Howto lmprove Photography Through Microscope the

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Howto get the mostout of this booklet

Thisbooklel divdedintofourman sections. ls ltemsmarked witha a I st the minmuminformationthat newcomers the art of photomicrography to know, to need whie llemsleft unmarked are addressed users to whohavealready takenphotos the mlcroscope are notsatisfied with but withthe results. relerring sections to 3 for picture By to 1 taking and section for checking, 4 youwill findthis bfochure reliable the nexttimeyouwantto takea photomicrograph. a help
lvlicroscopes suitabletor photomicrography. . . . . . . . . . .8 . Equipment suitable photomicrography. . . . . . . . . . .9 tor . .. Various typesot photomicrographic equipment a n dp e r f o r m a n c e ...........10 aEquipment needed photomicrography . . . . . . . . . . 11 for .. . Suitable localions setting thie for up microscope .......12 aObjectivesand photoeyepieces suitablefor photomicroOraphy ..........13 aDifferences the peripheral in imagesdepending on obiective sion de .. . ... . .14 aDiffeiencesin re-solution depending lne lype on of objective . . . . . . . . . . . . . . . . 15 asome hintsaboutthe objective.. . .. . . . . . . . . . . . . 16- 18 . Selection a condenser. . . . . . . . .. .. . . . . . . . . . . . . . 19 of .. a o b s e r v a t ip rn c e d u r. e . . . . . . . . . . . . . . . . . . . . . . m - 2 1 o o .s a H a n d l io f g p e c i m e . . s . . . . . . . . . . . . .. . . . . . . . . .. 2 2 ns .n . Useof the fieldirisdiaphragm. . . . . . . . . . . . . . . . . . . . .23 . lJseof the aperture diaphragm iris and its effects . .24 - 25 aBasic focusing methods. . . . . . . . . . . . . . .. . . . . . . .6 - 27 . How to spot dirt and specksof dust in the optical systemsof the microscope the Ohotomicroand g r a p h acl t a c h m e n t i .........28 aHow to cleanthe microscope frame . . . . . . . . . . . . . . . . . 29 a H o wt o c l e a nh eo p l i c a lv s t e m . . . . . . . . . . . . . . . . . 3 0 3 l t s a H o w t o c l e a n n o i i . i m m e r s io b j e c t i v e..... . . . . . . . . . 3 2 a on aHow to cleanspecimens . . . . . . . . . . . . . . . . . . . . . . . . . .33 .

FrompEparation observation to

FrompEpaaation to obseryation

Sectionl

Whenlakingpicturs in photomicrography

Sectlon2

Howto obtain good pdnls

SectionS

In the caseof poorphotos

How to obtain good prints

How to obtaingood color prints . . . . . . . . . . . . . . . . .60- 61 p aPreparing black-and.white nts . . . . . . . . . . . . . . . . . . . . .62 N l a r kosn t h ep h o t o ...........63 H o wl o a v o i d a r k s u r i n g e v e l o p m e n.t. . . . . . . 6 4 - 6 5 m d d ..

Trcublehooting

$ection4

When taking picturesin photomicrography

tr:oJclion ol eouiDmeni suilable lor GE-Jal on andphoiomicrcgraphy, basic lti.dge, observaiion procedures, cE rc methods, in etc. are explained .esylsundeGlandmanner.

l*,
l-r"= lt<r,

t"*'**

p'ocedures neededfof pholG drfferencesfilms purchase in and

and exprafanon or

can +ct processing ruin lhe best ' This seciiondealswilh film de nl, how order lo color elc. Prlnls,

Basicinlomalion in photomicrography Operating instructions modelsP[,4-1oAD for and P[,4 IoADS(with35mmcameraback) . . . . . . . . . . . 36- 37 hiu .i M a g n i f i c a toi f p h o t o g r a pe q c i p m e.n . . . . . . . . . . . 3 8 on Differences resolution in according the combinaiions to o f o b i e c t i v e s e y e p i e . .e . . . . . . . . . . . . . . . . . . . 3 8 and c .s (effective magnification Settingot photographic m a g n i f i c a .t . . .n . . io . ) ..............39 F r a m i o fg h e p e c i m .e. n . . . . . . . . . . . . . . . . . . . . . 3 9 n t s . C h e c k i n g t h ef i n i s h ep h o t o m i c r o g r a p h s . . . . . . . . 4 0 - 4 1 of d Pholomicrography techniques . . E x p o s ua d j u s t m e e c h n i q u e.s . . . . . . . . . . . . . 4 2 - 4 3 re l nt A ..........44-45 U s e f t h e El o c k o or r y C o m p e n s a t ifn g a f i l m ' s e c i p r o c if ta i l u r e . . . . . . . . . . . 4 6 color photomicrography ...........41 O C o l of r l m i OT v D e sf i l t e r s .............48 of 6 Differences colorrendition in depending the on of . combination tilm and lightsource . . . . . . . . . . . . . . 49 E Differences colorreproduclion in depending . on differences color temperature . . . . . . . . . . . . . . . 50 in on a Diflerences colorrendition in depending lhe lype ........5'1 o ff i l m . . . . . . . . . . . . . . ........52 a P u r c h ao fe o l ofri l m . . . . . . . . . s c (cc) Useof color-compensating filterswhentaking .....-.........53 c o l op h o l o s r .............54 a T e sp h o t o g r a p h y l phologEphy Black-and.white 55 a Black'and-while . tilrn 56 aComparisonof ditterentfilm brands . . . . . . . . . . . . . . . . .56 57 4 F i | t e .r. s . . . . . . . . . . . . . . . . i ........58 P h o t o g r a pw iy hP o l a r o if dl m . . . ht

Trouble-shooting
ranging linishedphotomicrcgnphs you Cngto theircharacteristics can :- causesfor iailureand take core action. In many instances, the

shoollngseclion can also serve

Problems tinishedphotosand thok co.reclion in .. Poorcolorreproduction . . . . . . . . . . . . . . . . . . . . . .68-71 . . . . . . . . . . . . . . . . . . . . .72-73 image Blurred i b T h ei m a g es i n l o c u s u tn o ts h a r p . . . . . . . . . . . . . . 7 4 ' 7 9 imaqe appeared Oojecls olherrha" the specimen ...............80-81 i o nt h e i l m . . . . . . . . . . . . 82- 83 lJneven brightness

Sectionl

From prparalion obseryation to

lvlicroscopes suitable photomicrography .. .. .. .8 for .. ... Equipment suitable photomicrography.. .. .. .. .9 fo. ... .. Various typesof photomicrographic equipment and erformance o ...........10 aEquipment needed photomicrography . . . . . . . . . . 1'l for ... Suitable locations setting the microscope . . . . .12 for .. up and aobjectives photo eyep;eces suitable for photomlcrography ..........13 in lmages depending aDiffrences the peripheral on o b j e c t id e s i g n ve ............14 in depending the type on aDifferences resolution of obiective. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 . h a . a s o m e i n t s b o utth eo b i e c t i v e . . . . . . . . . . . . . . . 1 6 - 1 8 S e l e c t io f n c o n d e n .s.e .r . . . . . . . . . . - . . . . . . . . . . . . 1 9 oa . . . a o b s e r v a t ip r o c e d u r.e.s . . . . . . . . . . . . . . . . . . . . n - 2 1 on . .. a H a n d l io fg p e c i m e .n s . . . . . . . . . . . . .. . . . . . . . . . . 2 ns Useof the fiefdirisdiaphragm. . . . . . . . . . . . . . . . . . . . .23 . useof the aperture diaphragm its eftects..24-25 iris and focusing a Baslc methods . . . . . . . . . . . . . . .... .. ..26-27 .. Howto spotdirt andspecks dustin the optical of systems the rnicroscope the photomicrc of and g r a p h i ct t a c h m e n t a ,,,......m the frame. . . . . . . . . . . . . . . . .29 aHow to clean mlcroscope the syslem . . . . . . . . . . . . . . . . 30- 31 . aHow to clean optical to clean oifimmersion an objective. . . . . . . . . . . . .32 . aHow specimens . . . . . . . . . . . . . . . . . . . . . . . . .33 .. aHow to clean

Microscopes suitable photomicrography for

acceptsthe pholomicrc 1. Thephotolub graphiccameraattachmenl.

2.

-Ihe

image can be focused throughthe binocul lube.

matching lighl 3. Thedesired inlensily tre specimen conditions be set cn by the odical palh seleclor,

4. Objectives shouldhavehigh resolution (iat andgoodllatness imageall the way to lhe periphery lhe Msual ol fielo.

5. Theslageis rolalable. is wilh 6. Thecondenser equipped an aperlurei s diaphragm cemeiing and devicelor aligning the oplicala.xis. 10

z$""[if;ffffi;'lXll""*'ipd-r'h
lighl intensity assured. 8. Sufiicient is Lightinlensily adjuslable matchthe is 10 observalion pholographing and condilions. lJnilormillumination from low 10highmagnifiction. 9. The microscope slandis resislant
again$ enemal vibralions.

------

'The phoio shoE m.del BHS

10.Provision insenion filters lor of

Equipment for suitable photomicrography

accepls35rnm 1. Thecrnea attachmenl or large{omatfilms.The35mmcamsra fllm with automatic back is equipped

lo 2. Theoplicalpalhcan be changed condilio.rs. matchphotographic arca cn be changed 3. The measuring (lntegraled to matchthe specimen, 1%) metefing 3oor-sfDt meiering is 4. The cameraattachment fmly clamped the microscope, on colortemperalure. 5. Device measuring for 6. Widerange setting for lso/AsAsensilivity

for 7. Coanpensation reciprociylaw failure for is carriedout automaljcally long adjuslment, matching 8. E\poure is spcimen conditions, possible. is 9. Manual exposure possible. applicalions. suchas 16mm 10.Special pholography, cine and35mmlime lap6e can be performed. (A specialcorfrcl unil is reqrifedin
I''rne phoroshm mod PM.1oAos.

lock mechanism lock) 11. An exposure {AE is built'in.

r ii
Various types photomicrographic of equipment performance and
Types pholomicrographic ol oquipment

E+

f -'

,'i

Pi|nOADS/

E se ol locuslng Photographic quallty

' Concernlng ioclsingscreen the ior olympus system oM cameras: TTre screen bechanged sul dflerenl can to (OM.]OM2. Olv-3. 4) Forpholo uses OM The',ocus screen ng mLrsl rebe lse No placed pholomicrography' ' mcrography screen 1 12 ior Sh|]lter !r mayoccufli magnificaton b s increased When usng40Xand100X obleclives. adlust lightinlensty llrat lhe so Shutler urmayresull b when 40X lheshltterspeed 1-2 sec.In thiscase, is and1mXobjecl areused' ves some types coor fim mayrequne of coor
OM sdesSLRcame6

Easeol opralion

.Manual winding oi 35mm lim .Use of cablefe ease meter Useot exposure (EMM,7)

.Manual windng of filrn .Use of cable release .Use of VarmagniFnder lof rocusng

.tlse ol buill-n meier exposure

Useol large forfial lilrn

needed photomicrography for Equipment

: st checkthe charl to seeif lhe equipment -:.uned ior pholomicrography beencom has :'.iely assembld.fihe lypeslistedbeloware - mostbasicattachmenls needed taking for roto6 ol stained specimens lransmittd in lighl.

. quip49!! !99!ie! !!!!9pgEP!

