tmes n the d rectonoj lhe dotled arow lo pick up lhe slack olthe I m nsde the car.era. and aga n checklhe rewndng crank to coniirm the wndngof lhe i rn

V Adjust shullsr speed

Move lhe area ol llre specmen yo! wanl 10pholograph inlo the cenler of the visua ieldwilh lhe stage contros. When yoLr use modePM-1oADS. cenler rhe area 10 be pholo.graphed n the spot mete.ing seciion oT lhe

Focus on lhe spec men and adlusl thecondefrser apenure irls diaphragm so lhalsutabe cofirasl s achieved The aperllre.laphragm s ordinar y sel at 60% lo 80%or the objecuve nLrmerca apedure

Check lh€ shuner speed. Adlusl llre speedto between 001 and 05 seconds by usingND i itels

X

Color phologEPhySet lhe color lemperalure to malch the typeof color film used. Measure lhe temperatureat a blank area not covered t'v the specimen.

S€r lhe speed for lhe lih Lrsed.

aesure adjustment is based on the lolalion oi the specirnen.freler lo

Sel lhe characleristics ior reciprocity failurcfor lhe film used.

Focus is adjusted either through the tocuslngtelescope oi the PMI0AD/PM_I0ADS or thebinocular tube of the microscq*,(rcfer topages 2627)I The focusing telescope of lhe PM_1 oADS/

PM-1oAD includes different pholo lrameslor diflerent lypes of film.

I l}le lype oJ Jinder eyepiece \?tes wilhiilm size and each type of nnder elepieceinclucles several pholo frames indicatingdiile€nl magnilicalions oi NFK photo

The vollage posilion when not using the colortemperature meter cTF is:8.5-9.5V. for models BHS and4 or 6V lor models BHI and BHTU

Elackdnd'white photographySel the voltage above 6V Normally, a gleenlilter is used.(Fat enphaslzing padicular colors rclerta page 57 )

I!,De of fflmtldrl

helarclngffllor

Dlal pG!thn CIn

Daylbhi film t.BD-2N DTumsi.n lllm I-BT T

Press lhe shutler release butlon. Aftercompleling exposure, check ior the soundof ilm wnding.

qt

Magnification of the photographic equipment

errotogaptricequipment Photoevepiece FKNFK{ype P.type

Photo magnlcalon on the flm surface varieswth the prolecton lenglh oi lhe photographceqLr pment and lhe type of pholo eyeoi-ocelsed ln a cases pirolo rna_on fcaton scomputed by mutpyn! the objectve magn I cat on and the prrotoeyepiece magniTicat onvr th the coei ic ent s led in the tabe onrne flgnr

PM. lOADP]\,4.1OADS 0 5 x

P[.4 10M 0.5 x

PN,l,6

BH2 PM 6

09x

08x 0 4 x

Whef usnq models Pf.'ll0AD or PNrl]OADS rOX obleclive and 2.5X NFK pholo eyep€cePhoto man 1cal on = r0x 2.5 = 25

' . o

', 1

l.. ii ' ' o'

, ' l? ' ;" . ' '

? 1 ' t r ' - t l .

. , . . . 1 l t

1 r z l { . . t t . . .

. . " tr'

: ' ' t -

, 1 ' . ' ' .^ 1 , " - - - . ' " .

[ .

- t

, t .

Even tholglr lvhen lhe overa lpholo magn i calon s lhe same. reso Lr l0n !ares dependngon the comb nalon of obleclve and pholoeyepece The resoul on on the 1 m panes mproved j lhe magn flcaton oi the photoeyepece s ow and thal oi the objecl ve h glrAs the photo shows. a comb nalon of oblec.Uve 40X and NFK 2 5X provdes a c eardspay of even minLle mage dela Focadeplh. lrolrever. becomes slra ow

a rt

. e

I

aa

' t

. t t.,.a\ I

Differences in resolution according to thecombinations of objectives and eyepieces

Example: Pholo magnilicalion ol 100X

at'

Oble.l ve S Pan 20X and prrolo eyep ece NFK 5x

t ll a

tt

Obrecr ve s Pran 40X and pholo eyepece NFK 25r

Setting of photographic magnification(effective magnification)

n'en taking pctures, you have to Jirst decidear rhe desired enlarqemenl ralio. The mag.r_calion on ihe film pane is generaly:E:ermined by objective magnilicalion limes9oro eyepiece magniiicaiion. But the eliec

llve magnilicallon ol the piclure changes *The etfeclve magnilicalion is based on thewilh the nume cal apenure ol the objective, assumpllon thal ihe picturc is viewed wilhinand ls normally based on lhe lollowing re a- lhe closesl distance atlording dislincl vision,tionship, which must be laken into accolnl The values isted below do not apply lltenwhen enlarging lhe pholomicrcgraph. trying to proiecl 35mrn lormal slides.

5@N.4.<M<1qnN.A. N.A =nune,iczt a\dup at th. obta.iE M

Si6ctiYe.agnilication tx 't0x mx 40x 'tmx

tF olai.tclive

S Plan S Plan S Plan S Plan S Plan S Plan S Plan S Plan S Plan S Plan

t-4" 0.13 0.16 0.30 0.40 0.46 o.7D 0.70 0.95 '|.25 1.30

EHiveatnilication 65 - 130 80 - 160 150- 3m 200 -4m 230-Affi 350 - 700 350-700 475 - 950 625 - 1250650- r 300

Framing of the specimen-! pholography, lhe specimen ollen.rol lil inlo lhe ohoto frame. This oroblem

!e overcome by rotaling the camerain relalion lo the specimen, though

'nE

operation ot lhe microscope becomes morcd ficull. Herc, the iraming of the specimenis carried out the p€ferrcd way, by rotatingthe slage.

Phobrnicrography inuol!€s scientjfic pt|otcgradry, which makes it impolatn/e thal lhedDl€rapher accuralely recor(b his fndingson film. Addliooally, lhe pholograph shoridconv€y a stror€ estrlelic impressim to lhevis/ver Valuable records shdid mt bedocumeited with runof-thefl ill dtolo6.

To furlher inpro/e your rcsults in photcmicrography. lt h important lhal you aler4/scheck yo|lr o n pholo6. By relerdng to thefollo/ving checl@inF, liou c€n pinpoinl anyproblenE conceming liour photo6. Tll€n prcc€ed lo the second half of this sectionlilled Photomicrcgraphy Techniques.

