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AH3-NAF AH3-NSF AH3-ANF

Nomarski-type differential interference-contrast attachment for use with reflected

light fluorescence attachment

WARNING

This instruction manual has been prepared for the transmitted

light Nomarski differential interference-contrast attachment.

In order to gain optimum performance also read the assem­

bly manual and the instruction manual for observation and

photomicrography provided with the VANOX microscope

and the instruction manual provided with the reflected light

fluorescence attachment AH3-RFC.

OLYMPUSe

CONTENTS

1 OUTLINE

2 COMPONENTS

3 COMPATIBLE EQUIPMENT I OBJECTIVES 1

4 SPECIFICATIONS 2

5 NOMENCLATURE 2

6 ASSEMBLY INSTRUCTIONS 3

7 OBSERVATION 6

8 TROUBLESHOOTING GUIDE 9

1

2

3

OUTLINE - . . Th1s unit rs a polarr7,ng-type 2-beam shearmg d:fferent1al rnterference contrast attachment us,ng b~refnngent crystals as beam sp rtters

Used wr!h the reflected 1Jg11t fluorescence attachmeN, m1nute vanauons 1n light of unstarned or transparent soec1mens and smal changes rn optrcal densr:y graarents become detectable. Used dunng fluorescence observation. the attachment 's

especral y useful, srnce 1t makes 11 easy to observe ooth fluorescent and non fluorescent deta1l of the spec1men.

COMPONENTS The attachment cons1st of the follow1ng 3 modules.

OB side Nomarski slider AH3 NAr

CD srde Nomarskr sllder AH3 NSF J~

Analyzer slider AH3-ANF

COMPATIBLE EQUIPMENT/OBJECTIVE • Compat ble uMslattachments Research pho:om1crograph1c m1croscope systems AHBS3, AHBT3

RerJected ltght fluorescence attachment AH3-RFC

• Compallbl(; OOJecuves DPianAoo Senes ldry ob1eCt1ves onlyl * When using the SPian Series objectives, the Nomarski slider provided with the transmitted light Nomarski differen­

tial interference-contrast attachment AH2-NIC should be used instead of the CD side Nomarski slider.

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4 SPECIFICATIONS I . . . ' , , • . .. . r , • ,. · ~ • ~ . . •

Module Description

OB s1de Nomarski si der AH3-NAF • Nomarsk1 onsm movable by means of

•otat on knob i-

CD s1de Nomarsk1 si der AH3-NSF • 6 POSitiOns poss1ble Nomarsk1 pnsms. 4 pes

110x, 20x, 40x, 100x),

polanzer. empty open1ng

A~alyzer slider AH3-ANF • Analyzer. depolarizer bUilt-In

• 360° 10tatable by means of d1al

5 I OB side Nomarski slider I

I CD side Nomarski slider I Prism movement knob

Polarizer I Analyzer slider I

For 20x prism

For 1 Ox prism

For 1 OOx prism

Analyzer rotation dial

For 40x prism

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6 .ASSEMBLY. INSTRUCTIONS .. I • . ~.~, ~ , ,_t .. ·~t~ ... ~c .... , .. ~1 .. ,.·\ ~ .... _<:l'.' ··

6-1 Assembly Outline

* Check all connecting surfaces for cleanliness and avoid scratching the surfaces during assembly. * For details on the microscope main-units, refer to the instruction manuals provided with the AHBS3 or AHBT3, and

AH3-RFC.

CD side Nomarsk1 slider

rn Inserted from the left hand side • 2 Inserted from the front

Fig. 1

OB side Nomarski slider

1 3 • Inserted from the right hand side

* This attachment is designed for use with DPianApo objectives. If any other type of objective is attached, replace 1t w1th a non-immersion objective of the DPianApo Senes.

* When using objectives in the Splan Series, the Nomarski slider provided with the transmitted light Nomarski-type differential interference-contrast attachment AH2-NIC should be used instead of the CD side Nomarski slider.

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Fig. 2

Fig. 4

Fig. 5

D Attaching the CD side Nomarski Slider

I 1 I Remove the covers attached to both s1des of the stage holder !Ftg 21 * Using a coin, etc., makes it easier to turn the fastening screws.