(m r :ocusiigmagnifier I :ocusingTelescope ! 35mm camera back . iLrlomalicaeosure body . t | ,

:onneciingcord lrgeformal cameraback :dapter for largeJormal camera tulomatic exposlrecontrollnil

df^

-rff

P"
U-

B _slruclion manlals t :crm lor recording data rr leaningutensilsi blower, cleaning iquid, ?1SCeaning lissle

oOQ

ooa

6") t n
U'

Suitable locations for setting themicroscope up

Ressons unsuitable for localionsand corectivemeasures

Ga
T|Ealm6nl .Remove lhe microscope Jromthe soufce of vibralions .Use a slurdy lableas suppod .Use a vibralion prool lable

Unsultabl room

Consqugnces

.Localedtooclose mechanical to appliarices or machinery cancauseexlernal that Mbralions Blurred image a rcsull vibralions as of .P aces in whichthe vibralon ol persons pasl can be transmitled walking .Use of lhe microscope a windo/ near

Brlghl light lromthewndowprevents near correci .Sei up lhe microscop a wall .Position microscope sucha way locusrng lhe in lhatihe overhead iallsin slightty light in froni oi the microscope. with .Place whereroomlighl entersthe eyepiece ljght flares rcproduced the .Cover ihe eyepieces caps or are on .Shui oul siraylighlgetlingntotheeyepiece photo or thefocusing lelescope changing by lhe oplicaPalhseleclor .sel lhe microscope in another .A dusryand dlrly room up room B ackspols reproduced lhe specimen are on .Cover the wholemicroscope a dusf .Placenea.a winclow with where dustcanenier image fromlhe oulside Exampleol a suitable room lor photomlcrography

suitable eyepieces and Objectives photo for photomicrography

Obieciives
:.' pholomicrography cbjechigh'resolulion las wilh flahessall the way10the periphery Oi fieldare requircd. the LB(ongr - visual :a"e ) objectives, seies S PlanApochromal, and a >an Achromai, D PlanAchromal, Plan ._c oi lhe short-barrel objeclives, ,rccchromal PlanAchromal typesare and Corct comblnalions ol obioctiYeand eyplc with FK/P Combination Combination with NFK

tlpto eyepleces

-e pholoeyepiece optically coanpensated is lo E .ermil the objective deliverils full r.'cmance on the film plane.ll musl be wilh the objectives. :..'eclly malched

s PlanApochromat eeris

Shod.bargl Achromal sties

Differencesthe peripheral in images depending objective on design

Accordng lo lhe lype 01objectve the per pheryoi both lhe observed imaqeand the pholographed rnagernayappear ol locus oul This efiecl s causedby lhe pe.formance characlerslics Achromallype oblecives oj Us ng Pan Achromaloblectves however. wi res! t . a sharpand Tlatmageexlendng a lhe way 10the perpheryof the Jeld

iL

y!

rhe rierd ,rarnessor a D Pan Achromai objeclive eiiend nq a th way lo the pefprrery s superof to rhe one orovidedbv a D ach/omal 0b _.civa

Difference resolution in depending on the typeof objective

:- .: -: on n lhe center the lieldoi borh of _- :cserved thepholoqraphed and images : _. : according lhe typeof objectve. to _ - -:c cassobjeclive series, Pan ApoS :-:1s. as wel the S PanAch.omal - :: crovide resolulion. superor

: ; i- Apochromat NFK33XLD 40x.

ligss-ffi
IU*H
Gives srrarp ,esorutor .r lh whoe mase. allowino obseflation oi mlnul dela s

Sill'9oT.--tl?

Somehintsaboutthe objective
It ls importanl lo choos en obictiye sultable ior your spcltlc purpo66 For all objectives, properusedepends lhe on specilicpurpose, sometypesrequiring some adjuslments. orderto tulv rJtiiize ln the you objective, shouldknowthe meaning oi the variousnumbers letiersengraved and on lhe objeclive barrel.
objFljw typ and hagntfication Numicarapenu,e (N.4.)

1. Obictlvewlth conction collsr (N.A.) Method ot adiustnent In objeclives a numeical with apenure above0.6- xcluding oifimmersion l}pes(1)Set scale lhe 0.17mm position, the lo lhe thickness the coverglassslrongly oi ailecls ihe imagequalily. Coverglasslhick(2)Rotate coreclion ihe collar or 3 gradua. 2 nessis theoretically designaled 0.l7mm, al llon marks(0.02{.o3mm) the direction in although actualpraciicethis may vary by in oi 2, and refocus.lf lhe imageis sharper +0.3rnm- oplically By compensaling this ior (1), thanunder rotate collar the anolher thickness devation, correction the collar 2 or 3 graduation maks in the same assures best image. the direclionand tocusagain. (3)lt you are not satisfied wilh the image quality, rotating collarin the opposite try the direction 1-2gradualion tor marks, r+ focusing comparing image. and the (4)Gradually reduce amount rolalionot the oi the correclion collar,andtry to iind the op iimumcondilion repealinq by steps(2H3). r-Ihe abo,/eadjustmenl procedures musl be repeated everyiime lhe specimen is changed.

2. Obieclhrwlth lds dlaphragm Some obiectives abo\,lOX equipped are with prevenls an irisdiaphragm. diaphragm This in direcl lightfrornenteing the objective clarKield lransmitted or lightlluorescence observalion. watching By bothconlrastand resolution the image,lhe apenurecan of posilion. be adjusled the optimum to

la -r at
d

raa aJ al I
I

aa

-a

D Pranf{X, S PlanADo100X, s Pt.n ADo100x Nc

-r

s malk indicares rhat no covr

: \o-covr (NC) objeclive ::cmens wilhoui coverglasssuch as :-: objectvesbearinq mark NC the : :over glassattectsthe imasec arity, .!h ch is padicuarlyobvous in lhe case :.tves with a largenumerca aperlure. ':s!1, when observlng specmens nol : :C by a cover glassat hqh maqnl : specia NC oblecliveis used. . ewingufcoveredspecmensthfough : .bjecllves, on y poor images w th 'ares and insuficieniresoluloncan

'i,,'
'-,
'

) \]ruSi
Li I'

'l /;l- ( )'r

w sp@imen ihouicoverolass NcsPanApoloox

)'. d"
.,.-.

3 ;+:'\)/ \

4. Nocovor lmx dry objeclive High.maqncalionI00Xobleclives uslaly l are oj lhe oll mmersion type bul ior the nocover l00X objeclve a dry type is avalabe Using this objectivelogelherwth other non. imme6on objeclves,e.g. nGcovef40X lhe and 60X.obviates needlo pul mmersion o on llre specmen slide The l00X dry type lens can aso be usedfor pholomc fography. bLrlior achevng oplimumpiclurequaily, objeclve s the lse oi an oil'immerson

-ous LB sries covfobjeclvesi no : rn 40X,NC D P.n FL 60x NC S Pran : : ! a ^ d N CS P a n r m x d r y

SDecimen wlhoul cove,s ass S PlanApo r00X

Obletlve lypeand m.gnific.tlon Numdir aperluc (N.A-) Mehanial llb lnglh ofthenlcD6cop Tnlckns ot lie cdor Olas

'Use only oll specilied by the manutacluret li youusold Cargill or cedar lhe oil oll, obieclivecannotdisplayils lull potentiat, sincetheir difiaclion coetficienls differ irom lhe nominalvalue. oil is llnted. affecls li the it lhe quallyof colorrcndilion colorphotos. in Theretore, is advisable useonlymanufac. il to lurerspecified In parlicular fluorescence oil. for examinalion, onlylhe iluorescenceiree use oil provided thelluorescence wilh microscopes. clicksinlothe lighipalh.Make surelhai lhe objective lronl lensis tully immersed ntotheoil on lhe speclmen slide. (4)lftheirnage notin vieq slowyrolale is lhe linetocusing knob it comes focus. lill into I Makesurethattheobjective fronllensdoes notgel 1oo close the specimen, lo since lhe image deterioraleair bubbtes will il gel inlolhe oil.ll a haze seems coverparls to of lhe image, even lhough is in focus, it swng the nosepiece or 2 timesfrornlhe 1 cljckslopin orderto remove air bubbles. be

rb al -

I l}t all -rl

5. Oil.immersionobjectlv Useof the oil-innercionobiective Oiljmmersion objeclives a numerical have apenureabovel-0 and use manulacturerspeciiied belween lront lensol the oi the (Oil objeclive lhe specimen. mmersiof and objeciives carry lhe mark "oii .) In orderio makefull lse ol the resolving powerof the objeclive,il is preterable useoil also to between condensef lhe front lensandthe

(2)Rolale revolving nosepiece lhal lhe lhe so oilimmersion objective pointing is towards you,andapply io theirontlensol oii

hg

How to apply the oil (l)Focus lhe specimen, the 10X, on wilh or 40Xobiective bring desired and lhe specimen detailin ihe lietd of view

(3)Apply lo the specimen oi surfaceand rotate nosepiece lhe untillheoii.immersiorl objective, tip likewise ils covercdwilh oil,

'

lf the lmagestill doesnol improve, remove the eyepiece illuslraled as above,check ior air bubbles viewing backtens by the of lhe objective, wipoii the oil and reapply oil.
- - 1..j .i t.,,,. t. cte.n an a I.

ObjeclivBtyp. .nd magnltrcatlon NlmsdcalaperllB {N.AJ Mehanicallub. l.ngth ol lh. mlcrosope Thickn6s ol th cd.r glss

'Us only oil spcified th6 by manulaciuler ll youuseold Cargill or cedaroil,the oil objeclivecannoldisplayils fu I polential, sincelheir diffraciioncoeflicienls ditfer lrcm thenominal value. lheoil s tinled, aitecls lf ii the quality colorrendition colorpholos. of n Thereiore, s acfuisable useonlyman!1ac. il to turer-specifed Inpadcular lluorescence oil, for examination, onlythe fluorescenceiree use oil provicled thefluorescence wilh micrGcopes. clicksinlothe lightpalh.Make surelhai lhe objective lronl ens is fully immerced lntotheoil on lhe specirnen slide. (4)lftheirnage notin vieq slowyrctate is lhe linelocusing knob it comes focus. lll into I Makesurcthaltheobjective fronltens does notgel too close the specimen, lo since the image deterioraleair bubbtes will get il inlo the oil. lf a hazeseemsto coverparls of lhe image, even though is in focus, it siwing nosepiece or 2 limesl.om lhe the 1 clickslopin orderto remove air bubbtes. the

:ta
! -

!ta q 'I

(2)Rotale revoving 5. Oiljmmr$on objeclhr the nosepiece thallhe so Useof the oil-imnercion obiective objecliveis pointing towalds oil-immercion you,and appy o I lo lhe lronl lensof Oijmmerslon objeclives a numerical have lhe objeclive. aperlure above andusemanuiaclurer, 1.0 specilied belweenthe iront lensoi the oil (Ollimmersion objective the specimen. and objectives carry the mark 'o l ,) In orderto makelull lse oi th resolving powerol the objeciive,I is preferable useoil also io belween condenser lensandlhe lhe lront

ob.,

How to apply the oll (l)Focus the spcimen, the 1OX, on wilh or 40Xoblective bing the desired and specimen delailin the fieldol view

(3)Apply 10the specimen oil surface and rolate nosepiece the untillheoiljmrnerson objective, lip likewise ils co\redwilh oi,

'

li lhe image slill does not improve, remove lhe eyepiece as illlstrated above, check for air blbb es by viewinglhe back lens ol the objeclive, wipe off the oil and reapplyoil.
.i . ,). I i. ./.;. .i. oii. --t.: ,. a..e 32 l

Selection a condenser of
:rrpcse ol lhe condensers to eficenlly 'le ighl emanalng lrom the lighl ! on lhe specjnren. creale ightn! 1rr i :.is malchng the oblecliveand lhls r. de a be11er image Depend on the ng : . a a b e . P a d c u l a r y i l h L r l r ro w m a g w a .n obleclvessuch as lX 2X and 4X. : n s i k e u n e v e n u m i n a l o n n di n s u l i a f i c e n ta m o ! n lo f r g h l a 1 l h ep e r p h e r y r e a key 1oocclr Therelore make slre lo lse lheseobjeclles n combnalon wtir lhe ! tra ow magnificaton condenser n orde. 1r]oblan betterphotoswtir the S Pan FL 2X objectve.lse oi tire Lr ow tra nragnilicalion condenser BH2ULC s

:nbination of BH2 series condensersand LA obiectives

Condenser

:.r2aAc

:-2 SC

:-2 CA

:-2 ULC

procedures Observation
Now we are Ina y gettng to observalion bul firsl makesure that no dust or d ri s on ihe obleclive, eyepiece, and specimen. lvake t a habt lo check ior dirt beforeyoLl Lrse mcroscope sinced rt prevenls the locusingand resulls n poor imagequaily

ol 1 Placement filters
Turf on lhe ma n swtch and adluslthe voilagelo poston Pholo(ca 9V)

t Adiustment interyupillary of

r,-

r4 dislance

I 3
a

ob .
C

h^
=-1.

tt'@

(3)Place specimen the stageand focus the on wth a lox objective @_

( 1 ) P a c eh e i g h tb a a n c i n gi l l e r L B D . 2 N ) ( l f of the lighl ext wlndow.

(4)Adl!sllhe nterplpilarydslance |]nl boU. leil and righl vew fieds mefge nio one. (2)Pacean ND i ter in the slot close1olhe amp housng Dependlng the objectve on r.agniiication and the densly oi the specmen.Lrse lhis fltef so thal t provides enoughbrghlnessfor easyexa..inaton.

Diopteradlusiment
rc Jsllhe diopler sul lhe observels lo -,.esighl. melhod flersaccording Ihe d 10 I rnlessthe diopieris adjusted, parfocalily ' .r I nol be maintained hen lhe objeclive

centering { Condenser

the : r;_enusing WHKlox eyepieces, .,:-s is adjusled lhe focus knobs with ng .. lhrcughthe .ighl eyepEce ^-e observing aiopleradjuslmeni ing 6) on the letl 'd): s thenadjusted maximum lor image ;'f lor the lefteye.