Checking of the finished photomicrographs1. Ouosdona r€lallng to all phoiography(l)ls lhe image in focw f/eter to pages b-27)

and lhe e&o6ure prcperly adjusted?(refet to pages 4246)

(aHave din or dust specks on the sp€cimsnfound their way onlo the photograph?telet to pages 2&33)

(3)ls lhe specimen properly stained?(4)ls there uneven illuminalion? ffetef fo

pages 225)Z Cofor phologr.phy frefer to pages 47.54)(1)ls the baclloround (empty space) white or

ol light grcy lone?(4ls lhe color of he specimen accuralely

reprcduced?

nrm Kodachrome 25

3. Bfackrndwhlb photogt $if teler topages 5557)

(])Do€s the tinished photograph sfrori,graded black ard while tones wilhoulsively dark (solid black) shadow areaswashed{ur (corndelely $/hite) highlights?

1984 Dec.l

Purpos€

1

a3

a678I1 0t l121 3t 4

01 7

18

rs

a_n .aa

1 3t 4

11lq1 718

19ry21

i4an4aI31

3

. v

5

Photomicrography techniques

Exposure adjustment techniquesWhen tak ng pholom crcgraphs, exposurecompensalon is necessary depending on ihedislibUlon of the specimen in lhe iield.But ii the specimen is evenly distr buted

1. PM.lOAD(60% Average meierlng wlth mod€lPM.lOAD)

within the nlegrated or spoi meieing rcnge,no compensalion is requ red (in case oi 1X).

$ $

#

Z P-ata l

Z PM.IOADSrMod€l PM.1oADS can b€ u3€d tor inleg-6ted melering ol 3ool" anct spol melenngot 1oh.\

Measuring alaaSoecimen condlllon wlthin

the l% meledng ar€a

Brightf ield background dottedwilh fairly dense sp€cimens 0.25X

Sel lhe reclpro

lilmbelngused.

Brighlf ield background con'laining scattered specrmens 0.5x

Specimen is evenLy d slribLnedwlhin lhe I % metering area. 1X

*t

Aboul half (50%) 01 thespecimen is distribuled wiihlnthe da background

Aboui onefourlh (250lo)of lhespecimen ls distributed n thedark background

The dark backgrcund is dotled the lSo/ASAsensitivily dial

Use of the AE lockBy prcssing this button during the automaticexposure rnode, you can adjust lhe exposurelime indicaled on the display panet (expectedexposure lime, actual exposure time).

AE lock operetion

Plr.loAD (averlge mei.riig d}%)P -10lDS fnr6grat€d metoring g,%,spot mstering l%)

Wlen pressing _lhe AE 'ock hJtton, the

wuning light abo/e it nashe6 to indicateAE lock mo(b. The unil is oow ready forptDlog€phirE. To cancel this tuncttonpress the AE lock button again. Thissl|itches lhe light oft and the de\rice reverls to nonnal autornatic oocure anode.

Aller poGilior ng lhe specimen lo bephotogmphed and pressing the AE lockbutton, lhe $/amiru light abo/e it tlashesto indicate AE lock mode. Then, afterlaling the frst photograph. eecr.irelirne is locked.in.

To cancel thjs tunclion pr€6s lheAE bckbutlon again. This switches the waminglight off, and lhe device reverts lo thenorml automallc e4osure mo(b.

gimlnaling density yadatlooe by AE lock1. Taking panoramlc pictues uslng ih€

AE lockFor panorama pictures wheie any number oicopies is laken fiom differenl seclions oi thesame specimen or where several pholos arepatched logeiher, pertect panoramic photcgraphs wilh uniform density can be laken bylocking-in a lixed exposure for all the pholo

PanoEma photography

z Erample ol uslng lhe AE lock on modelPM.'IOADS

4)ln conbination with spot neteing' specirnen areas requiring spoi meteringare nol cenlered because of faminq prob€ms, lhey are firsl posilioned in the cenlerand the AE lock is activated. Then thesoecimens are moved back lo lheir orlginalaosilion and pholographed.

1 Th€ sperm€n detail ls al th6 locallonwhere it is to be pholographed, bulexposure cannol be m€suGd

2 Aller movhq th€ spocinen delailtothe measuring area and aclival,ngthe aE lock,lhe sp€clm€n d€tailcannow be moved back lo iis o.loinalpostllon and rh6 pholooraph s lak€n

2)!!9!9gEP!!!9 vt!!i! lJto naximum exposurc time

' exposure display ls c ose lo the maxrmumeroosure lime, lor example during fluores_:ence photomicrography, the pholographiceouipment mighl display an underexposure^?rning during automalic exposure. This can.€ prevented by using the AE lock, resulting. ong rme exposure. Th€ sAFEry lamp comer ro r€d .nd

an audlble warning sounds.

Compensating for a film'sreciprocity failure

W:th normally used phol€raphic emutsionsthere is a rule (the reciprocily law) thaldetermines the luminance of he light strikingthe lilm surface- According to this rute, lhetolal amount of e&osure is delined as theproducl of the lumimnce and lhe exposuretime. For example, the anoont of oocurewita 1,60 sec. o@surc at E is the iameas lor I B0 sec, at f1 1 . &Jt for longer expoGurelimes this rule no longer applies, leading touncler exposure and changes in color reproducljon, This phenomenon is knq n asreciprocily law lailure_ Bul sjnce, in d|otomrcrography, expoGure compensation cannotbe carried out via the aperlufe diaphraqm,exposure time is lenglhened or shonenedlo obtain a suitable expmure level. lt thereciprocity dial is set on rnodets PM-1OADSand PMIoAD, compensaiion is carriedoul automatically, resulling in pfoper

Ex8mple ol comp€rcallon (u,t|€n Ustng Fulhhrome ROl00)

No corn.p€nselion

Compeftsallon

Exrnd.: Dai. ior rhe afirracl€r&llcs ol r€clprodty law teilu|e {Kodrk cotor tflm DKD-|/|1)

compensation by lilter are necessary. Foradditioml dah on recipnccity law tailurecharacterislics conlact the lilm manufacturcr

exposure. lJneven color reprcdrction muatbe compensat€d for with a CC flter.'ll|e abore chan [sts lhe compensation datalor rcciprocity law failure characterislics whenusing Kodachrome 25 ilm lor geneEl ptrolomicrography. The chan shows lhat for tongexpo6ure times exceeding 1 sec., bottlexDo6ure time @rnoensation and color

Color photomicrography

Color fi lm'i.v different types oi coor i m are oiiered

:.e market today eavnu tfre Lrser 2t a:: rs 1') lrhich brand to use tor photomrcro:rhy Norma ly. day ghl typ'- an ISO/ASA speed of 50.100 s lsed

e microscopes req! re a lghl baancing' i rLBD-2. 2N)

EKachrorne 64EKachrame 100

Agfachror.e CT 18Agiachrome 50 Type SAgiachrome 100Fulchrome 50DFujichronre r00D

l. vew or these lactors the iolowng conditons can be sel ior pholomcrolraohy

! l!!qq1!(2)Good color contrasl

l?lqlgq J9!9r !9!!Iro!For noma photom crography n lrans.

