Nikon Eclipse Ti Series User manual

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GGLIPSG 'Ti Series
Differential I nterference
Contrast Attachment
Instructions

lntroduction
Thank you lor purchasing a Nikon product.
This in;truction manual is written for users of Nikon Differential Interference Contrast Attachment for the
Inverted Microscope "ECLIPSE Ti Series."
To ensure correct usage, read this manual carefully before operating the product
. No part of this manual may be reproduced or transmitted in any lorm without prior written permission
lrom Nikon.
. The contents of this manual are subject to change without notice.
. Although every effort has been made to ensure the accuracy of lhis manual, errors or inconsislencies
may re-main. liyou note any points that are unclear or incorrect, please contact your nearesÎ Nikon
reDresentative,
. Some of the equipment described in this manual may not be included in the set you have purchased.
. lf you intend to use any olher equipmenî with this product, read the manual tor that equipment too.
. lf the equipment is used in a manner noî specified by the manufacturer, the protection provided by the
equipment may be impaired.
Notes on Handling the SYstem
1. Handle the product gentlY
This product is a precision optical insîrument. Handle the product gently, avoiding any physical shocks'
In particular, the optical system used on DIC method must be kept strain-free; handle the objective and
condenser carefully so that they are not deformed.
2. Dirty lenses
Do not lel dust, fingerprints, etc. get on the lenses. Dirt on the lenses, mirrors, etc. will adversely affect
the view of the image. tf any of the lenses gets dirty, clean it as described in Chapter 6, ' Care and
Maintenance."
* 3. Installation location
In order to avoid degraded performance and to prevent malfunctions, take the following rcquirements
into consideration when selecting a location to inslall the product:
. lnstall the product in a localion with little vibration.
. Avoid installing the product in a location exposed to direct sunlight.
. Avoid installing lhe product in a dusty location.
. Avoid installing the producî in a location subject to high temperatures (40"C or higheo or high humidity
(60% or higher). (Such conditions could allow mold or condensation to form on the lenses and filters.)
4. Specimens
Use slide glasses and cover glasses that are free of dirt, scratches, and distortion when preparing
soecimens.

5. About this DIC attachment
This Dlc attachment employs the "senarmont system,' in which the contrast of the Dlc image is varied
by the direction of the Polarizer.
The Dlc prisms ar" ro o" intt"tt"d nearthe obiective and the condens€r lens. This attachment requires
the correct combination of the objective, condénser lens and the DIC prisms. Read the following notes to
. choose the correct combination.
For condenser cassettes (Dlc prism fof condenser) of lhe '@r, "oM", 'oH" or "ooSS" marking' use
the following combinations.
DIC prisms for obiective :
Dlc prisms for obiective and objectives work in specific pairs. use the Dlc prism for objective Îhal
has the same name as lhe obiective.
Condenser casaettes (DlC prisms for condenser) :
The condenser cassetle to be used (the Dlc prism for condenser) is determined by the NA
(numerical aperture) of the obiective and the type of the condenser lens' (The NA of each
obiective is indicatéd on the decorative ring of the obiective')
For LwD condenser lgns
NA < 0.5 @ L (blac* lett€rs on whlle base)
0.5-NA<1.0 @ M (blad( letbrs on whlia base)
1.0 s NA o. H (Uack letbrs m whits bas€)
For dry type high NA condsnser lens
0.5<NA<1.0 @ M (Yvhite lstlars on gre€n base)
1.0 s NA 6H (whlte letters on gre€n bas€)
For ELWD condonser lcns
NA < 0.5 -L (whlte letters on red base)
For oit type high NA conden3er lena
1.0 s NA oH (blac* lettars on lrellon baso)
1.0 = NA - SS (black letters on ysllovi, base)
Zmark:
rne otc prism tor objective having the z mark (Dlc pfism for Plan Fluor 10x), and the Dlc
prism for condenser Àaving the I mark (LWD condenser oL) should be used in pairs.
(2) Combination lor Nl / N2 / NR tYPe
r* cono"nser cassettes lDlC prism lor condenser) ol the "N1', "N2', or "NR' marking, see Table A
onthe next page and select the oblective, DIC prism for obiective, and DIC prism for condenser
(condenser casgette) resP€ctively.
òtandard combinalions, high conirast combinations, and high resolution combinations are listed. Use
the appropriate combination.
For EiWó objeAives, only one type of condenser lens can be used. See also Table A on the next
page lor the ;ombinaiion ót eLWD objective, DIC prism for obiective, and DIC prism for condenser
(condenser cassette).
2