(8)Alter on with locuslng lhe specimen lhe (7)Alrhattime,adjusl irnage lhalthe so lhe rotatethe lie d slop A in I 0X objective, of crossinesn lhecenter lhelramemask the lhe d rection lhe arrowandreduce of as areclearydslinguished twoseparaie io field irisdiaphragm d ameter a minimum. ines.Thenadjuslthe focusby roiating Then move condenser lop the lrom slowly knob thelinefocuslng so thatthecross the helghl to bonom using condenser by image in Tocus are Inesandlhe specimen knoband stop at a posilion adjuslnrenl I ghl_eye mullaneously. compleiing Alter s ir.age iis where rield diaphgm the lhe lor also adlustmeni, adjusl diopter lhe is sharply deiined. by rolatingsecllon A as n (5). lefl eye mage lhe 1o the I Move fieldirisdiaphragm cenier thevsuallied wilhlhe con' of cenlerlng knobsc . denser

I
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I S ncethe \,pe of the lindereyepiece ditlers (g)Open lieldiis diaphragm accoding lo lhe sizeoJlhe film used, the image lntil choose typesuitable yolr patticular lhe for it almosl louches periphery thevislal the ol iield, makesomelinalcenlering and For adjuslmenls. normal observation make conditions, thediaphlagm slighly EwlrKlox larger lhanlhe visual field. E wFrhlox iris I lfthefield diaphragm cannot sharply be A'- usnga iinder eyepiece pholomicrG E wHkiox4 1 / 4 x 3 1 / 4 " P oa r o i d o tor focused, checkthe thickness the spec| of a is by menslide, Useslides wilh a lhickness t=.y adjuslmentmade rotaling @wNrro{ -: :r B.of lheeyepiece theright al eye. belween and1.2mm. 0.9

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Handling specimens of
toc Make I a habLl eantirespecmenreg! ary bolh beforeand afler obsetualion Jlsl as wlh lenses 1is mosl mportanllo work wilh a c ean specmen Nlakesure thal no dlsl pairces stck lo the specimen when you store I and do not toLrch gass slrlaces the

y H o l dl h e A l a s ss l i d e a l l h e c o r n e r o n , o r h o d l h g a s s s l i d b e l w e e n o u r r i n 0 e . s y

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F i n o e @ a n l s o r h e . o v e rg a s s n

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T h e i m a a ei s b u i i e d a o n s t h e l r a c s o l y o u . I n9elpnnls

Noteson specimenplacementon ihe stage on when you pace a spcimen the staqe the specmen irolderis openedwide and i t s re easedrapdly.i1wl h t the edgeoj the slrdeand damagethe g ass.Aner coni rm ng put n pace has lhatlhe spcimen benlirmly llre sprng loadedspecmen holde...u$ be retracted sowLyso thal il genly lolches lhe n casethe s de is damagedcarefuy renrove tny glassffagmenls.I fragments the are eil on lhe stage.lheymay calse inilres. s or the specimen de may be paced n a t td poston on lhe stage.causng one sde i o i t h e v i s l a i e l d t o b e o L ro i ' c c ! s

o n es d e o i r h ev s u a r { i e ri d b s

Useof the fieldirisdiaphragm

(with35mmJilm) telescope ljsing the Plvll0AD tunet masklocusing . ' e d l r l sd a p h r a g m e v e s1 r r o l u stlh e s a - "ated areaon the specmendependn! '',e obleclveoowe. lh s daohragmpays a r . a r o e d u rn 9 D h o i o n r o l r a p h y n d i c ._:. ode thannecessary uninal n! gh1 r . ecred..d scalteredrre_o! ly on the a : ren eslLlrng a ossoi mageconlrast in : : i g d o w n h el e d d l a p n r a g 1 0l ! s 1 t m :1d the irame rel c e areav/i resu1 n conlrasi : rlreph c mageswih rmproved s stopDe.j do!,/n :re ted rs dapl-rra_om : c ose io the irame retc e tire ohoto :!red ma_oe be cLrt tfrecorners may ai ! d a p h r a g n , s h o Lhd r e T o be o o e n d ir e re trly morelhan 1l.ere:c e slro\ls

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Useof the aperture diaphragm iris andits effeits

The diaphragm rnounled the condenser on s ca ed aperturers diaphragmTheiunclion oi this diaphragm lo maintain mlm is opl condilions image resolulon. oi conlrasland ioca deplhby adlusling nurrerical the apenure of the lLrmnalion syslemdepend on lhe ng nur.ericalaperlLreof the objective use. in For most specimens opllmur. irnagequajly s achreved lhe apenuredaplrragm s il adjusted between lo 60% and 80% ol lhe

0)O rcl viewng wilh lhe eyepice

s
nis rh Reducino apr1ure daprragmlo 6os0%ot lh numeica apenure lhe obleclve. or

How lo adjusl the aperlure ids diaphragm Thereare lwo methods adlushent: oi Pu oul lhe eyepecewith lhe specmen n locus and then adlusllhe daphfagm!,vhlle watchng lhe rs al the rear ioca planeoi the obleclivas n pholo(1):usethe gradLra ton markson tire condenser. in (2) as

(2)Usnq oraduar ma*s on rhe lhe oi Samp e Us nOaloxoblecrive wilh a numercalapduf ol 0.25 andreducing to ll 3 0 % , i h a q G d uo n m a l k ar on lhe connenser should b es e ra r0 2 ( = 0 . 2 s ' 0 3 ) .

I
" *" "*n"* draphrasm I -*o resolLilion irislhe aperture Example Whsn uslng S Plan Apo 20)( NFK 2.5X Fge if deleriorales
I I I I I I I I I I I I I .: d aphraom slopped is downloo much r'h lheexceplion specialy ol slained lhn not .ec rnens diaphragm lhe should be sDped doBnlowerlhan lo 60% ol lhe nrencal apedLire Iheobieclive. ol I Ar effeclsrmrl lo stoppng downthe rDenLrfe diaphragm be achieved rris can :t moving condenser lhe clownward bul :rislends interferewith basic illumina' to lhe :o1 iunclion lhe condenser results of and _ uneven ilumination. always lhe Thus use : aphragm clonot rnove condenser. and lhe

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Fully opnedposlilon

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Conn4l b enhanced. Details also are crany vblble,.nd localdepthls incr6ased, rsulling an oprimum in

g./o Re.olurion d.r.rioralesas a resull

Basic focusing methods

Focusing lor obseruation A lOX obleclve s usedas lhe slandard for locusng.thef the objecrlves changed lrom r(]X lo 4X and rrom rOXto 40X and i!(her lo 100XDo nolchange abrupilyirom iowmag n ficaton (2X and 4X)to h gh (40Xand 100X) As a resLr of lhe imled eye oi the obsetuer t and the largeioca deplhoi low magnilicatan objectves(2X and 4X) lhe ironl mounll)1 the h gh magniticat objectvecouldtouch on the specrrnen surtacewhen the revovng

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AJleriocusng on the specmen $,th lOX obieclivesel the upper mt ol the coarse adllslrnentexcursonwillr the preioclsing Movethe specmendetai 1{)be examlned to the centerof lhe vsua leld and increase maqnilicaton rolalingthe nospece. by

l g iI

Focusing lor pholomicrography Focusng dlring photography done ether s throughthe locusinglelescope the photo of graphlcaltachmenl lhrolgh the eyeoieces or of the binocLr tube.When locusngthroLrgh ar lhe eyepieces. jnder eyepiece a mLrst be used Pror to pholomicrography fnder the eyepecehas 10be foclsed by meansoTi1s f o c u s n gr o n tl e n s . o m a k ec e a r l yv i s b e l t doLrb cross linesas 1wopara el nes ln e lfre cenlerot the lramingrelice

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Focusing lelescope ol lhe photographic For pholoqraphy specimen the locus mLrsl be adllsled wth the same eye wilh wh ch lle .ross ines were foclsed

Finder eyepieceol lhe binocular tube Focusngthroughthe binocLr tube s ar possbrewlh microscopes Vanor BHS.BFT and BtsTU

I
l 3

4r!

s;

1. Focusing when using 1x,4 and 4x obieclives Foclsingerrorsocc!r qu le irequentywth low nragn flcaton oblectves Tierelorelse a l o c u s n gm a g n j e ra n d a d l u $t h e i o c L r s lsled beow by iolow ng the procedures shoud Bui beiorethat the pholographer adjustdioptera1the cross nes oTthe J o c u s n ge e s c o p e r t h e f n d e re y e p e c e i o

2. Focusing when using 10X and 20X rs by FocLrsrng achieved adjustnglhe c r o s sI n e s s o l h a tt h e ya r e c e a r y v i s b l e . knob and then rolatng tire lineJocusng Lrnt both lhe cross nes and lhe mage 0 1t h e s p e cm e na r e c l e a r yv i s b e s m ! laneousy.By sLghly mov ng yolr eyes oi to a sdes. see i the posnLons lhe cross inesand the specmen mage'1onol s h i l f t i r i s s t h e c a s el h e P c 1 ! r es n focLrs t they do sh it. the tocls rs not

'rage is s ghlly oul oi iocls aner l . : o j m a g n l c a l i o n s e o ny t h e f n e _j knobior refocLrsing partcLr n ar rse the coarselocusngknob for _! a h gh-magnlflcat objectve. s nce on . a dangerlhal the obleclve w rLn - specmen ana uer oamageo

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p r o p e r y g n e da n d m l s l a w tl-rtire lne jocusn9 knob 3. Focusing when using ,lox and 10{x Adl!$ the cross nes so tfral lrreyare c eary v sibe. llren s owy adlustihe locLrs imageu71h l ne lhe ot the speclmen locusng knob lnli cross Ines and lmage spec..en are c eary vsDle oi tl-re

i a : : ' e y e p l e c es o r o v d e d w t h o c a tn 9 : . n must be firmy nse(ed nto the r ' t h e e y e p i e cs e e v e f l h e p n l s e _-_t rnserted.correcliocLrsngbecomes : e a n dl h e m a g e w b e o L t o i f o c l s .

Camp the iocusingma!nlfleron the 'oclsing leescope.afd side the top sec readluslLng tocls the ron n or olr thereby is al lhe cross lnes. FocLrs co(ecl t bolh the cross lnes and lhe specmen m a g ea r e c e a r y v s i b e s i m u l a n e o u s y

Basic focusing methods

Focusing lor observalion A lOX obleclve s usedas lhe standard ior locusng.then the objecllves chanled from rOXlo 4X and irom 10Xlo.10Xand ilnher lo I 00X Do noi change abrlpl y iromlow mag nirical0n(2Xand lxl to hrgh{40Xand t00X) As a res! t 01llre lmled eye oi lhe observer and the argeioca depthof ow magnificaton (2X obleclives and 4X] lhe Tronlmountoi the highmagniiicalion obleclive coud louch lhe specimen sLrrlace when lhe revolv ng

3
I

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After ioclsing on lhe specimen with l0X oblectve.set the lpper mil oJ lhe coarse adjustmenl exclrson wth lhe preiocus n9 fvlove specr.fendela to be examned 10 lhe lfre cenlero1the vsLra fied and ncrease maqnlcalion by rolalng the nosepece

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Focusing lor pholomicrography Foclsrngdlrng pholography doneeilher s throughthe iocusng leescopeoi the pholograprr attachmenl tlrrouqh eyepeces c or the o t t h e b n o c u a rl u b e W h e n o c u s f g t h r o l q h l l h e e y e p e c e s . f n d e re y e p e c e u s tb e a m used Priorlo photom crography inder lhe eyepecehas 10 be focLrsed meansof ls by focls ng iront ens to rnakectearty vrsbte d o u b ec r o s sl i n e s s l w o p a r a l e l n e s n a l h e c e n l e r l t h el r a m i n g e l c e o r

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Focusing tlescope ol the photographic For pholography specimen the focusmLrsl b e a d l l s l e d l h t h e s a m ee y ew l h w h i c h w l"e .ross lnes were foclsed

Finder eyepieceol ihe binocular rube Focusnglhroughthe binoc! ar lube s possbrevilh microscopes Vanox.BHS.BNT ard BrITU

fs

1. Focusing when using 1X, 2X, and 4X q F o c u s n g r r o r s c c u r ! i l ej r e q u e n t y w i l h e o Theretote use ow magni cal on obieclives and adlu$ the iocls ng a focLrs magnilier by fo owingthe procedlres sled beow should Blt bejorethal llre pirolographer adlusldopler al the cross Lnesof the tocls ng leescopeor lhe fnder eyepece

2. Focusing when using 10X and 20x Focusng s ach evedby adlustnglhe c r o s sl n e ss o t h a tt h e ya r e c a r y v s b e . the and lhen ro1aling fne ioclsng knob unt borhthe cross ines afd the mage o t l h e s p e c l m e a r ec e a r y v s b l e s i m u n yoLrr eyes taneousy By s ghlly moMng to a sides see I the posilbnsor the lmagedo noi cross lnesand llre specimen shil li th s s lhe case the Pclure s n iocls li they do shjl. the iocus is nol

-')I :,:,'"Hll"l
: male is slghty oul oi jocus after ls t e 0 1m a g n i c a l l o n , e o n y t h e l n e : -! knobior reiocusingIn pancuar : lse the coarselocusng knob for . r g a h g l r . m a gfn a t o no b l e c t v e . n c e c s s a dangerthal the oblecl&elvr rln : specmen and get damaged