m l redlghr brghl f ied uminal ion color i mwth an lSo/ASA soeed belween 50 and 100wiass!re salisiaclorv qla i1y. For specal casessJch as prnted p i rb caton or enlargemenis<cd:chro'.e fim oijers lood qla lyr.,/.e. oholograprr ng dark specmens (phase

conlrasl poarz€d ght. 1l!orescence)exmsure lime ncreases AtiroLrgh tirere.nay be ercessve gra n for cases reqLr r n-oiasl shuller soeeds lse flm willr a h ghrSO/ASA speed

Day ight typ€

t4g

::quiremenls forthe selection of colorlilmI m wth hqh resolvfg power. s fce pholo_ .rograplry requ res disoiay oi dela ed

-_! d ispay must be abe to dscrm nate

:.lween the f ne coor dilferentalons con.: . ied in a sn! e soecmen.: : lh lLr reproducton o i specmen coors

inoul backgrolnd d sco oraiion

Eldachrome 50

Agfachrome 50 lype.L

I

Types of filtersFill€rsSe ection oi filers lor use in phoiomicrcgraphy is based on lhe tlpe ot I m used.Use LBD-2N lor daylight lilms and LBT I lterslor tungsten type lilm For changing lighlinlensity, use of a neutral densiiy iiler (ND)is recommended. The number on the lllterrirn gives lhe transmssion valle In caseoi iiller ND6, 94% of rhe rotal iluminalinglighi is absofbed and ony 6% of lhe ighlis lrafsmiired.

Types of lilter and their lunctionsF ters ior lse n coor pholomicrographycomprise ihe io lowing main types:

I Lighl balancing lilter

To converl lhe color lemperalure oi ihe mlcroscope lghl sorirce to the lih being used.

eND (neuiral density) lilter

.F To feduce the lighl inlens ly without atiecllng colorlempelalure, when llluminalion is too b.ighl.

3 CC (color compensaling) liltor

For color rendlon problems such as slghl dilerences in co or hue or lading in developed pholos.

a4- Didymium lilter (marked FFon Olynpus mod6ls) To eriphas ze color, i.e. wiren trying 1o compensate

tor the insullicient inlinsic red color rendition oia llm or lo enhance the red color ofthe specimenor lhe coior contrasl ol Polaroid coior fim.

€ Heat absobing lilter To absorb heal rays emanating trom lhe jghlsource of lhe m croscope lo prevent damage toor deslruction oi live specimers. Since lhis filerlransmils a smal qlanlty of blue lighi, I maybecome necessary lo use a color.compensal nglilter (CCl 0Y or CCl 0M) \rhen iaklng co or photos.

a

Differences in color rendition dependingon the combination of fi lm and light source

sm .r _v rh.n !s n,o the llngslen lype I mol tcoors ab!s i r . l 'o toq at t s obta l^eoThe ta! I r Lrclr c:ses s ta Lrr. 10 lse:he pr..e- I r. .r r:lcf .he ,ori sc! cF

lo or rendilion depending on the combination ol lilm and lighl source

Daylishl.lype (sunlighl)

-.F are tro types 0i coor i n's d.y 9r1- 1br anced tor s ln g r i l :nd iLrr ls le f

: .1ba ancec Io 'ar t ic la gh1) .he seecl on. . idng upon ihc L l ln l so] 'ce Vr l la y.sers have erperencec ihe a sacponlmcnire r ohotos appear. ! ? lcessley red

' ! . lhey lsed day Sl r_ _ m lnder n.andes

Tungsten.type (adificial lighl)

I

Differences in color reproduction dependingon differences in color temperature

Dillerenc€s in color reproduclion d€pendingon dillerences in color lempefature

E'

>

Color l€mp€ralureCoor temperature designates the propenlesof the ight source. A blackbody radator.when healed. emanales ighl of dfferenl coordependrng on the temperature The properliesoflhe ight source can be nd caled by reieringto lhe lemperalLrre oi the backbody at a fxedter.perature evel. Ttiis ieature s ca ed coortenrperatLrre the numerica Lrnl being expressed ether rn abso ute lemperalure orin degrees Kevin (K)

Coor i lm for use n pholomcroqraphy snormaiy daylghl lype bul since the ightsource oi lhe mcroscope is a tungslen lypels coo. temperature s low (2800.3400rcand as such il is unsuilable ior daylighl fllrilwlh ts color lemperalure oi 550C6000KIn order to achleve the proper coor rendiliona ight baanc ng i ler is used, and llrelungslen ghl s convened 10 dayight

l j , ' , .

lli color temperalur". s low lhe photo shows a red sh 11 and .doi renoeralure s h qh l exhbils a'blue s '

Differences in color rendition dependingon the type of film

Color r€r'dilion for diflerent types ot film f.omlhs sam€ manufacfurcr lends to vary, ardeven ditferent orcduclion lots of lhe sametype shov/ sljght dtferences.

When you slan to lake color photo6, lirDdrould flrst d6l6mlne lhe oFffmum condilibnsto the nlm through t€st photogkphs, matchlngflem wih boh micrGcooe and soecimen(Fot the methocl ol taking t$t photosrctet to page 54)Offl€r€nce3 In color davalopftGit&pddlr! dr lh. m. ol lllmI}re 6lx ptroloG on the rlght were tak€n wllhflms ol dlfler€it brancls, and wflfi thE €xc€ptbn of lhe ISOASA speed all drotographicconditions were identical. lhis €vample dearly*lo € that properties such as color rerditior!contrasl, cladty of baclgrerd, etc. are alldlf-ftrcnt because of lhe various brards of tilm.

Photognphlc condldortBHS, PM.ioAD, LBD,{ (color tompontu|l 5500f0, LBT (color tampor.ture 3,mr+duii.. 3pe€d 0.1+l s€cD.tlglrt tyF

llrngd€.r lpe

Purchase of color film1. What lo walch lorwh€n buylng color lilmI Coor lilm is a hghy sensitive subslance,

and envircnmenta laclors such as healand humdity easily cause changes n filmspeed and coor rendition. When buyngcoor film, avoid camera siores in whichthe she ves slor ng color t m are exposedto sunlghl.

I Choose only cobr r m wiih a sufficienlylong peiod before the explraiion dareas marked on lhe package.