!21 Rotate the condenser turret until "20-10" or "100-40" IS d1sp1ayed

* If the condenser turret is at the " 4 - 1" position, the slider cannot be inserted.

131 Insert the CD side Nomarsk1 sllder@ from the slot on the leh hand s1de of the stage holder !Ftg 31

* Malfunction will occur if the slider is inserted from the right hand side.

fJ Inserting the Analyzer Slider

( 1 l Remove the dum my Q) from 1 he f ron l of the fluorescence nosep1ece. !Ftg. 4)

121 A tgn the ana yzer ~ wtth the slot ar"'d msert n !Fig. 51 * At the first click-stop the analyzer is not in the light path. At the

next click-stop the analyzer is engaged.

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•

\

I \

Fig. 6

Fig. 7

EJ Attaching the OB Side Nomarski Slider

( 1 l Remove the dummy(.]) from the nght stde of the fluorescence noseptece. (Fig. 6)

(2) Align the OB stde Nomarsk' shder (2) wtth the slot and 1nsen 1t (Ftg 7) * At the first click-stop the slider is not in the light path . At the next

click-stop the slider is engaged.

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7 OBSERVATION .

7-1 Preparation for Observation

L

* Preliminarily adjust the VANOX microscope unit and the reflected light fluorescence attachment. For adjustment of the VANOX m1croscope refer to the provided "ASSEMBLY MANUAL" and the " OBSERVATION AND PHOTO­MICROGRAPHY MANUAL". See the instruction manual provided with the AH3-RFC for details on adjustment of the fluorescence attachment.

F1g. 8

Fig. 9

( 1 l Slide the analyzer CD tnward to the last click-stop position (Fig. 81 (21 Sltde the CD stde Nomarsk1 sl1der ® all the way to the right until "P"

appears on the nght s1de of the stage holder. (Fig 91 131 Remove the pnsm from the optical path by push1ng the OB side Nomarsk•

sltder @ to the ftrst click stop positton (Fig 8) (4) Use the cube change switch @ to select and engage the bnght-field

(BFl cube fF,g. 8)

(51 Push the OBJECTIVE mag. button on the VANOX microscope to engage the 1 Ox objective

(6) Whtle observing through the eyepieces, turn the rotatton d1al @) of the analyzer \!) Stop at the postt ton where the f1eld of vtew tS darkest. (Ftg. 81

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•

I

7-2 Nomarski Differential Interference-Contrast Observation

!How to effcct•ve1y uuhze t!"'le d fferenttal interference-contrast attachment]

1 Rotate the AN pnsm to search for the pos•uon offenng opumum sensn•vtty

2 Fteld ns dtaphragm should c~rcumscnoe the v1sual fteld and the apenure 1ns dtaphragm should be stopped

down to leave only 70-80% of the ob1ecuves effectiVe aperture open to prevent reduced contrast due to

flares. I fhts sett ng 1s CJutornattcally performed on the AHBS3 l 3 Dunng photomtcrograpny, photographs wtth 1ttle unevenness can be obtatned by ustng a 5X photo eyepiece

-1 W hen taktng 8/W photographs, photos with improved contr<Js t can be obtatnod by using the IF550 fl ter

5 Detoctton scns1tivny of differenttal Interference-contras t is htgh, so parttcular anentton should be g1ven to d•rt on the spectmen surface.

. . '" .. ~ ........................................................................ - ............ .... - ................................ -- ---- .... - .... ---.- .... - ........................................................................................................................... J

Fig. 10

Fig . 11

Fig. 12

I 1 l Pull out all filters from the light path (Ftg 10)

12! Ergage the bnght-fleld cube and objective to be used

* When the field of view is dark, set the low voltage selector switch to MAX.

13) Pos lion the CD Stde Nomarskt slider accordtng to the magntftcatton of the

ob]ecttve to be used. (Ftg I 1 l • The numbers "10", "20", "<:10" or "0" on the sltder will appear altgned

With the left s1de of the stCJge holder, " P' " and " 100 '" w ill appear on the right side

14! Slide m tne OB s1de Nomarsk• sl1oer CD to the last cl ck-stop to bnng the onsm 1nto the I ght path (Ftg 121

151 Changing the background calor

Rotate the pr sm movement knob ( 2) of the OB side Nomarski slider to

change the Interference calor of the background continually from 0-530 nm (black to sons1t1vc colors) (Fig 1 ?I

Depend ng on the spec rnen, choose the backgrourd calor w h1ch results 1n opt1mum contras t tor tne observauon.