Notes on Handling the System
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3

GontentS
-
lntroduction I
1
Notes on Handling the System
.5
Names of Parts 7
Microscopy
Princ3ples of DIC MicroscoPy
1.
2.
3.
4.
5.
6.
10
12
17
18
AssemblY
TroublÉhooting Table
Care and llaintènance
4

Names of Parts
lf the system has not yet been assembled, refer to Chapter 4, ,,Assembly," first.
For details on the assembly and handling of the microscope, power supply, etc., refer to their respective
instruction manuals.
(The Nikon Inverted Microscope ECLIPSE Ti-E with the DIC anachment mounted is shown below. Some
components may not be included in the set that you purchased.)
T-P2 DIC oolariz€r
(Slide to lhe lelt to bing ths
polarizer inro th€ oplical
path.)
Systom condeng€r
T|-NDG sextupb DIC nosepisca
or
T|-ND6-E sèxtuplo motorizsd
DIC nosópisce
of
Tl- D6.PFS perfa€t focus with
motorlzed nosépl€ce
(Athch a DIC obj€ctive and a DtC
prism for objective h€re.)
(Attach the DIC condenser
cassotte hsre.)
ffh€ro qrq lwo iypss of turrets:
a manually oporat€d ong and
the TI-CT-E motoriz€d
cond6nser turret.)
DIC prilm lor
obirctlv€
Th€ prcduct can b€ us6d with any ot the following
lhros eDl-fr filter turrets:
. TI-FLC ept.ll fitter !.rret
. TI-FLC-E motoriz€d epi-f filter tunet
. TI-FLC-SFIO motorized eplf, fitter turet
Figure l-1
Push in ìo bring the anatyz€r
into optlcal path.
Vibration
direction lever
clamp scrow
(bottom)
5
I o,.oo,"*r"
T-N2 anaayz€f
or
Epl-ll tllter turrot
Ol-A Analfzer Block is attached in the
turret.)

Chapter i Names of parts
T-P2 DIC polarizer and sysfem condenser
Rotation
Dial scalo (5. intervats
Whit€ ind€x nark
When using a high NA condenser tens
Condensor ap€dure
lsvsr
Sllder lever
Dul-proof plat€
Flt"",J3î?H"là""'mF
rufrfi{ffi$F$""d.i{i
Polarizer
s€cton
Polarizer rotaiion lev€r
Condensor tJret
Condgnsor screw
lever
Ext€nsion ùJbe
Condonsgr slider
Condgns€r
Figure l{
6
Figure I -2
Slider damp screvi,
High NA condonser tens

Microscopy
Focus and cenler the condenser as described in the manual provided with the microscope, and if lhe
Tl-ND6-E sextuple motorized DIC nosepiece, Tl-CT-E motorized condenser lurret or TI-FLC-E
motorized epi-fl filter turret (or TI-FLC-E/HQ motorized epFfl filter turret) is being used, also refer to the
instruction manual supplied with the motorized units forTi series.
. Inserî the "A" (empty) cassette into the optical path. (ll using the condenser slider, remove the
condenser cassette, or move the slider so that there is nothing in the optical path.)
. Remove the polarizer, the analyzer, and the objective DIC prism from the optical path.
. When using a high NA condenser lens:
In order to focus the field ap€rture diaphragm image on the specimen surface, and in order to
avoid striking lhe specimen with the condenser lens. move the condenser lens slightly closer than
the subiect distance for each condenser lens, and then bring the specimen into focus while raising
the condenser lens. When using a oiFimmersion type condenser, also check page 9.
DIC microscopy requires that the specimen containers, slide glasses, cover glasses, etc., be largely
free of optical distortion. Therefore, a container that will place plastic in the optical path is not suitable
for DIC microscopy.
In addition, if a direction adiustment is made in the optical system beforehand, it will be possible to
observe the DIC image with the besî contrast. (See page 15.)
Move the DIC l ox obiective into the optical path and focus on the specimen. (Stopping down the
aperture diaphragm slighîly will improve îhe contrast of the specimen, making it easier to bring il into
focus.)
Turn the condenser turret and bring the adequate condenser cassene for the 10x obiective into the
optical path. (See page 3, Table A.)
.N
til
1
2
Rotialion clamD scrgw
Figure 2-l
tr Loosen the rotation clamp screw of the polarizer and then
r, gently rotate the polarizer to change the image contrast.
. The highest contrast is obtained by adjusting the viewfield
background to sensitive gray and then bringing the GIF filter
inlo the optical path.
. Because contrasl is obtained in the shearing direction
(see Figure 2-2), adjust the orientation of the specimen until
conlrast is obtained in the desired direction.
Viblalion direction
ol analyzsr
ll 9o', Shearing direclion
llXou"
Vibration directton
ol polarizer
Figure 2-2
7
Table 2-l
1.gmm (including slid€ glass with a thickness of f .2mm)