I Co or fendition oi lhe same lilm tlpe maydiiier, ii il cornprises dltierenl prodLrctionlots. li you lrequently lse coor film, youcan use il under ideniical photographicconditions and avo d varaiions in the coorol the t lrn if you buy arge quanlilies iromthe same production lol.

sg:im of Yalldlry (ddEnon dal€)

2. How to slore lilmFor the basic pinciples ol composl on andcolor rendition check the lechnicai literalureon lhe subjecl. As has been poinled out,perforrnance changes according 1o the con.dilions under which the f m is slored,

Color I m is normallykept n the relrigeratorlo protect it irorn the etlecls ol heat andhumidjty. Removelhe lilm trom lhe reir geralort hour before lse and alor t 10 reach roomlempe€lure. Do nol remove ihe fim lrcmlhe rc immediatey after takinq it out ol the

/-F\ /-f--\[ * , ( , r@M

reirigeralor lo prevenl condensalion on lhe

3. Poinls calllng tor sp€clal att€ntlonwhen using lhe lllm

I Do not leave ihe Jim in ihe camera longer

I Even though olher pholograph c condilionsmay be idenlical, color rendilion may varywlh low or high ambient temperalures.

I Do nol use the fim in a gaseous en-vironment (such as lorma n), since colorrcndilion wil be aclversely aflecled.

,

I

IE E :; : !

;:Eii160 2/54

I Develop exposed f m as soon as possible.

z%o

Use of color-compensating (CC) filterswhen taking color photos

S:nce colorcompensaiing lilters are highlysensilive lo heai, do nol use them close

ICC filter is the abbrevialion for color-compensaling filer, and lhe color oi iheparllcular Jilter is marked in capita letterson its rim. Six dillerenl colors are available,each color being offered in six dilferenldegrees oi densily. No. 5 and No. 10 aremost widely used in pholomicrography.

lcolor flters are made by severa nanu-'lacturers. Check your Olwpus dealer ora repuiable camera slore ior avaiiability.

:c lhe lighl source.

Color to b€ r€duc€d Color€mp€n6ating lllt.r rcqulr€d

BLUE ccYCYAN Red ccBGNEEN Magenla ccMYELLOW Blue ccBRED CYan

MAGENTA Green CCG

H

Measuing wilh CTR

0.8x, 1.25XColor compensatlon

4. Organizing photographing data[.4ake I a habil 1o record al dala rearingio bollr lesl phoiography and reguarphotcgraphy. lf the pholographic condilions lhatresulled in good pholos are rela ned on i e.lrolbeshool ng and correclon of probemscan be caried oul quickly and efiiclenly.

5. Slodng developd filmWhen storng I m ior a ong per od oi tme.the most imporlant precaulon s proteclionfrorn light Siice mold is prevalent in hot anchumd paces, t is advlsable lo keep rhe iimliglrty seaed, together wilh a desccant

We recamnend storing film in an envnon.nentwith a relative hunidiry of 1U0% andan ambient tenperature below 21 oC (71' f ).(fron Kodak Colot OKP 141 instructions).

Test photography1. The need lor lest pholographyEven ii you |]se aulomatc pholographic equ p'menl and set both lhe ISO/ASA speed andlhe rec proclly aw iaiure characteislcs,you cannol be sure that yo! wil alwaysach eve the perfecl exposLrre ior every typeol specimen The reason ior llrls s thatphoiograplrlc equpmenl s mafufactu.ed tomalch dstfbLrton state, coor and exposureslandard oi average speclmens. ln order toobraln belter resulls, you shold I rst laketesl pholographs lo match lhe conditions oithe sp€cimen, fim, nricroscope. iilers etc

3. Evaluatlng lhe test photosobserval on oi reversal i m transparenc escan be caried oui eillrer wilh a lighl boxor a slide projeclor, bLrt ihe dispiay of cotorvares wde y wilh lhe color temperalure otlhese lighi sources As a slandard ior eva uafing the pfopef coor Lrse a ghl box wilha color lemperalure close to 5000K.'Tl]e

io lowing lp€s ol lghi boxes areavaiable on lhe marketC Fuji film color box 5000

O Dlrotesl Color Classe.50O General Eecrric Chroma 50

2. Method lor test photography(when using nadels BHS and PM-12ADand reversal filn)

(1)Prepare one rol oi illm (36 exposlres)anda irequenty used spec men

(Z)Set lhe lest condilions (see chan 1) andprepare a data chait (see charl 2). List nga the condtons gven n charl 1 wil resLrllin a dala chan 2. Take yoLr photos basedon this chair and delermne lhe optmumpholograph c conditions on the basjs ol lheresulis oblaned on lhe developed I m.

No CTR Vo lase 10V, 9V 8V

Co orcompensal ng I lter Cc5G no corapensat on, c,cs\,tr

Charl 2: Data charlColor

lemperelure Exposure Color com.

12

it1 bgooK5 o ra 10v789

J9 ssoox

1' sv

0.8xNo com' -

1Xpensar on1.25X

CC5G

0.8x_1X

1.25X

cc5M0.8xl x

1,?sI0.8x'lx

'| 25X

170 mired, 180 m red, lmmred,5900K 5500K 5260K

Charl l: Test conditions

Black.and-white photography

Black-and-white film: nce the condtons diier ior plrotom crcr: aphy and generaL phoiography. i1 s:cessary to seecl lhe proper iim Jor each

'.oe oi pholography In photom c rogfaphy.!h conl ras l j m wth rne gra n is used in

-.der to document m nule slrirclures ol: oo! ca spec mens and to achieve sharp:_ololraphic reproduclon. Examples oi lhs

n type are Kodak Panatom c X. Agiapan 25.rd Pan F. and Fll Neopan F

€

,:jl r#

I I

Jse ol black-and.while liln':re are many lypes of black and.wh le ilmj the key toobla ning good pholographs esreectng the r ight f lm lor a par lcu lar job

SO/ASASpecinren

Norma or hgh conkaslior tlre slaned secllon oigenera pathoogical

palno 09 ca speclmens.as we as those ol otherareas. Wlren shape smore mpo(anl llran

Dark spec mens. andwhen ong srrltter speed

Agfapan 25on the coor 01 lhe

132502532

Kodak Technical Pan2415Agfaonho 25

Srce the conlrast oi the filmis very lrigh, the range or Proper exposure is very narrcwUriless expos!re is sel wlhln+l/3 slep oi lhe opi murnlhe pholo w I be ether over

64

1002532

Agiapan 400l lord HP5

X33 u"" t ese ryp"" *r,"n vo,400 want 10 shol(en exposlre tme

5G1600 a1lhe expense of f lm g lan400

tComparison of different film brandsTypes oi fim orovding a wde gradalon iomshadow ro h 9lr _ohts and featurng goodconlrast are Fll Neopan F and KodakPanalomc X. Technica Pan oiiers s ghtyh gher contrasl wh le N4 n Ci]py erim nales

Neopan SS and Tr X on lrre olher hand.have fanly o!\r conliast

Kodak Techn car Pan 24t5

FiltersUs€ ot contrast lillorsConirast fllers are used io conlrol lhe conlrastor b ack.and'white pholos.For black and while pholomicrography, greeni ters are normally used, bul choce oi lhemosl effective flter depends on lhe tlpe ofspecimen. Filterc lhat enhafce the colorsand conlrast of the specimen are:

Color ol lh6 Color ol rh€ tllt r

GreenYellow/orange Blue

Blue Oranqe

Ellects ot verlous conlrasl lillers

the red po.lion is €nhanced

For reducing conlrasi, use a Uller oi lhe sarne.olor as lhe spec men.