• If the background calor IS dark, dark·fleld-like obseNatlon is poss1ble • If the background calor IS gray, observation w1th optimum sens1 t1v ty

becomes possible (Gray contras t)

• If the background color is purplish red, observatton uti lizing calor change

.s possible CJ iso 1n the case of mtnute phase difference !Sensitive color)

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Ftg. 13

161 Improving contrast • When observ ng uSt'1g gray contrast. contrast may be tmproved by ustng

a green filter

• To obtatn better contrast. the aperture tns dtaphragm may be stopped down to leave only 60 70% of the ob)BCttve rear aperture open However, resolvtng power decreases when the aperture 1ns dtaphragm ts stopped down

• Wnen rt ts destrable to tntenstfy contrast tn the center portton, stopp ng down the field tns dtaphragm wtll reduce stray light from the penphery of the vtsual field, resulting tn improved contrast.

(7) Rotating the specimen

The detectton sensitiVIty of the dttferenttal tnterference contrast IS affected by the onentatton of the spectmon When the onentat1on of the spec,men 1n the field of v1ew IS as shown 1n F1g 13, oot mum sens1t1v1ty ts obta1nerJ Rotate the stage to arrAnge the d roct1on to be observed as shown 1n F1g '3 * If the stage is not centered, the observed detail may disappear

from the v isual field when the stage is rotated. In this case, center the stage by referring to the " OBSERVATION AND PHOTO­MICROGRAPHY" and " ASSEMBLY" instruction manuals of the VANOX microscope.

7-3 When other microscopic examinations have to be performed

I I l Pull out the analyzer sllcler and OB s1de Nomarsk1 slide' to thetr ftrst click-stops to remove them from the light path 121 Pull out the CD s1de Nomarskt sltocr to the left until "0" appeors at the left hand s1de of the stage holder

7-4 Used together with reflected light fluorescence attachment

* Refer to the instruction manual provided with the AH3-RFC for details on the combined use of the reflected light fluorescence attachment and the Nomarski-type differential interference-contrast attachment.

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8 . T.ROUBLESHOOGING GUIDE If opt1mum performance of th1s attachment cannot be obta1ned, please consult the table below for troubleshooting

Problem Cause Remedy -

a) Interference calor does not appear OB side Nomarsk1 slider 1nserted into Insert 1n to correct slot wrong slot

CD s'de Nomarsk1 shoer nserted 1nto Insert 1nto correct slot wrong slot

-bl lntenor contrast Improper sett1ngs of polanzer and AdJuSt se111ngs.

analyzer

c) lntericrence calor appears but color Obrect1ve magn1f1cauon and magn1fica- Match correctly. unevenness is consp1cuous lion of the CD s1de Nomarsk1 slider do

not match.

Objective used is not DPianApo Senes. Replace w1th DPianApo Senes' obJeCtive ~

Condenser pos1t1on IS 1mproper Center the condenser properly. f-

dl Nomarsk1 effect IS tnfenor Onemauon of spec men and sheanng Rotate the stage to f1nd a good pos,uon

d1recuon are not 1n agreement for the obseNat•on

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OLYMPUS OLYMPUS OPTICAL CO., LTD.

San-Ei Building, 22-2, Nrshi ShrnJuku 1-chome, ShrnJuku-ku, Tokyo, Japan

OLYMPUS OPTICAL CO., <EUROPA> GMBH. Wendenstrasse 14-16, Postfach 104908, 2000 Hamburg 1, Germany

OLYMPUS CORPORATION 4 Nevada Drive, Lake Success, N.Y. 11042-1179, U.S.A.

OLYMPUS OPTICAL CO. <U .K .) LTD. 2-8 Honduras Street, London EC1 YOTX, United Kingdom

rtw dcsiQP o f trle product is uqder constant rl'vi<'W <.~rnJ whils t Pwry ellort is r•wde to kcco this manual up to date. the ogtn is reY•rvu<.J to L'Mr·qe spe<.:'frL<l· trons and equipmPnt at any r·me without prior notrrr'

Pnntea '" Japan 91 12 M 0 I