Chapter 2 Microscopy
6Changing the magnification
Ex. 1: DIC LWD 60x NA 0.7 and @M
1 . Rotate the nosepiece to bring the DIC LWD 6Ox obiective into the optical path.
2. Because the NA of the Dtc LwD 6ox objective is 0.7, rotate the turet to bring the condenser
cassette labeted'DlC 0.5 - 1.0', into the optical path.
3. Adjust the contEst by rotating the polarizer.
When with ification
Focus on the specimen using a low-magnification ob,ective (1ox). with the specimen in focus, contirm that
(1) the field aperture diaphragm image is corecfly focused on th€ specimen sudace, and
(z) rne same tmage is corecfly centered in the viewfield.
lf eilher of these is not true, re-adiust. Once both are true, switch to the high-magnification obiective.
when observing a specimen with a high-rnagnif ication obiective, such as 6ox or 1oox, first correcfly focus on
the specimen, and then re-ad.iust the field aperture diaphiagm image on rhe speciren .urt""à. -Éy oorng
this, you can easiry re-focus on the specimen by focusing o-n the riÀrd aperture diapt ,"gm ir"é". '
Color contrast microscopy
Flgurc 2-3
1 . Rotate the nosepiece to bring the plan Apo 2Ox oblective into the optical path.
2. For. the standard and good balance view, bdng the "N2 Dry, condenser cassette into the optical
path' For the high contrast view, bring the "Nr Dry" condenser cassette into the optical pat'h. lsee
Table A on page O.)
3. Adjust the contrast by rotating the polarizer.
Remove the dust-proot plate from the DIC polarizer and insert
the lambda plate.
Rotate the polarizer until the background color changes to lhe
sensitive red-violet, a specimen with variations in itslef ractive
index or thickness will show interferenc€ colors corresponding
to the slope of the ì/ariations, producing the highest color
contrast.
To observe the imag€ with the correct color temperature, slide
the NCB filter into the optical path, and adiust the brightness
adiestment dial to match the lamp rating. Use the NDfitter to
adjust the brightness of the image.
Dtc
8

Chapter 2 Microscopy .f
ffi
NA 0.85, subiect distance: smm
Because the lens tip is cut at a 45' angle to the specimen, a wider work space can be attained.
When using the manipulator, rotate the lens rolaîion ring so that the cut surface of the lens faces the proper
oosition.
Use with Nikon-specified oil filling the space between the slide glass or cover glass and the lens'
Soread an amount of immersion oil approximately the size of the condenser lens tip on the side of the slide
gàss or cover glass that faces the condenser. Be careful nol to create air bubbles in the oil' The oil should
be about l mm thick.
Also apply a small amount of immersion oil on the tip of the oil-immersion objective.
After focusing the field aperture diaphragm image on the specimen surface, tighten the condenser refocusing
damo to mark the lower limiî for tho condenser. This prevents the tip of the condenser from hitting the
sDecimen and also fixes the field aperture diaphragm image.
Bemove lhe DIC polarizer, analyzer, and obiective DIC prism from the optical path before starting
microscopy. Because doing so will increase the brightness of the image, use ND filters or the brightness
adjestment dial to adjust the brightness.
NA 1.4, subiectdistan@
I