Reasons for using a gr€€n lill,ar-iere are two reasons why conlrast improvesshef a green llter is used.T)S nce objective abe(ations are rnosl

effectively compensaled near lhe greenwavelenglh, loss oi image c arity due lochromatic abefialion is avened by a greenI ller.

2)Dyes such as hematoxyln and eosinabsorb green lighl well, resulting in higherconlrast when a green llller is used.

lhs r€d Porlion is reduced

Photography with Polaroid@ filmOne of the acl€ntages of Polaro d litm is thalI can be quick]y vlewed as a iinished pholo,bul il is seldom possible 10 oblain cotorreproducllon comparable to thal oi 35mm

by'haPora|o]dcor@ralioi 'camb.ido€.MlEs'U's.A'

2. Photogrrphlc t€chnlqu€sl1J B I ac kand -wh i t e p h o t o g h p h yUse a green filter to obtain good contrast.(2)Colot photogftphySince lhe overall color hue tends to be eitherighl green or blue, use CC10-20M or

CC10-20Y lor compensation. ll you wanlio enhance color conlrast, use the FF iilleravailable trom you. Olympus dealer.

Since Polaroid iilm is morc easity atfectedby reciprocily law Jailure than normal iilm,exposurc lime should be adjusted 10 belween0-05 and 0.5 seconds.

ol lllm us€d ln r.Pnyt 1E' xS th'

(actuel slzo 73crn x 9.5cn) ,l'x5" (actual da gdnx11.5cm)

I

kIanI

I

Type tso/AsAspe€o

rack containin!8 photos Tlpe ISO/ASA

speed Sheelrack coolaining

I photos107 3m0 o 52 4(J0 o67 3000 o 30m o

665(PdnU

negalave)

50 o4m o

55(P nU

negative)

50o

I108 80 o 58 80 o668 80 o 59 80 o669 80 o 559 80 o

14'x5' film hoLclers diller according to lhe type ot titm

sh@i lllm-:(s type i m hotd€r Packed fr/m-550 typ€ tllm hotdef coror lemoeraiure 4500K-LBE!2N olus FF tllto.

SectionSHow to obtaln good prinis

How to ob ta in oood co lo r p r i n t s . . . . . . . . . . . . . . @-61aPreparing blacLand-white brlnts . . . . . . . . . . . . .. . . 62

Maiks on the Dhoto . . . . . . . . . . . . . . . , . . . . . . . . . . . . . . .63How to avoid marks during development .. . . . . . . .64-65

How to obtain good color printsThe lollowlng diagEm shows how to getgood color pdnts {positiye pdnb).

Color negalive lilm

ln pholomlcrography. lhe color reprodlctionoi lin shed pr nts taken wllh negal ve fi mdiffers irom the observed color-a lact manyusers are probab y very much awa.e of.Possible reasons for this are thal the technicianwho made lhe pinis d d not view the specimen color lhrough a microscope and thusdid nol know lhe actual color, oa the pholcgapher made a mistake when he lsed a f llerand lailed to notice ihal lhe iilm qualitydeteriorated. In ordef lo gel beter resulls,you should lirst photograph lhe specime.trom which you wanl lo get prinls on reversallm, and atlach lhe iinished photo as cojorreterence sample 1o the negative lilm. Pinlscan also be made lrom color rcvercal {ilm,bul lhe process does not match the pintquality as obtained from negative lilm.

R6wtsal fllm

a

Color pdnl with Poor color r€produclion

Color pdnt with good color reprcduclion

Preparing black-and-white printsThere are many technicat pubtications on ihemarkel llrat expta n in delait how to devetopiilm. li you plan to do your owr iitm processrng, use lhese books as reference.But since conlrast in photomlcrog.aphy isower than with normal phoiographlc subjecls,you shorild pay panicular attention lo under.developmenl. ll you make a positive printirom an underdeveloped, /ow-density negalive,an even grealer reduction in contrasl witloccur, badng lo a poor resuti.

1. How to obtain a good negative(])ljse a devetoper thal malches lhe iitm(2)Use the developer allhe lemperalure

specilied by the manuiacturer(3)Pdhere lo the speciiied deve opment iime

lhal malches llm sensil vity(4)Agilate genlly and freqlenlly to eliminaie

uneven oeve opmenii5)Make sure lhal lhere is nelther loo much

nor loo I tl e it ng(6)Follow the paper specificaliols when

washing the prinlOHandle ihe negative carelutty and pfolect

1 lrom fingerp|nts, scraiches and dusl

2. Selection of phorogEphlc pap€rWhen making pints, you can vary tlte contrasldepend ng on lhe t 9€ of pinting paper.select the lpe that best suits bolh thedensity of the negative and sp€ctmen contrast.The examples on ihe fight show prinls madeirom lhe same negative when using difierenttlpes 01 pinling paper.

Grade no olphotographlc pap€r

Grade 3 nomalcontrast

t€

Marks on the photoMarks on the print tend to sland oul moreprominenlly in pholomicrography than onnomal pholos ol p€ople and landscapes.Since pholomicrcgraphy empoys lilm withfaiiy high conlrasl, even the rosl minute differences in brighlness show up in the piclure.Marks resu I ng from improper developmentare also very conspicuous. This seclion dealswth marks resulting lrom developrnenl.

1. Ma*s as a esult ol negaliv€ d€veloFnent

Both uppd and lower ponbns of lho photo arebnohr* than lhe central section.

How to avoid creating ma*s(1)Confirm the degree ol exhaustion ol lhe

Carelully note producl on dale and lre.quency ol use of the deveLoper

(2)Agilale the developer lhorough y beiore Lrse.(3)Use the developer al lhe specified

(4)Do not work wlh an exposire lhat reslltsin a developmenl lime of Less lhan 5 mlnules(lhe resull wil be exlremely clesensitizeddevelopmenl).

(s)Agilale lhe baih thoroughly during deve oF

(6)Mainiain lhe speciiied lixng lme.f)Wash lhe film thorough y.

Marks with iregular b qhlness.how up.