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properya gndand musl be readlusled wth lhe fne ioclsing nab k 3. Focusing when using 4OXand 10ox Adilsi the cross nes so thal lhey are c eary v s b e thenslowy adlustlhe locLs imagewlh the r ne ol the specirnen jocusng knob un1 cross lnes and image v o f t h e s p e cm e na r e c l e a r Y s b l e

r: r eyepce s provicied lh a localing w _. pln musl be I rm y inserledinto tl-re ::r y nserled. cotrect locls ngbecomes : o e . a n d l h em a g e w i b e o l t o f i o c u s

on Camp llre iocls ng magnifier lhe IocLrsng ieescope and s de the lop sec the lon n or o!1.ihrebyreadjustng tocLrs at lire cross lnes Focus s correcl 1 boththe cross nes and the specmen i m a g e r e c e a r l yv i s b e s m L r l a n e o L r s l y . a

Howto spotdirtandspecks dustin the of optical systems the microscope of and the photomicrographic attachment
panc es are sor.el mes noted D ( and dLrst dur ng observaliofor phologrpahy. pin but pontingthe r exact localionnraybe diiiicuit Whenphotographing imponantSpec mens which cannolbe photographed again11 can b e r e a l yi r u s l r a l j nigd u s tp a d i c e s r ev s i b e a on tlre pclLrre. eliectivemethod s thereAn fore requred thal w hep you deiecl I Sinced lat ocatons o r_and maycome r n i irom panicies lhe n m lisei (oadng. j m oj transpo().regularchecksard necessary I lf the aperlLrre daphragmis.edlced below 30%. dirl and dusl Specks becornecearly Dlst speckson the pr sm insde the obser' valionlube.on lhe insideof the objeciive and on lhe jnsde oi the rn croscope base c a n n o b e w p d o J le a s y . i c e a n n g t becomes necessary conlactyourOympls

Obser. grapny \frf' oescrirrion

Melhod ol verilication

Rernove adapter large{orrnal the ior cameras

and check ior dirt by peerng ln lhrouglrthe lop li you spot dilr unsc.ewand removellre re ay ens

z 199!li9
prism :91 Camera a

Rotalelhe lop lens eement as you observelhe

Selto,Time mode, open shulterandpeerin lhe

phgto ecein prace, rotate q"lig919l r".9"" 9lep

-_ r path Oplical Tubeenglh correcton objectve Specmen for t andcheck movngdustparlces.
pa cha^ge rfe op_ical f 4- le alernarelyooqetu g lhroughlhe foclsing leescopeor lhe fnder

Eitherremovephotoeyeplece and checkior dusl

O O

Remove observation iromthe mcroscope lhe tube ior body andcheck prisrn lhe su.face fngerpr nts or conlaminalion
ce ovF the obectNp roa l\e nos6prale d o

checkt lor d rt or conlaminalon.

Observe specmen and move lt in lhe fied. lhe li dust is on lhe specmen,il wi also move. Remove condenser lhe lrom the mlcrcscope and look for dlir and oi deposits. Checkthe iilef after removing lrom the m.rG it scooe oase

_O !0 Condenser ; ; *
l? Lghl exil

^ -

. '

Swilch the lluminalion examne lens on and lhe

check lhe irosledg ass Aemovelhe amp houslng and checkrhe enses in lhe colleclorassembly. Bemovethe bulb trom its sockeland check sgns of backenng fingerprnts. dirr etc.

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frame Howto clean microscope the

:ii _son the rnicrocope lramearelirsl Mslerials usd lor th mlcroscooe ..d wltha piece clolhwetedwitha ol i: amounlof neutraldelergent, then and ..a clean wltha piece clolhlhal has of )ir- ,mmerced luke-warm ln water Butmake L-. roi io louchlhe lenssection,rvhile :er_ ng the microscope ifame. \. : using which may organcsolvenls --:ge paslicparls. ::r duslconlam nation adheres thal 10 wipe = _ - pansand is difficultto remove, r: . pieceof clothor soll tissuepaperthat -3: :een soaked a mixlureoi 7 partsether in r: I parlsalcohol, Keep m xture lhe away +:- claslic panslo prevenl damage.

E E E

Howto clean optical the system

'p6ora

s . o . .o e r . o r .- " r o surtace canformay berenroved a t wth

bower Yo! shoLr irowever. d make a habl oi covering the mcroscopewth a dlstprool cover after eacn lse

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Cleaning melhod Tduslspolsonoptca glassessuch lenses as prisms and i ters are e11 unallended. the dLSrbecomes ficLri ro renrole and may d calse mold By alwayskeepngoptca glass s!rfaces clean.yo! avojdma nlenanceprob ems ard prolong lhe ie of yolr mcroscope Clean oj lhe ens suriacesapp es ony 10 ng exposed areasoi obleclives. eyepeces lirlers and condensersIf lnlerna or malorcean ng pease conlactyoLrr becomesnecessary. O ympusN4icroscope er dea

+'.

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)t

and oplical To prevent scratches coalings on ! ass removedin and dlsl lhat stcks 10 the rsuriaceswth an ar gunor b owerbrush

Wrap lhe lens lssLrearoLnda woodenor bambooslck as ustraled.

Requiredlools

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or i A r gLrn bower brlsh 2 c e a n n g m x l u r eo t 7 p a r l se l h e ra n d 3 pai(s acohol.or efs cleannq f! d 3 Ol ps wood sl ck a Sojl qauze.lens 1ss!e can aso be ! Magnryng glass An yepiece lsed n pace ot the magnf e

Whencleanng largeg ass slr{aceson both sdes ot an accessory such as a j tr fod two or three ayersof lens lssle soakedn t h e c e a nn g m x t u r e h o d t h e a c c e s s o ra l y ils edges and wipe lrom the cenierlowards llre perpheryas yo! s owly rotale I

Whenc eanng tl-re suriaces the condens: oi and oJ the iighl exil gass hod a pieceoi y l e n sl s s u e b e t w e e n o l r m d d e a n d i n d e r ingers lod I afd $/rapi1arolnd yoLr ind: ringer Thenhod lhe lissuedownwth yo! lhlmb wlrie vr'rpnq lens sudacescea. llre

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Aftercleanng, breatheghllyon the ens has surlace unli lhe whole surface lurned whle,thencheck whelher hazedis ihe where haze the appears lormly. un Spots d sappears y slowlyare not yet wiped on

:.:. lhe lens by pulilnga smallamounlol ? : .eaning ilud or cleaning mi*ure on the . :'a lenstissue.Dlscardeach ens tissue

lenssurface, wipe When a cleaning large if fromthecenler lowards periplrery lhe

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]- : ean examne lens ng, wlth the slrface ]i irom . -.:t. [rng glass. colorrcfected t- -,_ssurface it ooksuneven, is an ind. - :. :nattherearesl I duslspecks and . " . ewngihrough bonom an eye the oi ,oLr useit as a magntying gass can

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When cleaning argelenswilhlenstissue a yoLr yourllnger, should wipe afound wrapped lhe in Iromlhe center towards periphery a Aways,usea cean pornori circuarrnolion. yourindex rotaie as of the ensilssue yoLr

Howto clean oil-immersion objective an

Clan the oil-immersionobieclive dunng Aner in sh ns obseryal wth an oi on mmersonobjectve.wel a pad oj cottonwoo or a pece oi ens lissuewilh a sma amoLrnt of cean ng mxlure conlanrng7 pans ether and 3 pans acohol,to femovol adherng lo the objectve S nce an oil 1 m w I oilen fronl ens t sholld adhere1olhe obieclive be wipedclean l\/ice after eaclrJSe llse only lactory recommendedimmercion oil and rmoveit atter examination i the o femainson the objectveior a ong p e r o do i t r n e . t w l l h a d e n ( e g c e d a r o ) makng t diiiicLrll removeeven if yoLr to y wipe the ens repeated Tlre lens may be ln d a m a g e d l h e p r o c e s sU s eo n y s p e c i d oi. and afler]]se wlpelhe o irom the mmef sion surlaceof the objectve.keepng t lj you lrequentyuse an oll rlrme|sonobleclive olmay contaminate sLrrjace a the oi dry t/pe oblecl ve *hen you changeoblectves To preventlhe o irom adherngto llre rolalethe nosepiece after obleciivecareiully lowerng lhe slage.so thal the o does nol I lhe mage of a dry lr gh-magnifica|on obleclve appearsiuzzy.check for o thal nriglrthaveadhefed10lhe 1ipoTthe
w pe rhe llonl rens oi llre oblecl've r\1

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

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ry.ola Alle. owernslhe sla0e careru nosepiece crickrh dry obleclNe and

Howto clean specimens

r,:ke il a habl lo cean eachspcimen bolll :r'cre andaflerobservation. OlheMise, dirt -r dusiihal youlailed notice to dlrng riervalon mghtappear llre pholo. on ::- cleaning speclmensonc oth,gauze, the a maybe used wiihoul :, seceo1lenslissue :.aing liqrid.Bll ii ihe conlaminal s on brealhe the spec on men ,-.Lrll lo clean, ri:re wipng il.When ceaning specimen, lhe *,nts to note duringcleaning ' iemoval oi as wellas roulnecleaning of :an b donemorceasily lhe specimen il ; removed lrom the stage. : .ihenusing ceaning mxturcor lens : eaning illid, usea moistened or clolh :?aniag lissue. cerlain to apply Be not :.aessive fluid, il mayseepunderneath as -. cover glass darnage specimen, and lhe --:re areMo lypes spcimens: wiltr oi those a ::.ergassandlhosewlhoulacovergass,
rJ:- as blood smeaG

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cleanlncspimns wilh coverqlass

glassos Cleanlng specimsns with cover ?swhencleaning lefses, wpe ol1lhe :'3 dirlwilha piece lens of lssue ighlly .'ened wilhcleaning mixlure. Because : cannolbe complelely removed with roe repeal wiping unlillheol film Cle6ning spcimens wilhoutcover tElss6s ::-eing to uncovered speclmens cannol a 3edofl. Youcan, however, remove lhe :. Tmelsnglhe specimen 5 to 10 for -.s n a xylene bath. There conta are ners :'::. horizonlal verticairnmerson, and :..or selecllon which of depends on

covrorass cleaninsspecmenswrthout

Section2
Whentaking piclurcs in photomicrography
Instructlons modds PM-1oAD for and Operatlno (with PM-1oADS 36mm camera back). . . . . . . . . . .36-37 . Magnification photographlc of equipment. . . . . . . . . . . .38 Dlfferences re6olution in according the combinations to . of objectives eypieces . . . . . . . . . . . . . , . . . . . . . 36 and (effective Settingof photographic magnlllcation magnification)..................................39 Framiog the specimen of .... - . . . . . . . . . . . . . . . . . . . . . 39 photomicrographs. . . . .,O-41 ... Checking the linished ot . . E x p o s ua d j u s t m e s c h n i q u e.s . . . . . . . . . , . , . 4 2 - 4 3 re t nt . . . . ., ., .44-45 L,se theAElock ot failure... . . . . . . . .,16 Compensating a film'sreclproclty for , , , , . , . . . . . . . . . . . . . 47 Ocoforfilm .............. ',|8 aTypoffilters on a Dlfferences colorrendition In dgpendlng the . of comblnatlon film andlightsource .. .. . . . . . . . .. .,$ a Dlfferences color reproductlon in depending .. on differences colortemDerature. . . . .. . . . . . ., .50 in on a Diffrences color rndition in depending the type ot tilm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .51 film. . .. . . .. .. .- . . . . . . . .. . . . . .... .52 O Purchas color of (Cc) [Jseof colorrompensating filtrs whentakino
color photos a T e s tp h o t o g r a p h y . . . . . . . . . . . . . . . 53 .,...........54

...........56 a B l a c k n d - wfh li m i te l . a c o m o a r i s o nd i f f e r efn ltmb r a n d s. . . . . . . . . . . . . . . . 5 6 of . . . . . . . . . . .57 a Filters. . . . . . . . . . . f d P h o t o g r aw ih y o l a r o i | m . . . . . . . . . . . . . . . . . . . . . . 5 8 p th P

tD

Basicintormation photomicrography in

PM-1OAD for Instructions models Operating (with camera back) andPM-10ADS 35mm
1 Setingthe lilm lormat 2 Load film propody ihe

. r l the or 3 confirm winding tnerirm f

Plsh lhe buton ior 35mm camem.

Loadthe ii m nto the cameraand makesure rhat the end ol the iim does nol protrLrde beyondthe spoo groove OlheMSe tire Jramespacingmay not be Lrn lorm.