It you use an aLnomatic processor to developlhe prinling paper no marks will occlr, butslnce mar!€ can easiry sho / up whendeve oping in a t€y, rhe io lowing poinls

How to avoid crcating na*s(1)Agilale lhe develop€r lhoroughly.(2)Keep the lemperalure of lhe developer at

20'C. or as speciiied by the manuiaclurer.(3)Since marks can easily occur if deve or

ment time s too short, set the exposureof lhe en arger al such a evel that develoPment tirne s between I min 30 sec and2 min.

(4)Agitate the balh thoroughly during develotr

(s)Conl nue to agilale lhorclghy even whilestopping and iixing.

(6)Wash the pinl lhorcughly in runnlng waler.Regularbase paper 30€0 minResin-coaled paper- 4.5 min

2 Ma*s occuriog dudng prinl devglopment

How to avoid marks during developmentDoYslopment ol tllm

Snce ma.ks on lhe lilm occur easily whenuslng a ree -tlpe lank, developmeni shouldbe done in lhe io lowing maaner n the

Development ot pdntlng paper

This secl on deals with developmenl usinga tlay.

t,21

lmmerse lhe ree in lhe developer, and aiterturning ii 2'3 limes, lap il anolher 2 3 limesaga nsl lhe botlom ol the lank to removea r bubbles sllckng lo the fim surface.Complete this process wilh n 56 seconds.

.lI

Whie holdlng lhe edge ol the exposed pr nlingpaper lighlly wilh a pair of longs, quicklyplace il inio the developer.

Now remove the reel lrom lhe developer andmmedialey immerse il again. Afler rcpeal nglhs procedure, iurn lhe ree immersed inlhe developer 23 times, then pause for30 seconds.

lAs shoM in lhe piclure, do noi immerselhe prinlng paper inlo the developerparallel to lhe developer surface, bU tillil nlo the deve oper balh.

2

'@@@

X

/ - - - - - - - \ -

Alter completing developmenl, lollow lhesequence of stopping, iixlng, and wash ng

Thodeveloperwill nol b€sutficiently aoilared simply bytuning lhet@|, and the resullwill be spors on ihe lilm,

Pedorm lhe operal on shown inol developmenl work.

I Make sure thal lhe photograplric paperdoes nol float !p lo the surface of lhe

After complelifg development work on lhepr nting paper, qu cky pe.lorm the sequenceol stopping, fixing, and washng.I Make sure lhat separale sheels ot prifilng

paper do nol stick together if the fxerI Do nol use the same pa r of iongs during

slopping and lixing lhal you rsed du.ngdevelopmenl work.Make sure nol to codaminale the develoPef wth e lher slop baih or I xer qu d.

ro agilale the phologGphic paper, ho d ilat a corner wlh a pair oi longs and mover back and lorlh whlle I is submerged n lheaeveloper. Repeat this operalion in 15-secondnterva s unilldeve opment work is comp e1ed.

By @movlng lh€ rcelfrom lhe dev6lop.r,lhed€v€lops can reach lh€ spac€s between theroll6dlilm, p€vontlno lhe oon€nling ofspols.

piclure 2 of ihe left in 3o.second nlervals I I the compielon

Section4Trouble-shooling

P|obloms in linished pholos and lheir con€ctionPoor color reproduction . . . . . . . . . . . . . . . . . . . . . . . .68-7'1B l u r r e d i m a g e . . , , . . . . . . . . 7 2 - 7 3The image is in focus but not sharp. . . . . . . . . . . . . . 74-79Objects other than the specimen image appeared

on the t i lm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .80-81Uneven b r i gh tness . . . . . . . . 82 -83

Prcblems in finished photos and theil conection

-Poor color reproduction1. Th€ beckground is coloEd (.€dibhF)

Caul6 Cor'rctlon Rama*s Rel€r loDaq€

The color temDeralu@ of the lliumi-natlon ls not malched to that ofthe fikn..Lioht balanclng fllter specified by

manulacturer ls not us€d.

.ljse flltels sp€clfled by the manu'lacturer.

.Daylisht type: LBI!2N, 5,5mK,D settlng

.Tungst€n type: LBT, 34mKT setting

Page 48/page 50

Lamp \oltage ls too low (loo high).

6v-rcltege ls t@ low {Bd hu6).

l1v-rcllaoo l. too hloh {bluo hu€).

.Raise (low60 lamp voltag€ .BHS above &5V.BHT and BHTU at 5-6V.lf you have reached the desired

color temp€ature, do not chanoethe voltage position. lf you wantto chango lrieBity, u6€ an NDfllt6r.

.!!b recommend that for iestphotograplry the voltage shouldbe set lo the standad Dosition+1V whgn fi6t detednininglhE ootimum color temDeEturc.

Pag6 5(ypage 54

Blackenlng of lamp due to prelonged Uae

.Replace wlth a now lamp

@

2. The background is color€d (gr€€dmagenta)

Caus6 Con€ction R€marks R€f€r toDeq€

.A iilm dilferent from your usualfilm lype was used.

.Since color rendition varies withdifferent film types even from thesame maker, choose a lilm withcoloring suitable for your type

Page 51

.A iilm of the same type but withdifterent emulsion number wasused,

.li possible buy film with thesame emulsion numb€r in laryoquantities.

.ln order to maintain the perfor-mance quality of the film, sloreit in a relfigoralor and removeit one ho0r prior to us6, allowingit to reach room lemperature,

.Even for film of the same typethere will be slight variations incolof reproduction depending onlabofatory, development condllions,and staining of the specimen.

Pag6 52

aa

Ota

at r t Io C

t t l trt

.l.Jse a color€ompensating (CC)lilter

at

O-

at a r IeB

a +t t l t

racc05M-optlmum compensallon

aa

Ort : i .

r . l r

aa

*

.acc10M-ov6r{ompensared

'Color€ompensatin0 (CC) tlltersBuy from your pholo dealer,

.Too much gr€€nUse CcM {mag€nta)

.Too much pinkUse CCG (gr€en)

For photomicrography, use ccfilters 0+10. For lurlher compensa-tion use CC05 and CClologether

Page 53

3. Inconecl color rendilion

Caus€ Con€ctlon Remarks Reler lopag€

.An excessively long exposuretime has been used and the char-acieristics of reciprocity lawfailure lead io incoftect colorrendition.

.Set shutter spe€d at 0.01{.05sec.Adjust shutterspeed uniJormly asfal as possible by using an ND{ilter.

.When using long exposure limes(above 0.5 sec) wilh modelsPIV'1oAD and PM'1oADS, set thecharacterislics number of the filmused on ihe dial for reciprocitylaw failure,

.With long exposure times, evenwiih the exposurc time compen-sated, the color fendition changesas a rcsull of film properties.Use a colorcompensaling filterspeciiied by the film manu'

Page 216

.Automatic exposure has beenused withoul adiustment.