Wlnd the fim onlo rlre emptyrakeup spoo and advancei110irame m. I To conflrm that rhe fim s winding be cedain thal the f i m f e w n d n g c r a ns f o l a t n gt l s n o t . k rolale the crank 2 or 3 tmes n the d recton oj lhe dotledarow lo pick up lhe slackol the I m nsde the car.era. and aga n check lhe rewndng crank to coniirmthe wndng of lhe i rn

to 7 Centerthe specimen be , photographed the visuallield in

the and R Fgcus adjust apeiure v msotapnragm

Adjustshullsr speed

X
Movelhe area ol llre specmenyo! wanl 10 pholograph the cenler of the visua ield inlo wilh lhe stagecontros.WhenyoLr use mode PM-1oADS. cenler rhe area10be pholo. graphed n the spot mete.ingseciionoTlhe Focuson lhe specmen and adluslthe condefrser apenureirls diaphragm lhal so sutabe cofirasl s achievedThe aperllre .laphragm s ordinary sel at 60% lo 80% or the objecuve nLrmercaapedure Checklh shunerspeed.Adluslllre speed to between 001 and 05 seconds using by ND i itels

Color phologEPhy to Set lhe colorlemperalure malchthe type lhe of color film used.Measure temperature t'v at a blankareanot covered the specimen. I!,De of fflm Daylbhi film

tldrl Dlal pG! helarclng thn CIn ffllor

t.BD-2N I-BT D T

Tumsi.n lllm
for Srlhespeed lhe lih Lrsed. failurc ior Sel lhe characleristics reciprocity for lhe filmused.

whennot usingthe color Thevollageposilion metercTF is: temperature BHSand 8.5-9.5V.for models 4 or 6V lor modelsBHI and BHTU Elackdnd'white photography a Sel the voltageabove6V Normally, gleen lilteris used. (Fat enphaslzing padicular colors rcler ta page57)

the Focusis adjusted eitherthrough tocuslng or the oi telescope the PMI0AD/PM_I0ADS binocular tubeof the microscq*,(rcfer to pages 2627) oADS/ telescope lhe PM_1 of I Thefocusing phololrames includes different PM-1oAD lor diflerentlypesof film. \?tes wilh I l}le lype oJJinder eyepiece iilm sizeand eachtype of nnderelepiece inclucles severalpholoframesindicating oi diilenl magnilicalions NFKphoto is aesure adjustment basedon the lolal lo ion oi the specirnen.freler butlon.After lhe Press shutlerrelease checkior the sound exposure, compleling of ilm wnding.

qt

Magnificationthephotographic equipment of
Photomagnlcalon on the flm surfacevaries wth the prolectonlenglhoi lhe photographc pmentand lhe type of pholoeyeoi-oce eqLr lsed ln a casespirolorna_on fcaton s c o m p u t eb y m u t p y n ! t h eo b j e c t v e a g d m n I cat on andtheprrotoeyepiece magniTicat on v r t h t h e c o e i i ce n t s l e di n t h e t a b e o n rne flgnr

errotogaptricequipment

Photoevepiece

FKNFK{ype

P.type

PM.lOAD P]\,4.1OADS P[.4 10M PN,l,6 BH2 PM 6

0 5x 0.5x 09x 08x 0 4x

Whef usnq modelsPf.'ll0ADor PNrl]OADSrOXobleclive and 2.5XNFKpholoeyepce P h o t o a n1 c a lo n = r 0 x 2 . 5= 2 5 m

Differences resolution to in according the combinations objectives eyepieces and of

Eventholglrlvhenlhe overalpholo magni ca Example:Pholo magnilicalion ol 100X r ! l o n s l h e s a m e . e s o r l 0 na r e s d e p e n d n g L on the combnalon of obleclveand pholo T l e y e p e c e h er e s o u o n o n t h e1 m p a n e s mprovedj lhe magnflcaton oi the photo eyepece s ow and thal oi the objeclve h glr tl [ . As the photoshows.a combnalon of oblec. l a Uve40X and NFK2 5X provdesa c ear dspay of evenminLle magedela Foca becomes slra ow deplh.lrolrever. t

l..

i i '

'

at'

, t.
I

?1 'tr'-t .,...1 lt
1r ' z l{ :'' ..tt... t-

,'l?

' ;"

' o

.''. a r l
t
.e ' .o
', 1

't
. t t
tt

.."tr

.,.a\ I
Oble.l ve S Pan 20X and prrolo eyep ece NFK 5x

^1

,1'.''.

,"- --.'

".

Obrecrve s Pran40X and pholo eyepece NFK 25r

Setting photographic of magnification (effective magnification)

n'en taking pctures, have Jirst you to decide ar rhedesired enlarqemenl Themag. ralio. r_calion ihe film paneis generaly on :E:ermined objective by magnilicalion limes 9oro eyepiece magniiicaiion. the eliec But Si6ctiYe .agnilication *Theetfeclve changes llve magnilicallonthe piclure ol magnilicalionbased the is on wilh the nume cal apenureol the objective, assumpllon ihe picturcis viewedwilhin thal atlording dislinclvision, and ls normally basedon lhe lollowingrealhe closesldistance The values istedbelowdo not applyllten tionship, which mustbe lakenintoaccolnl when enlarging pholomicrcgraph. tryingto proiecl35mrnlormal slides. lhe
5@N.4.<M<1qnN.A. N.A =nune,iczta\dup at th. obta.iE M

tx
S Plan 0.13 65- 130 S Plan 0.16

't0x
S Plan 0.30 S Plan 0.40

mx
S Plan 0.46 230-Affi S Plan o.7D
S Plan

40x
S Plan 0.95

'tmx
S Plan '|.25 S Plan 1.30

tF ol ai.tclive t-4"
EHive atnilication

0.70

-4m 80- 160 1503m 200

350- 700 350-700 475- 950 625 1250 650- r 300

Framing thespecimen of
-! pholography,lhe specimen ollen

becomes morc operation lhe microscope ot of the .rollil inlolhe ohotoframe.Thisoroblem d ficull.Herc, iraming the specimen is carriedout the pferrcd way,by rotating !e overcome by rotaling the camera the slage. in relalion the specimen, lo though

'nE

photomicrographs of Checking the finished

pt|otc Phobrnicrography inuol!sscientjfic gradry,which makesit impolatn/e lhe thal dDlrapher accuralelyrecor(bhis fndings on film.Addliooally, pholograph lhe shorid convya stror estrlelicimpressim lhe to vis/ver Valuable recordsshdid mt be documeitedwith runof-theflill dtolo6. Tofurlher inpro/e your rcsultsin photc micrography. h important you aler4/s lt lhal checkyo|lr o n pholo6.By relerdng the to follo/ving checl@inF,liou cn pinpoinl any problenE conceming Tlln prc liour photo6. cedlo the secondhalf of this section lilled Photomicrcgraphy Techniques. 3. Bfackrndwhlb photogt $if teler to 1. Ouosdonarlallng to all phoiography pages 5557) (l)ls lhe imagein focw f/eter to pagesb-27) photograph (])Dosthe tinished sfrori, and lhe e&o6ureprcperly adjusted? graded (refet to pages 4246) blackard whiletones wilhoul (aHave din or dust speckson the spcimsn areas sivelydark(solidblack)shadow washed{ur(corndelely $/hite) highlights? foundtheirway onlo the photograph? telet to pages2&33) (3)lslhe specimen properly stained? (4)lsthereuneven illuminalion? ffetef fo pages225) Z Coforphologr.phy frefer to pages 47.54) (1)lsthe baclloround (emptyspace) whiteor ol lightgrcy lone? (4ls lhe color of he specimen accuralely reprcduced?

25 nrmKodachrome

1984Dec.l

Purpos

a a
3 6 7 8 I 10 tl 12 13 t4

11

0 17 7
lq
18 18 21

rs 19

ry
i 4

a a_ n.
a 4 a
I 31 3

.v
5

Photomicrography techniques

techniques Exposure adjustment

Whentakng pholom crcgraphs, exposure compensalon necessary is depending ihe on dislibUlon the specimen lhe iield. of in Butii the specimen evenly buted is distr 1. PM.lOAD (60% Averagemeierlng wlth modl PM.lOAD) or within nlegrated spoimeieingrcnge, the is red no compensalionrequ (incaseoi 1X).

ZP at al

Z PM.IOADS rModl can PM.1oADS b u3dtor inleg6ted melering ol 3ool"anctspol melenng ot 1oh.\

Measuring alaa

Soecimencondlllon wlthin ara the l% meledng

dotted Brightf background ield dense spcimens wilhfairly

0.25X

con' ield Brighlf background scattered specrmens laining

0.5x

is d Specimen evenLy slribLned area. wlhin lhe I % metering

1X

Sellhereclpro

lilmbelngused. Aboul (50%) the half 01 wiihln specimen distribuled is the da background

*t

(250lo)of Aboui onefourlh lhe specimen distributed the ls n darkbackground

is Thedarkbackgrcund dotled the lSo/ASA dial sensitivily

Useof theAE lock

By prcssing buttonduringthe automatic AE lock operetion this you exposure rnode, can adjustlhe exposure g,%, P -10lDSfnr6gratd metoring lime indicaled the displaypanet(expected on spot mstering l%) exposure lime, actualexposure time). _lhe Wlen pressing AE 'ock hJtton,the wuning light abo/e it nashe6 indicate to AE lock mo(b. The unil is oow readyfor ptDlogphirE.Tocancelthis tunctton pressthe AE lock buttonagain.This sl|itcheslhe lightoft andthe de\ricere verlsto nonnal autornatic oocure anode. Plr.loAD (averlge mei.riig d}%) Aller poGilior lhe specimen be ng lo photogmphed pressing AE lock and the button,lhe $/amirulightabo/e it tlashes to indicate lock mode.Then,after AE laling the frst photograph. eecr.ire lirne is locked.in. pr6s Tocancel tunclion thjs lheAEbck butlonagain.Thisswitches waming the lightoff, and lhe devicerevertslo the norml automallc e4osure mo(b.

gimlnaling density yadatlooeby AE lock 1. Takingpanoramlcpictues uslng ih AE lock pictureswheieany number For panorama oi copiesis lakenfiom differenlseclions the oi samespecimen whereseveralpholosare or patched logeiher,pertectpanoramic photc graphswilh uniformdensitycan be lakenby locking-in lixed exposure all the pholo a for PanoEmaphotography

z Erampleol uslng lhe AE lock on model PM.'IOADS 4)ln conbination with spot neteing ' specirnen areasrequiring spoi metering of arenol cenlered because faminqprob in ms,lheyarefirslposilioned the cenler Thenthe andthe AE lock is activated. soecimens moved are backlo lheirorlginal aosilion pholographed. and

1 Thspermndetaills al th6 locallon bul whereit is to be pholographed, cannolbe msuGd exposure

delailto 2 Aller movhqth spocinen areaand aclival,ng the measuring dtailcan the aE lock,lhespclmn now be movedbacklo iis o.loinal postllon s and rh6 pholooraph lakn

2)!!9!9gEP!!!9vt!!i! lJ
to naximum exposurctime ' exposure ls display c oselo the maxrmum fluores_ during eroosure lime,lor example the :ence photomicrography, pholographic an mighldisplay underexposure eouipment This exposure. can ^?rning during automalic the by . prevented using AE lock,resulting . ong rme exposure.

ThsAFEry lampcomerro rd .nd warning sounds. an audlble

Compensatinga film's for reciprocity failure

W:thnormally usedpholraphicemutsions Exrnd.: Dai. ior rhe afirraclr&llcs ol rclprodty law teilu|e {Kodrkcotortflm DKD-|/|1) thereis a rule(the reciprocily law)thal determines luminance he lightstriking the of the lilm surfaceAccording this rute,lhe to tolal amountof e&osure is delinedas the produclof the lumimnceand lhe exposure time. For example, anoont of oocure the wita 1,60sec. o@surc at E is the iame as lor I B0 sec,at f11. &Jtfor longer expoGure limesthis ruleno longerapplies, leading to uncler exposure changes color re and in producljon, phenomenon knq n as This is reciprocily lailure_ sjnce,in d|oto law Bul exposure. lJneven color reprcdrction muat compensation lilter are necessary. by For mrcrography, be expoGure compensation for cannot 'll|ecompensatd with a CCflter. additiomldah on recipnccity tailure law be carriedout via the aperlufediaphraqm, abore chan [sts lhe compensation data characterislics conlactthe lilm manufacturcr exposure time is lenglhened shonened or lor rcciprocity failurecharacterislics law when lo obtaina suitable expmurelevel.lt the usingKodachrome ilm lor geneElptrolo 25 reciprocity is set on rnodets dial PM-1OADS micrography. chan showslhat for tong The and PMIoAD, compensaiion carried is expo6ure timesexceeding sec.,bottl 1 oul automatically, resulling pfoper in exDo6ure @rnoensation color time and

Ex8mple comprcallon ol (u,t|n Ustng Fulhhrome ROl00)

No corn. pnselion

Compeft sallon

Colorphotomicrography

Color film
'i.v different typesoi coor i m are oiiered :.e markettoday eavnu tfre Lrser a 2t :: rs 1')lrhich brandto use tor photomrcro :rhy Normaly. day ghl typ '- an ISO/ASA speedof 50.100 s lsed req! re a lghl baancing e microscopes ' i r L B D - 2 .N ) 2

EKachrorne 64 EKachrame 100

Day typ ight

Agfachror.e 18 CT Agiachrome Type 50 S Agiachrome 100 Fulchrome 50D Fujichronre r00D

Eldachrome 50 Agfachrome lype.L 50

t4

::quiremenls forthe selectionof colorlilm I m w t h h q h r e s o l v f g o w e rs f c e p h o l o p . _ .rograplryrequres disoiay dela ed oi -_! d i s p a ym u s tb e a b e t o d s c r m n a t e :.lween the f ne coor dilferentalonscon. : .i e d i n a s n ! e s o e c m e n . : : l h l L rr e p r o d u c t oo i s p e c m e n o o r s n c inoul backgrolndd scooraiion

l. vew or theselactors the iolowng condi tons can be sel ior pholomcrolraohy

! l!!qq1!

(2)Good colorcontrasl
For noma photom crographyn lrans. m l r e d l g hb r g h l f i e d u m i n a l i oc o l o ri m r n wth an lSo/ASA soeed belween and 100wi 50 qla For specal cases ass!re salisiaclorv i1y. s J c ha s p r n t e dp i r bc a t o no r e n l a r g e m e n i s <cd:chro'.e fim oijers lood qla ly ng r.,/.e. oholograprr dark specmens(phase

l?lqlgqJ9!9r !9!!Iro!

conlrasl poarzd ght. 1l!orescence) tirere exmsurelime ncreasesAtiroLrgh r .nay be ercessve gra n for cases reqLr n-o iasl shullersoeedslse flm willr a h gh rSO/ASA speed

Types filters of
Fillrs Seection filers lor usein phoiomicrc oi graphy based lhe tlpe ot I m used. is on lor UseLBD-2N daylight andLBTI lters lilms lor tungsten lilm Forchanging type lighl inlensity, of a neutral use densiiy iiler (ND) is recommended. number the lllter The on rirngives transmssion lhe valle In case oi iillerND6, 94% of rherotaliluminaling lighiis absofbed ony 6% of lhe ighl and is lrafsmiired. Types lilter and their lunctions of F tersior lse n coor pholomicrography comprise io lowing ihe maintypes: lilter I Lighl balancing Toconverl colorlemperalure ihe mlcro lhe oi lghl sorirce the lih being scope to used.

density) lilter eND (neuiral

compensaling) 3 CC (color liltor suchas slghldi Forcolorrendlonproblems pholos. lerences co or hueor ladingin developed in

.F

a
To feduce the lighl inlens ly without atiecllng color lempelalure, when llluminalion too b.ighl. is

Didymium lilter (marked FF a4on Olynpusmod6ls)

Toeriphas color, wiren ze i.e. trying compensate 1o tor the insullicient inlinsicredcolorrendition oi a llm or lo enhance redcolorofthe specimen the or lhe coiorcontrasl Polaroid ol coiorfim.

Heatabsobinglilter

To absorbheal raysemanating trom lhe jghl sourceof lhe m croscope prevent lo damage to or deslruction livespecimers. oi Since filer lhis lransmils smalqlanlty of bluelighi,I may a lo becomenecessary usea color.compensal ng lilter (CCl0Y or CCl0M)\rheniaklngco or photos.

depending in Differences colorrendition of on thecombination filmandlightsource

-.F are tro types0i coor i n's d.y 9r1 sm .r _vrh.n !s n,othe llngslen lype I m - 1 b ra n c e d o r s l n g r i l : n d i L r r l s l e f t o l t c o o r sa b ! s i r . l ' o t o qa t t s o b t a l ^ e o The ta! I r Lrclrc:ses s ta Lrr. 10lse I : . 1 b a a n c e c o ' a r t i c l a g h 1 ). h es e e c lo n :he pr..e- I r. .r r:lcf .he ,ori sc! cF . . i d n g u p o ni h c L l l n ls o ] ' c e V r l l a y . s e r s a v e r p e r e n c eic ea s a c p o n l m c n i h e h r e r o h o t o s p p e a r . !? l c e s s l e y r e d a _ ' ! . l h e yl s e d d a y Slr_ m lnder n.andes

lo or rendilion dependingon the combination ol lilm and lighl source Daylishl.lype(sunlighl) (adificial lighl) Tungsten.type

in reproduction depending Differencescolor in on differences colortemperature

Color lmpralure Dillerencs color reproduclion in dpending dillerences colorlempefature on in Coor temperature designates propenles the of the ight source.A blackbody radator. whenhealed. emanalesighl of dfferenlcoor dependrng the temperature on The properlies oflhe ightsource can be nd caledby reiering to lhe lemperalLrre the backbodyat a fxed oi ter.peratureevel.Ttiis ieature s ca ed coor tenrperatLrre numericaLrnlbeingex the pressedether rn absoute lemperalure or in degreesKevin (K) C o o ri l m f o r u s e n p h o l o m c r o q r a p h y s normaiydaylghl lype bul sincethe ight sourceoi lhe mcroscopeis a tungslen lype ls coo. temperature low (2800.3400rc s and as such il is unsuilable daylighlfllril ior wlh ts color lemperalure 550C6000K oi In orderto achleve propercoor rendilion the a ight baanc ng i ler is used,and llre 10 lungslen ghl s convened dayight

'

>

j , ',.

' shows redsh11 and .doi renoeralure h qh l exhbils a s a'blue s lli color temperalur".low lhe photo s

in depending Differencescolorrendition on the typeof film

for Colorrr'dilion diflerenttypesot film f.om lhs sammanufacfurcr lendsto vary,ard evenditferent orcduclion of lhe same lots typeshov/sljghtdtferences. Whenyou slan to lake color photo6, lirD drouldflrst d6l6mlnelhe oFffmum condilibns to thenlmthrough photogkphs, tst matchlng flem wih boh micrGcooeand soecimen (Fot the methoclol taking t$t photos rctet to page 54) Offlrnce3 color davalopftGit In &pddlr! dr lh. m. ol lllm I}re 6lx ptroloG the rlghtweretaknwllh on flms ol dlflerit brancls, wflfi thE xcp and tbn of lhe ISOASA speedall drotographic conditions wereidentical. vample lhis dearly *lo that properties suchas color rerditior! contrasl, cladtyof baclgrerd, etc.are alldlfof ftrcnt because lhe variousbrards of tilm. Photognphlc condldort BHS, PM.ioAD, LBD,{ (color tompontu|l 5500f0,LBT (color tampor.ture 3,mr+ duii.. 3ped0.1+l sc D.tlglrt tyF

llrngd.r lpe

Purchase colorfilm of
1. Whatlo walchlorwhnbuylng colorlilm I Coorlilm is a hghy sensitive subslance, andenvircnmenta laclors suchas heal andhumdity easily cause changes film n speed coor rendition. and When buyng coor film,avoid camera siores which in the sheves slorng color t m are exposed to sunlghl. I Choose cobr r m wiiha sufficienly only longpeiod before explraiion the dare as marked lhe package. on I Coor fendition lhe samelilm tlpe may oi prodLrction ii diiier, il cornprises dltierenl lots.li youlrequently coor film,you lse photographic canuseil under ideniical conditions avod varaiions thecoor and in ol the t lrn if youbuy argequanlilies irom the sameproduction lol. 3. Poinlscalllngtor spclalattntlon 2. How to slore lilm pinciples composl and whenusinglhe lllm Forthe basic ol on ihe longer colorrendition check lechnicai the literalure I Donotleave Jimin ihe camera on lhe subjecl. hasbeenpoinled As out, perforrnance changes according the con. 1o which f m is slored, dilions under the I Color m is normallykepttherelrigerator n lo protectit irorn the etlecls ol heatand humidjty. Removelhe trom reirgeralor lilm lhe t hourbefore andalor t 10reach room lse lempelure. nol remove fim lrcm Do ihe lhe rc immediatey takinq out ol the after it ,

EE: ;:!

reirigeralor prevenl lo condensalion lhe on

@M

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

I Develop f exposed m as soonas possible.

z
o

160 2/54

;:E
ii
I Even though olherpholograph c condilions colorrendilion vary may maybe idenlical, wlh lowor highambient temperalures. I Do nol usethe fim in a gaseous en(such lorman),sincecolor vironment as rcndilion be aclversely wil aflecled.

sg:
im of Yalldlry(ddEnon dal)

(CC) Useof color-compensating filters whentaking colorphotos

lilters colorcompensaiing arehighly S:nce do sensilive heai, nol usethemclose lo

:c lhe lighl source.

ICC filter is the abbrevialion colorfor filer, compensaling andlhe coloroi ihe parllcular Jilteris markedin capita letters on its rim.Sixdillerenlcolorsare available, eachcolor beingofferedin six dilferenl oi No. degrees densily. 5 andNo.10 are mostwidelyusedin pholomicrography. lcolor fltersare madeby severananu'lacturers. yourOlwpus dealer or Check camera sloreior avaiiability. a repuiable

Color to b rducd BLUE

Colormpn6ating lllt.r rcqulrd

ccY
Red Magenla Blue CYan Green
CCG

CYAN
GNEEN

YELLOW
RED MAGENTA

ccB ccM ccB

Testphotography
1. The need lor lest pholography equ Evenii you |]seaulomatcpholographic p' menl and set bothlhe ISO/ASA speedand lhe rec proclly aw iaiure characteislcs, you cannolbe sure that yo! wil always ior ach eve the perfeclexposLrre everytype ol specimenThe reasonior llrls s that phoiograplrlc equpmenl s mafufactu.ed to malch dstfbLrtonstate,coor and exposure oi ln slandard averagespeclmens. orderto you shold I rst lake obralnbelter resulls, oi tesl pholographs match lhe conditions lo iilers etc the spcimen, fim, nricroscope. 2. Method lor test photography Charl2: Datacharl (when using nadels BHS and PM-12AD Color Colorcom. and reversal filn) lemperelure Exposure (1)Prepare rol oi illm (36 exposlres)and one 1 a irequentyusedspecmen 0.8x No com' (Z)Set lest condilions (seechan 1) and lhe 2 1X pensar on preparea data chait (seecharl 2). Listng 1.25X a the condtons gven n charl 1 wil resLrll in a dala chan 2. TakeyoLr photosbased 0.8x_ 1 bgooK on this chair and delermnelhe optmum 1X 5 o r CC5G pholograph conditions the basjsol lhe c on 10v resulisoblaned on lhe developed m. I a 1.25X

H
it
7 8 9

cc5M

0.8x lx

1,?sI
0.8x 'lx
'| 25X

J9 ssoox sv 1'
Charl l: Testconditions
Measuingwilh CTR No CTR

170mired,180m red, lmmred, 5900K 5500K 5260K Volase 10V, 8V 9V 0.8x, 1.25X

ng Color compensatlon Coorcompensal I lter

3. Evaluatlnglhe test photos observalon oi reversal m transparenc i es can be caried oui eillrerwilh a lighl box or a slideprojeclor, ihe dispiay cotor bLrt of vares wde y wilh lhe colortemperalure ot lheselighisourcesAs a slandard evauaf ior ing the pfopef coor Lrse ghl box wilh a a color lemperalure closeto 5000K. 'Tl]e io lowinglps ol lghi boxesare avaiableon lhe market C Fuji film color box 5000 O Dlrotesl ColorClasse.50 O GeneralEecrric Chroma50 4. Organizingphotographingdata [.4akeI a habil1o recordal dala rearing io bollr lesl phoiography reguarphotc and graphy.lf the pholographic condilions lhat resulledin goodpholosare relaned on i e. lrolbeshoolng and correclon of probems can be caried oul quicklyand efiiclenly.

Cc5Gnocorapensat c,cs\,tr on,

5. Slodng developd film Whenstorng I m ior a ong perod oi tme. precaulon s proteclion the most imporlant frornlight Siice mold is prevalent hot anc in humd paces, t is advlsable keeprhe iim lo liglrty seaed, together wilh a desccant We recamnend storing film in an envnon. nentwith a relative hunidiry of 1U0% and an ambient tenperaturebelow 21oC(71'f ). (fron Kodak Colot OKP 141instructions).

photography Black.and-white

film Black-and-white
crcr : nce the condtons diier ior plrotom phoiography. s i1 : aphyand generaL :cessaryto seecl lhe properiim Joreach '.oe oi pholography photom rogfaphy. In c ! h c o n l r a sjl m w t h r n e g r an i s u s e di n ol -.der to documentm nule slrirclures : oo! ca specmensand to achievesharp :_ololraphic reproduclon. Examples lhs oi 25 n typeare KodakPanatom X. Agiapan c .rd Pan F. and Fll NeopanF

,:jl r#
Jse ol black-and.while liln ':re are manylypesof blackand.wh ilm le j the keytoobla ninggoodpholographs es r e e c t n gt h e r i g h tf l m l o r a p a r l c u l ajro b

Specinren Normaor hgh conkasl oi ior tlreslanedsecllon 25 generapathoogical Agfapan

SO/ASA

32 50 25 32

on the coor 01lhe

Pan KodakTechnical palno09 ca speclmens. 2415 as we as thoseol other Agfaonho25 areas.Wlrenshape s more mpo(anl llran

Srce the conlrastoi the film is very lrigh,the rangeor Pro 64 per exposure very narrcw is expos!reis sel wlhln Uriless 100 +l/3 slep oi lhe opi murn 25 lhe pholow I be ether over