.When using models P[,]-1oADS/PMJoAD set the exposure adiust-ment dial lo:0.8-0.25X for bighl backgroundspecimens and to 1.2t4Xfordarkbackground specimens.

.Carry out exposu€ adjustmentsBrlght backgrou nd: ISO/ASA 100 to 50Dark background: ISO/ASA 100 to 200

.ll you change the magnificationof the objeclive, the light distribu-lion within the visual field willatso cnange.

Page 4Zpage 43

.Aclual and nominal sensitivityol lhe iilrn differ

.Vary the ISO/ASA speed setling. .Nominal speed may diiJer lromactual light sensitivity as muchas 1/3 to 1/2 aperture.

Page 42

4. Poor color ol a prinl enlarg€d lrom a negative

.The printing technician in thelaboralory is nol sure of lhesubiect's correcl color.

.Add a lransparency of the samespecimen with correct color asa sample.

.Better coior is achieved whenshooting a transparcncy withreversal color film (but the costof pints goes up and quality

1

5. Poor color ol Polaroid lilm

Cau3e Coneclion Remarks R€fer to

color temperature. .The film is a daylight type, butbecause of the film characteris-tics color temperature shoutd beset lower than for 35mm film.

.lf you s€t color temperature b+tween 2rc00K and 5000K anduse an Olympus FF filter, colorcontrasl will b€ enhanced.

Page 58

.Abnormal color characieristics. .lf the abnormalilies are withinthe adjusiment range lor colortemperature and within theoperalional range ol the color-compensatlng lilter, determineihe conditions by test photography

.Since Polaroid film is easilyaftected by adverse storageconditions as far as color charac-leristics are concemed, protect itirom heat and humidity bystoringit in a refrigerator.

Page 54page 58

.Term of validily of the lllm has .Use film whose term of validityhas not yet expired.

.ll room temperature is abruptlylowered (raised), sensitivity isalfected and print color turnstowards blue ted).

.For developmeni time and colorreproduction, refer to the filminstructions,

Blurred imagel. The ov€rall locus ol the oicturc is blun€d

Cau3€ Conlction Remarks page

.Finder 6yepisce or focusing t6le-scope is nol prope y adjusted,

lt

ll

.Adjust the diopter until the doublecross lines are clearly visible.

.Slnce most people do not havethe same visual acuity ln botheyes, d€l€mlne which eye youalways use for focusing.

Pages2e27

.Blur and dritt resulting fromvibrations.

lm.o..fi6t.d by vlbEttons

.Uss a vlbration.proof table.

dF

.Uss an ND fllt€r and lncreaseshutter speed (1/2 sec to 1 sec).

.Use a stand for the photographlcequipment to separate photo-graphlc equipmenl and microscope.

Page 12paqe 48

2. Focuslng enor occurs wh€n you u96 a low.magnlcallon obiective ol less than 4X

3. The p€rlph€ry ls unilomly blun€d

Ceus€ Coar€cllon Remafts Reiar topa96

.lf magni{ication is low, tocaldepth at the lllm plane becomesshalloq easily causing erorsin focusing.

.Mouni a locuslng magnifier tolhe f Indereyepiece or the focusinglelescope, and after focusing onthe double cross lines, fine focusthe specimen.

.lncreasing the magniticatloneliminate6 tocusing erors,

Page 27

Cau3€ Coreclion R6ma|ks Reler topaq€

.An Achromat type obj€ctive wasus€o.

1.,iil;.' l,r,i';.,r',;';iiii,''''r,;ii'':: ' , : ,1:,11i,; i ; , , . ,

ause a Plan Achromat typeobiective

;;1..i.'i:.1 ;i:'-:: l,i;"li ;1/ : : : ) . \ . ' - - : : ' . ) : t t . : :

:: .:':i tl: i: - r 11.;'i j;:. : .ij,

.:r '::;j i liir'i'.1.t...'1,.: ':i

; t : : ' . . - . _ : J : - - i : . . ! : - : r .

. .;.,. .: .i!:::.:'1:.:):.t .

D Plan 1oX NFK 2-5X

Pag€ 14

.Objective and photo eyepiecewere usgd in th6 wrong com-bination.

.LB series (long banel)-NFKphoto eyepiece

ashort-barrel serles-FK photoeyep|ece

Page 13

The image is in focus but not sharp1. Inadequat€ resolving power

.Use ol a combination of low-magnification obiective and high-magnification pholo eyepiece,

.ln order to obtain high resotuiion,use an objective with largenumeical aperture and a pholoeyepiece with low magnification.

-? ,1

.ln order to obtain a magni{ication;i ioot i; tniirr. pti ", ,iJ- Pase 38either an objective zlox with aphoto eyepiece of 2.5X or anobjective 20X with a pholo eyepiece 5X. To increase resolvingpower, the combination of obiec-tive 40X and photo eyepiece 2.5X iis pfeferabie. (Focal depth,however, will b€come shallow)

.Use of the condenser with lheapenure iis diaphragm fullyopened.

.Stop down theaperture iris dia-phragm to 60€0%

.Vary the amount by wh;ch youreduce theaperlure iris diaphragmaccording to the magnificaiionof the objective and the contrasiol the specimen.

.The field iris diaphragm was fullyopened,

stray light, slopdown ihe field irisdiaphram to anarea only slightlylarger than lhe

.Do not reduce the diameterof lhefield kis diaphragm to such anextent that it louches the framereticle because lhe aclual areaphotographed is always slightlylarger lhan the area wilhin thelrame reticle.

.Use of thick cover glass. .Use a cover glass with a thick.ness of 0.17mm.

' : '-:-. ,,. ' ' ' .

' - : . = ' . . . ' . t

.Olympus objectives lor biologicalspecimens have been designedin such a way ihai optimum re-solving power is obtaaned whena cover glass with a lhicknessof 0.17mm is used.

Cause Cor€ction Beler lopag6

.The specimen slain is too weak,resulting in lack of conlrast.(Photographing a specimen thatcannot be stained)

.Use a denser stain,

.Use a contrast lilter (fof black-and.whiie pholography only).

.ll the specimen cannol be slained, use phase contrast, differential inteFiercnce contasl or darklield to create conirasl oDticallv.

gee

G*,iloiii€Gnlial inlorf ercnce conlrasl

ST _Effiil

Page 57

Cause ConEctlon Remarks Ref€r topeg€

Color photography

.Use of a low convasi film, .l.Js€ a high conlrast fllm.