32

Agiapan 400 Dark specmens.and when ong srrltter speed l l o r d H P 5

400 want10shol(enexposlretme e 5 G 1 6 0 0a 1 l h ee x p e n s o f f l m g l a n 400

ryp"" vo, X33 u""t ese *r,"n

Comparison different brands film of

Types fim orovdinga wde gradalon iom oi shadowro h 9lr _ohts featurnggood and conlrastare Fll NeopanF and Kodak PanalomcX. TechnicaPan oiiers s ghty h gher contrasl wh le N4 Ci]pyerimnales n NeopanSS and Tr X on lrre olher hand. havefanly o!\rconliast

Kodak Techncar Pan 24t5

Filters
Ellects ot verlous conlrasl lillers Us ot contrast lillors fllers are usedio conlrollhe conlrast Conirast pholos. or b ack.and'white green Forblack whilepholomicrography, and i tersare normally used, choceoi lhe bul mosleffective depends lhe tlpe of flter on Filterc enhafce colors lhat the specimen. andconlrast the specimen of are: Color ol lh6

Colorol rh tllt r

Green Blue Yellow/orange Oranqe Blue use oi Forreducing conlrasi, a Uller lhesarne men. .oloras lhe spec

for Reasons using a grn lill,ar -iere why aretwo reasons conlrastimproves shef a green llter is used. T)S objective abe(ationsare rnosl nce compensaled lhe green near effectively wavelenglh, oi image arityduelo loss c chromatic abefialionis avenedby a green I ller. 2)Dyes suchas hematoxyln eosin and green in absorb lighlwell,resulting higher lllleris used. conlrast whena green

the red po.lion is nhanced

is lhs rd Porlion reduced

Photography Polaroid@ with film

Oneof the aclntages Polaro litm is thal of d I canbe quick]y vlewed a iinished pholo, as bul il is seldompossible oblaincotor 10 reproducllon comparable thal oi 35mm to 2. Photogrrphlc tchnlqus B i l1J Iackand -wh te photogh phy Usea greenfilter to obtaingoodcontrast. (2)Colotphotogftphy Since overall lhe colorhuetends be either to ighlgreen blue,useCC10-20M or or CC10-20Y compensation. wanl lor ll you io enhance colorconlrast, the FFiiller use available tromyou. Olympus dealer. SincePolaroid is morc easityatfected iilm by reciprocily Jailure law than normaliilm, exposurc lime should adjusted belween be 10 0-05 and0.5seconds.

ol lllm usd ln

r.Pny
,l'x5" (actualda gdnx11.5cm) ISO/ASA speed
4(J0

t 1E' xS th' (actuel slzo73crnx 9.5cn)

I k I a n I Type
107

rack tso/AsA containin! Tlpe speo 8 photos

Sheel

rack coolaining I photos

67

3m0 3000
50

o o
o

52

o
o o o o o o

30m

4m
(P nU negative)
80 80 80
55

665 (PdnU I negalave) 108 I 668 669

50

o o o

58 59 559

80 80

80

14'x5'

filmhoLclers according lhe typeot titm diller to

sh@i lllm-:(s typei m hotdr

Packed fr/m-550 typ tllm hotdef

4500K-LBE!2N cororlemoeraiure olus FF tllto.

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

SectionS
Howto obtalngoodprinis
p . . @-61 . c H o wt o o b t a i n o o d o l o r r i n t s . . . . . . . . . . . o . . . . . . . . . . . . .. . . 62 aPreparingblacLand-white brlnts Maiks on the Dhoto. . . . . . . . . . . . . . . , . . . . . . . . . . . . . . .63 ... during development ......64-65 Howto avoid marks

prints good Howto obtain color

ThelollowlngdiagEmshowshow to get good color pdnts {positiyepdnb). Color negalivelilm

ln pholomlcrography. lhe colorreprodlction oi lin shedpr nts takenwllh negal fi m ve differsirom the observed color-a lact many y usersare probab very muchawa.eof. reasons this thalthetechnician for are Possible who made pinis d d notviewthe speci lhe mencolorlhrough microscope thus a and didnol know actual lhe color, the pholc oa gapher madea mistake whenhe lsed a f ller andlailed notice lhe iilm quality to ihal deteriorated.ordeflo gel beter resulls, In you should lirst photograph specime. lhe tromwhichyouwanl lo get prinlson reversa R6wtsalfllm photo cojor llm, andatlach iinished lhe as reterence sample the negative Pinls 1o lilm. canalsobe made lromcolorrcvercal {ilm, bul lhe process doesnot matchthe pint qualityas obtained from negative lilm.

Color pdnl with Poor color rproduclion

Color pdnt with good color reprcduclion

Preparing prints black-and-white

There many are pubtications ihe technicat on markelllrat expta in delaithowto devetop n iilm. li you planto do yourowr iitm pro cessrng, lhesebooksas reference. use Butsince conlrast photomlcrog.aphy in is ower than withnormal phoiographlc subjecls, youshorild panicular pay attention under. lo developmenl. makea positive ll you print iromanunderdeveloped, /ow-density negalive, an evengrealer reduction contrasl in witl occur, badnglo a poorresuti. Gradeno ol photographlcpapr

1. How to obtain a good negative (])ljse a devetoper malches iitm thal lhe (2)Use developer the allhe lemperalure specilied the manuiacturer by (3)Pdhere the speciiied lo deveopment iime lhal malches sensil llm vity (4)Agilate genlly freqlenlly eliminaie and to uneven oeveopmeni surelhal lhereis nelther much loo i5)Make nor loo Itl e it ng (6)Follow paperspecificaliolswhen the washing prinl the ihe carelutty pfolect and OHandle negative 1 lromfingerp|nts, scraiches dusl and 2. Selectionof phorogEphlc papr pints, youcanvarytlte contrasl Whenmaking depend on lhe t 9 of pinting paper. ng selectthe lpe that bestsuitsbolhthe density thenegative spctmen of and contrast. Theexamples ihe fightshowprinls on made iromlhe samenegative whenusing difierent tlpes 01pinling paper.

Grade nomalcontrast 3

Marks the photo on

dveloFnent 2 Ma*s occuriog dudngprinl devglopment 1.Ma*s as a esult ol negaliv Marks the printtendto slandoul more on prominenllypholomicrography on in than nomal pholosol popleand landscapes. pholomicrcgraphy empoys with lilm Since faiiy highconlrasl, even rosl minute the dif ferences brighlness in showup in the piclure. MarksresuI ng from improper development This deals arealsoveryconspicuous. seclion wth marks resulting developrnenl. lrom

Bothuppd and lowerponbns of lho photoare bnohr* thanlhe centralsection.

Markswith iregular qhlness.howup. b

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

Carelully producl daleandlre. note on quency useof the deveLoper ol (2)Agilale developer y Lrse. How to avoidcrcatingna*s the lhoroughbeiore (1)Agilale developr lhoroughly. lhe (3)Use developer lhe specified the al (2)Keep lemperalure lhe developer the of at 20'C.or as speciiied the manuiaclurer. by (4)Donot workwlh an exposirelhat resllts marks easily can occurif deve or ina developmenl of Less 5 mlnules (3)Since lime lhan (lheresull be exlremely wil clesensitized menttime s too short,set the exposure al that of lheenarger sucha evel develoP developmenl). menttirnes between min30 secand I (s)Agilale baihthoroughly during deve oF lhe 2 min. (4)Agitate balhthoroughly the during develotr (6)Mainiain speciiied lhe lixng lme. y. f)Washlhe filmthorough (s)Conl to agilale nue lhorclghyevenwhile and stopping iixing. (6)Wash pinl lhorcughly runnlng the in waler. paper 300 min Regularbase paper- min Resin-coaled 4.5

processor develop It youusean aLnomatic to paper marks occlr,but will lhe prinling no slncemar!caneasiry / up when sho deveopingin a ty, rhe io lowingpoinls

Howto avoid development marks during

DoYslopment tllm ol
on when Sncema.ks lhe lilm occureasily uslng ree-tlpelank,developmeni a should be donein lhe io lowing maaner the n

t,

'@@@

lmmerse ree in lhe developer, aiter and lhe ii lap 2 turning 2'3 limes, il anolher 3 limes agansl lhe botlom thelankto remove ol a r bubbles sllcknglo the fim surface. Complete process n 56 seconds. this wilh

and Nowremove reellromlhe developer the immerse again. il Aflerrcpeal ng mmedialey iurn in lhs procedure, lhe ree immersed 23 thenpause for lhe developer times, 30 seconds.

Development pdntlngpaper ot
Thisseclon deals withdevelopmenl using
a tlay.

.l
I

2
X

/ ---

-\-

pr Whie holdlng edgeol the exposed nling lAs shoM in lhe piclure,do noi immerse lhe paperlighllywilh a pair of longs,quickly lhe prinlng paperinlo the developer parallel lhe developer place il inio the developer. to surface,bU till il nlo the deve oper balh.

lollowlhe developmenl, Alter completing ng iixlng, wash and sequence stopping, of

nol Thodeveloperwill bsut simplyby ficiently aoilared tuning lhet@|,andtheresull will be sporson ihe lilm,

lh By @movlng rcelfromlhe dev6lop.r,lhe between theroll6d dvlopscanreach spacs lh pvontlno oonnlingofspols. lilm, lhe

nlervals I the compielon I in 2 Pedorm operal shown piclure of ihe leftin 3o.second lhe on work. ol developmenl

paper I Make surethallhe photograplric of doesnol float!p lo the surface lhe

work Aftercomplelifg development on lhe prnting paper, cky pe.lorm sequence qu the fixing, washng. and ol stopping, sheels prifilng ot surelhatseparale I Make paper nol sticktogether thefxer if do I Do nol usethe samepar of iongsduring and lhalyourseddu.ng slopping lixing developmenl work. the Make surenolto codaminale develoP ef wth e lherslopbaihor I xer qud.

ro agilale the phologGphic paper, ho d il at a cornerwlh a pair oi longs and move r back and lorlh whlle I is submergedn lhe aeveloper.Repeatthis operalion in 15-second nterva unilldeve opmentworkis compe1ed. s

Trouble-shooling

Section4

P|obloms in linished pholos and lheir conction Poorcolorreproduction . . . . . . .. .. . . . . . . . . . .. .68-7'1 .. Blurremage id ..,,........72-73 Theimageis in focusbut not sharp.. . . . . . . . .. . . . 74-79 Objectsother than the specimen imageappeared on the tilm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .80-81 U n e v eb r i g h t n e s s n ........82-83

in Prcblems finishedphotosand theil conection

-Poor colorreproduction
1. Th beckgroundis coloEd (.dibhF)
Caul6

Cor'rctlon
.ljse flltels spclfled by the manu' lacturer.

Rama*s .Daylisht type: LBI!2N, 5,5mK, D settlng .Tungstn type: LBT,34mK T setting

Relr lo Daq Page48/ page 50

of The color temDeralu@ the lliuminatlon ls not malched to that of the fikn. .Lioht balanclngfllter specified by manulacturerls not usd.

Lamp \oltage ls too low (loo high).

.Raise (low60 lamp voltag

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

.BHS above&5V .BHT and BHTUat 5-6V .lf you have reachedthe desired color tempature, do not chanoe the voltage position. lf you want to chango lrieBity, u6 an ND fllt6r. .!!b recommendthat for iest photograplrythe voltage should be set lo the standad Dosition +1V whgnfi6t detednining lhE ootimum color temDeEturc.

Pag65(y page 54

l1v-rcllaoo l. too hloh{bluohu).

Blackenlngof lamp due to pre longed Uae

.Replace wlth a now lamp

2. The backgroundis colord (grdmagenta) Caus6 .A iilm dilferentfrom yourusual film lype was used. Conction .Since colorrendition varieswith different typesevenfromthe film samemaker, choosea lilm with for coloring suitable yourtype

Rmarks

Rfrto Deq
Page 51

.A iilm of the same typebutwith difterent emulsion number was used,

.li possible filmwiththe buy numbr laryo in same emulsion quantities.

.ln orderto maintain perforthe mancequalityof the film, slore it in a relfigoralor and remove it one ho0rpriorto us6,allowing it to reach room lemperature, .Even for film of the sametype in therewill be slightvariations on colof reproduction depending condllions, labofatory, development and stainingof the specimen.

Pag652

.l.Jsea colorompensating (CC) lilter

O ta

a a
t

t
t l

a rt
t

a
I
O -

oC

t
t

t
t l

ea B

ar I
t

'Colorompensatin0 tllters (CC) dealer, Buyfromyourpholo grn .Too much UseCcM {magnta) pink .Toomuch (gren) UseCCG usecc Forphotomicrography, filters For compensa0+10. lurlher and tionuseCC05 CClologether

Page53

+
ra

rt

cc05M-optlmumcompensallon

a a O
rt

:i. r.l

a
*

.a

cc10M-ov6r{ompensared

3. Inconeclcolor rendilion Caus .An excessively exposure long time hasbeenusedandthe characieristics reciprocity of law failureleadio incoftect color rendition. Conctlon .Set shutter sped 0.01{.05sec. at Adjustshutterspeed uniJormly as fal aspossible using ND{ilter. by an .When usinglongexposure limes (above sec)wilh models 0.5 PIV'1oAD PM'1oADS, the and set characterislics number the film of usedon ihe dial for reciprocity law failure, .When usingmodelsP[,]-1oADS/ PMJoAD the exposure set adiustmentdiallo: 0.8-0.25X bighl background for specimens to 1.2t4Xfordark and background specimens. .Carry out exposuadjustments Brlght backgrou ISO/ASA to 50 nd: 100 Dark background: ISO/ASA to 200 100 .Vary the ISO/ASA speedsetling. Remarks .With longexposure times,even wiih the exposurc time compensated, colorfendition the changes as a rcsullof film properties. Usea colorcompensaling filter speciiied the film manu' by Relerlo pag
Page 216

.Automaticexposure been has usedwithouladiustment.