:', r-'d):.:')*.:;i;

tt))b,;nite-:ii:);'.':ii;i;?;":i

Page 51

Black'and.whlte photography.Variations in lhe so€clral s€n-

sitivity of the film ifiect ihe.Normally, a green filter is used,

but if you want to emphasize asp€cific portion of the sp€clm€n,use another contrast flller,

.lf a filter complementing the colorof a specimen is used, it willemphasize lhe contrast.

Page 57

2. The image appears hazy

Caus€ ConEctlon R€marks Rolor topagsaThe conection collar of the ob-

iective is not adlusted to thelhickness of the cov€r glass,

, i r , - ' r t

, . ' J a .

. .

.:-1 'l: j

i. ,' '-r',!! ba' . r

. ' l r .

.Adjust the conection collar while€xamining the specimen and setit at a position providing a cl6arimage.

.On the LB objective a correctioncollar is mounted on the S PlanApo 40X S Plan 100X dry andD Plan Apo 60X.

Page 16

Cause Con€cllon Bema*s Relgl lopags

.An objective normally used wilhcover{lassed specimens wasussd on a specimen withoutcoverglass (or vice vefsa),

.Use a no{over objective.

' ji;t, ) \.'1t -.J

)/ '-. J .'1\)/- '\

. 'U . . ^ \J \ - -' , 4 ) ' ' ( t - \)- - !i !.-- - - , , r ' - - \ - '- 'J .-. t .

' ,-- '( . v:

LB objectives.NC S Plan 40X.NC D Plan FL mX.NC S Plan Apo 1mX oil.NC S Plan 100X dry

Page 17

.Finqenrints, an oil lilm or dirlparticles in the optical syslem(objective front lens, photo eye-piece, prism, specimen, etc.)

.Clean lhe oplical system.

.Always coverlhe microscope witha dust cover when not in use.

.lt you use dry 40X or 60Xobjectives together wiih an oil.immersion objective, ths oilon ihespecimen may soil the fronllenses ol lhe dry objeciives.

Pages26- 32

..,t,

. 1 :

, ' , . '

3. No sharp imag6 is obtained wilh a 10OX olflmmerslon obiective

Cau36 Conection Remarks R€l6r topage

.No immersion oil was used, .Use oil specified by the manu-facturer, Page 18

.Unsuitable oil was used, .Use specified oil, since the typesused for fluorescence and nomalwhite light often diifer

Page 18

.The oil contained air bubbtes. .Apply th€ oil aft€r you hav€removed bubbles ln the botlle.

.Remove lhe eyepiece beforeexaminalion and look through the

Page 18

.Using oil in a room with unsuitabletemperatures (too high or low)

.lf room temperaiure is either toohigh or ioo low, o. if the air is toohumid, the diffiaction indsxchanges, causing changes in theimag€, Use the oil at a roomtemperature of 22-25"C and ata humidily of about 56%.

Page 12

.The specimen is too thick. .lt is advisable to use a specimenwith a thickness of 2'3r.

- l ,

r1S:. -j- p'.o'*. !.'l

4. Entire roll ot black.and.whlle lllm ls nol sharp

5. Th6linishgd pdnl app€ars grainy

Cruse Con€ction Rema*s Reler looaQ6

.Possible causes are: type of film,emursrcn, overexposure, over-development, improper handlingor accidents during development,

.Check lhese possibililies and Pages

B{- e,5

Caus€ Coreclion R€mark5 Refer lopaq€

.A fllm wlth coarse orain was osed, .Use a fine grain film. .Kodak Panatomic X.Kodak Technical Pan 2415

.Agfapan 25

.lliord Pan F

Page 55

.A slandard develooer was used. .lJse a line grain developet. .Kodak MicrodolD-23D-25

.Fuji MicrcFine

.Magnitication ratio was too high. aL,se a largsformat film. .4" x5" sheet film.

Objects other than the specimen imageappeared on the film

I

'1. Shadow.llk€ lmag€

Caus€ Con€ction Remarl(s Feter toDaqe

.Optical palh selector ot thepholographlc attachmgnt or vinccular lube was intenupled at

.Engage the optical path selectorat its prop€r posllion,

.The lield kis diaphragm wasstopped down too much.

.Open the field kis diaphragm alittle widerthan the photographedarea ot the linder eyepiece ofthe focusing telescope,

Page 23

.Tlny bits of film, dirt, etc. stuckto the p sm of lhe photographicequipment or to the largefomat

.Ch€ck for, and remove, dirt fromthe prlsm ot the photographicatlachment while the shutler isopen (Iime setling).

.Flemove the largefomal relaylens and clean it.

.Periodic checks are recom-mended if a large number ofphotographs is taken,

Pages2931

Cau3e Corectlon R6ma*s R€ler loPage

.Dirl In the optical system alocate the dirt and remove it. .Yo0 can locate the dirt by movingand rotating each checkpojnt,alt€nately looklng through theblnocular tube, the focusingtelescope of the photographicattachmsnt and the lllm plane(by placlng a piece of frostedglass in the camsra body.)

Page IPages

30-31

Cause Con€clionR€l6r lopage

.When moving the film or whenunrolling the backing paper ofthe fllm, static electricity causesspafis.

.Do not rewind lhe film too rapidly.

.Keep humidity at zl5% minimumin the room where you handlethe tilm.

aMake sure that the camen backand the darkroom ars lree of dust,

3. Rellocllon ol window or room illuminatlon

Cause ConeclionRefer to

Pag€

.Stray light gntsrs from the ey€"pieces or lhe focuslng lelescope.

.Move lhe oplical path selectorof the tdnocular tub€ io lheCamera 1m7o posilion and coverthe locusing telescope of thephotographic atlachmenl wilha cap.

aPut caps on both eyepl€ces andthe locusing telescope of thephotographic atlachment.

.Set up the mlcroscope in adlllerent location.

Page 12

Uneven brightness1. lJn€yen areas occu. on one sld6 of th6 fram€, in lhe c€nt€r, and under the pertoratlon of th€ film

Cause Con€clion R€ma*s R€ler lopaqe

.Th€ microscope light source Isnot prope y centered.

.Prope y adjust the light source.

.The field iris dlaphragm is offaxis.

.Clos€ th6 fleld his diaphragm sothat it appears in the visual fieldand adjust the conden6er tocenter the diaphragm.

Page 21

.The optical syslem is contami-nated by dirt.

.Clean the optical system. Pages29-31

.Dgvelopment problems (on black-and.whlte pint)

.Develop the film prcperly. .8e aware lhal a change In l6m-pe€ture occurs between thecenter and the pedphery of thetank as a result of hgat conduc-tlon by th6 sl€€l iank

Pages64-65

@

2. Ma*s on the nogative

Caus€ ConEction Roma*s RaLr lopaEe

.Fixing time was too short or ex-hausted fixer was us€d.

.lncrea6e tixing time or use anew lixet

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