.ll you change magnification Page4Z the of the objeclive, lightdistribu- page 43 the lion withinthe visualfield will atsocnange.

.Aclual and nominalsensitivity ol lhe iilrn differ

.Nominal speedmay diiJer lrom actuallight sensitivity much as as 1/3to 1/2aperture.

Page 42

lrom a negative 4. Poorcolorol a prinl enlargd

.The printing technician the in laboralory nol sureof lhe is subiect's correclcolor.

.Add a lransparency the same of specimen with correctcoloras a sample.

.Better coior is achieved when with shooting transparcncy a reversal colorfilm (butthe cost of pints goesup and quality

5. Poorcolorol Polaroid lilm Cau3e colortemperature. Coneclion .The film is a daylight type,but because the film characterisof tics colortemperature shoutdbe set lowerthan for 35mmfilm. Remarks .lf you st colortemperature b+ tween2rc00K 5000K and and use an Olympus filter,color FF contraslwill b enhanced. Rferto Page58

.Abnormalcolor characieristics.

.Term of validilyof the lllm has .ll roomtemperature abruptly is (raised), lowered sensitivity is alfectedand printcolorturns towardsblueted).

.lf the abnormalilies within are the adjusiment rangelor color temperature withinthe and operalional rangeol the colorcompensatlng lilter,determine ihe conditionsbytest photography .Use film whoseterm of validity has not yet expired. .For developmeni time and color reproduction, to the film refer instructions,

.Since Polaroid film is easily Page54 aftected by adversestorage page58 conditions far as colorcharacas protectit leristicsare concemed, irom heatandhumidity bystoring it in a refrigerator.

image Blurred
l. The ovrall locus ol the oicturc is blund Cau3 .Finder 6yepisce focusing or t6lescopeis nol prope y adjusted, Conlction .Adjust thediopter untilthedouble crosslinesare clearly visible.

Remarks
.Slnce most people not have do the samevisualacuityln both eyes,dlmlnewhicheyeyou alwaysuse for focusing.

page Pages 2e27

lt
ll
.Blur anddrittresulting from vibrations.
.Uss a vlbration.proof table.

dF
lm.o..fi6t.d by vlbEttons

.Uss an ND flltr and lncrease shutter speed (1/2sec to 1 sec). .Use a standfor the photographlc photoequipment separate to graphlc equipmenl microscope. and

Page12 paqe 48

2. Focuslngenor occurs whn you u96 a low.magnlcallonobiectiveol less than 4X

Ceus
.lf magni{ication low,tocal is depthat the lllm planebecomes erors shalloq easilycausing in focusing.

Coarcllon .Mouni a locuslngmagnifier to lhef Indereyepiece thefocusing or on lelescope, afterfocusing and fine focus the double crosslines, the specimen.

Remafts
.lncreasingthe magniticatlon tocusing erors, eliminate6

Reiar to pa96 Page27

3. The prlphryls unilomly blund Cau3 .An Achromattype objctive was uso. Coreclion R6ma|ks Reler to paq Pag14

ausea Plan Achromat type obiective

l,r,i';.,r', 1.,iil;.'

;;1..i.'i:.1 ;i:'-:: l,i;"li ;1

;';iiii,''''r,;ii'':.:r'::;ji liir'i'.1.t...'1,.:
:',:,1:,11i,;i;,,.,. .;.,.
;t::'. . - . _ :J : - - i : . . ! : - : r .

j;:. ::.:':itl:i: -r11.;'i : .ij, ':i

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

/:::).\

.'-

-::'

.):tt.

::

D Plan1oXNFK2-5X

.Objective and photo eyepiece were usgd in th6 wrong combination.

(longbanel)-NFK .LB series photo eyepiece ashort-barrel serles-FK photo eyep|ece

Page13

Theimage in focusbut notsharp is

1. Inadequat power resolving .Use ol a combination lowof magnification obiective highand magnification pholoeyepiece, .ln orderto obtainhigh resotuiion, usean objective with large numeicalaperture and a pholo eyepiece with low magnification. .ln orderto obtaina magni{ication

;i ioot i; tniirr. pti ", ,iJ-

Pase 38

-?

,1

zlox eitheran objective with a photoeyepiece 2.5Xor an of objective with a pholoeye 20X piece5X.To increase resolving power, combination obiecthe of tive40Xand photoeyepiece 2.5X i (Focaldepth, is pfeferabie. however, bcome will shallow)

.Use of the condenser with lhe apenure diaphragm iis fully opened.

.Stop downthe aperture diairis phragmto 600%

.Vary the amountby wh;chyou reduce theaperlure diaphragm iris according the magnificaiion to of the objective the contrasi and ol the specimen. .Do not reduce diameterof the lhe field kis diaphragm such an to extentthat it louchesthe frame reticlebecause aclualarea lhe photographed alwaysslightly is larger lhan the areawilhin the lramereticle. .Olympusobjectives biological lor specimens havebeendesigned in sucha way ihai optimumresolvingpoweris obtaaned when a coverglasswith a lhickness of 0.17mm used. is

.The field iris diaphragm fully was opened,

straylight,slop downihe field iris diaphram an to areaonly slightly largerthanlhe .Use a coverglasswith a thick. nessof 0.17mm.

.Use of thick coverglass.

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

'

.. '.t

Cause
.The specimen slain is too weak, resulting lack of conlrast. in (Photographingspecimen a that cannotbe stained)

Corction .Use a denserstain, .Use a contrastlilter (fof black-and.whiie pholography only). .ll the specimen inteF cannolbe slained, phasecontrast, use differential iercncecontasl or darklield createconirasloDticallv. to

Beler lo pag6

Page57

gee
G*,il
oiiiGnlial inlorfercnce conlrasl

_Effiil ST

Cause
Colorphotography .Use of a low convasifilm,

ConEctlon

Remarks

Refrto peg Page51

.l.Jsa high conlrastfllm.

tt))b,;nite-:i

:',

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

i:);'.':ii;i;?;":i
photography Black'and.whlte .Variations lhe soclralsnin sitivityof the film ifiect ihe Page57 .Normally, greenfilter is used, a but if you want to emphasize a spcificportionof the spclmn, use another contrastflller, .lf a filtercomplementing color the of a specimen used,it will is emphasize contrast. lhe

2. The image appearshazy

Caus
aTheconection collarof the obto iectiveis not adlusted the lhickness the covrglass, of ,i , . r, 'Ja. . 'rt .

ConEctlon .Adjust the conection collarwhile xamining specimen set the and it at a positionproviding cl6ar a image.

Rmarks
.On the LB objective correction a collaris mounted the S Plan on Apo 40X S Plan100X and dry D PlanApo 60X.

Rolor to pags
Page16

j .:-1 'l: i ,' '-r',!! ba . ' . r

. ' l r .

Cause .An objective normally usedwilh specimens was cover{lassed ussdon a specimen withoutcover glass(orvice vefsa),

Concllon

Bema*s LB objectives .NC S Plan40X .NC D PlanFL mX .NC S PlanApo 1mX oil .NC S Plan100X dry

Relgl lo pags

.Use a no{over objective. ' ji;t,

Page17

'\ )/ '-. J .'1\)/'U ' ,. 4 ) . . ^ \ J \ - )-

) \.'1t

-.J

''(t

-\

!i - - - , , ',--'(.- \ ' r '- v: - 'J .-.t .

.Finqenrints,an oil lilm or dirl particles the opticalsyslem in (objective front lens,photoeyepiece,prism,specimen, etc.) .Clean lhe oplicalsystem. .Alwayscoverlhe with microscope a dust coverwhen not in use. .lt you use dry 40Xor 60X objectives togetherwiih an oil. ths immersion objective, oilon ihe may soil the fronl specimen lensesol lhe dry objeciives. Pages 26- 32

!.-

..,t,
1: ,', .'

3. No sharpimag6is obtained wilh a 10OX olflmmerslon obiective Cau36 .No immersion was used, oil .Unsuitable was used, oil Conection .Use oil specified the manuby facturer, .Use specified sincethe types oil, and usedfor fluorescence nomal white light oftendiifer .Apply th oil aftryou hav removed bubbles the botlle. ln .lf roomtemperaiure eithertoo is highor ioo low,o. if the air is too humid, diffiactionindsx the changes, causingchanges the in imag, Usethe oil at a room temperatureof 22-25"Cand at a humidily about56%. of .lt is advisable usea specimen to with a thickness 2'3r. of .Removelhe eyepiece before examinalion lookthrough and the Remarks Rl6rto page Page18 Page18

.The oil contained bubbtes. air

Page18

.Usingoil in a roomwithunsuitable (too temperatures highor low)

Page12

.The specimen too thick. is

l,

r1S:.
'.o'*.

-j- p
!.'l

lllm 4. Entireroll ot black.and.whlle ls nol sharp

Cruse
.Possiblecausesare:type of film, emursrcn, overexposure, overdevelopment, improper handling or accidents duringdevelopment,

Conction .Check lhesepossibililies and

Rema*s

Relerlo oaQ6

Pages

B{- e,5

pdnl appars grainy 5.Th6linishgd

Caus
.A fllmwlthcoarse orain wasosed,

Coreclion .Use a finegrainfilm.

Rmark5 .Kodak Panatomic X .Kodak Technical 2415 Pan .Agfapan25 .lliord PanF

Referlo paq Page55

.A slandard was develooer used.

.lJse a linegraindevelopet.

.Kodak Microdol D-23 D-25 .Fuji MicrcFine

.Magnitication ratiowastoo high.

aL,se largsformat film. a

.4" x5" sheetfilm.

image Objects other thanthespecimen appeared thefilm on

'1.Shadow.llk lmag

Caus
.Optical palh selector the ot pholographlc attachmgnt vinc or cularlube was intenupled at

Conction .Engagethe opticalpathselector posllion, at its propr

Remarl(s

Feterto
Daqe

.The lield kis diaphragm was stopped downtoo much.

.Open the field kis diaphragm a the littlewiderthan photographed of areaot the lindereyepiece the focusing telescope,

Page23

.Tlny bits of film, dirt,etc. stuck to the p sm of lhe photographic equipment to the largefomat or

.Chck for, and remove, from dirt the prlsmot the photographic atlachment whilethe shutleris open(Iime setling). .Flemovethe largefomal relay lens and cleanit.

.Periodicchecksare recomof mended a largenumber if photographs taken, is

Pages 2931

Cau3e .Dirl In the opticalsystem

Corectlon alocate the dirt and remove it.

R6ma*s .Yo0 can locatethe dirt by moving and rotating eachcheckpojnt, altnatelylooklngthroughthe blnocular tube,the focusing telescope the photographic of attachmsnt the lllm plane and (by placlnga pieceof frosted glass in the camsra body.)

Rlerlo Page PageI Pages 30-31

Cause
.When movingthe film or when the unrolling backingpaperof causes the fllm, staticelectricity spafis.

Conclion .Do not rewind film too rapidly. lhe .Keep humidity zl5%minimum at in the roomwhereyou handle the tilm. aMakesurethat the camen back and the darkroomars lree of dust,

Rl6rlo page

3. Rellocllon windowor roomilluminatlon ol

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

Coneclion .Move lhe oplical path selector of the tdnocular tubio lhe Camera 1m7oposilionandcover of the locusingtelescope the photographic atlachmenl wilh a cap. aPut capson both eyeplces and of the locusingtelescope the photographic atlachment. .Set up the mlcroscope a in dlllerentlocation.

Referto Pag Page12

Uneven brightness
1. lJnyenareas occu. on one sld6 of th6 fram, in lhe cntr,and under the pertoratlonof th film

Cause
.Th microscope light sourceIs not prope y centered.

Conclion .Prope y adjustthe light source.

Rma*s

Rlerlo paqe

.The field iris dlaphragm off is axis.

.Clos th6 fleld his diaphragmso that it appearsin the visual field and adjust the conden6erto center the diaphragm. .Clean the opticalsystem.

Page21

.The optical syslem is contaminated by dirt.

Pages 29-31

.Dgvelopment problems(on blackpint) and.whlte

.Developthe film prcperly.

.8e aware lhal a change In l6mpeture occurs betweenthe center and the pedpheryof the tank as a resultof hgatconductlon by th6 slliank

Pages 64-65

2. Ma*s on the nogative

Caus
.Fixing time was too shortor exhausted fixer was usd.

ConEction .lncrea6etixingtime or use a new lixet

Roma*s

RaLr lo paEe

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