Principles of DIC Microscopy
since the human eye, cameras, and film caprure images by recording differences in light intensity and cotor,
they do not register colorless, transparent cells or baCreria. Although dyes can be used to make these
transparent subjects visible' the dye itself kills them. The differentiàt inteterence "onira.i iòio*i,ni"ro""op"
was deveroped in order to permit the observation of transparent subiects whire they are .tii "ìiul.
The basic structure of the Drc microscope is the same as that of an ordinary microscope, except that it
includes polarizing plates and wollasîon prisms'. inese ffiat erements change a transparent subject into
different levers of right inlensity. In other words, wnen yoLr ioot inrough a Drc microscope, you see even
]!Tqlt:tt tYb.,."cts as jmages consisting of different rivets of tighr inìtensity. 1-ruomaÀiibíi"iJ-"r" "rro
used as a variation of Wollaston prisms.)
The principles of a DIC microscope are discussed below
Light refracts as it passes through a subject, even if it is a colorless, transparent subiect. lf the refractive
index of a subject is ditferent Írom that of.its surroundings, light thar passes through the subject will reach a
ceiain destination point faster (or sloweo than light thaipaéés through the sunounding material. In short,
when lighî passes through a subject, it undergoei a pnaée cnanje lwitn me pnase eitÀJ, jJffi an"aa o,
being delayed).
A subjecl whose only effect on light is a phase change is
called a "phase subject." See Figure g-l on the rightl A
wavefront of light that is originalty in the same phise.
indicated by a single straight line, changes when it passes
through a phase subiect, so that onty th; light lhat passed
through the subject traveled faster than thelight that did not.
This phase change happens when light passés through a
mtcrosoope specimen, with the light passing through the
specimen traveling either faster or slower than the-
surrounding light.
Now look at Figure 3-2. This diagram illustrates the
principles.of a DIC microscope. In the diagram, the light
source is located on the left, so the light travels from Éft to
right. A polarizing plate and a Wollasion prism are each
placed on the condenser side and the obiective side.
When the light from the light source passes through
pol3îiilS plate 1 (th€ potarizeo, il wiil be potaize-cr (changed
to light that vibrates in one direction onty). When thià
polaized light passes through Wo aston prism f , it is sptit
into two rays that have mutually perpendicular planes oi
polarization (direction of vibration); the two rays travel at a
slighl distance from each other.
Polarlzing plate 1
Figure 3-2 Principles of a DIC microscope
since wollaston prism 1 is on the front focal plane of the condenser, the two rays pass through the specimen
in pafallel, at a slight distance from each other. The distance between the rays is called the ,shear." since
the shear is set berow the resorving power of the ob,ectivé, tne specimen does not appear as a doubre
rmage.
lftel,tlte lwo rays pass through the specimen, they are collected by the obiecîive onto the back focat ptane of
the obiective. wollaston prism 2 is placed here toiecombine tnà two rays 'uacr ino on".- Àorràu"r, the rays
do not intefere w*h each other since they have p€rpendicutariLnes ot poraization.
l9111zjno plate 2 (the analyzer) is placed so that th€ direction of potarization is perpendicutar to that of
polarizing plate I , taking out opposite phase planes in tne tight. where the speòiméÀ oio noì"i"ng" tn"
phas.e of the.light,.the two light rays intelere with """h otn"i, ,""r.ening them and m"iinf tno.à "r"". m rn"
vi€xffi€ld dark' where phase obiects-in the specimen cnangóo the phase of the righî, the two right rays oo
not interfere with each other, so those areas in the viewfield'appeai urignt. rnis oiieiencein urightness is
how the DIC microscope makes phase objects visible.
WavofÌont aft€r passlng
through spocimen
\
I
(
Wavgfroflt SD€cimen
>l
nl
Yal
lt
l subjecl
Pfrase
Figure 3-l Phase changas
10

Chapter 3 Principles of DIC Microscopy
of Dlc Mand Phase Contrast
,NF
ililil1
DIC microscopy and phase contrast microscopy differ in terms of their conÎrast and image characleristics.
Use lhe following table as a reference in order lo select the melhod that is best suited to the specimen'
Table &1
Ditf6r6nces in the oDtical thickness of line
structures determines ths brightness
conlrast.
The gradient of the optical thic*ness
determines ths color or brightness contrast.
Produces a threo-dimensional imag€.
Adjusted by changing objectives.
(Contrast varies according lo thé type of
phase plala and sp€cific absorption of non-
diffracied light.)
Adiusred by rotaling the polarizer.
Contrast adiustment
Fine obigcts are easily detecled.
Produces halo eflect.
Dstection s€nsitivity has no directionality.
Detection sensitivity is high but has
directionality.
Contrast is not alfected by subject size or
phas€ datè width.
Evsn specimens with a relativ€ly larga
Dhase differanca can be obsorvsd.
No halo eltect.
lmage charactèrislics
and detection sensitivity
Suitable for subiects with line struclures.
Phase difference:
DL lambda/4 or less
OM lambda/8 or less
BM lambda or less
Best results I thickness is nol groater than
10 r.
Suitable for structures of any size, whethef
fine or large, ewn dyed specimens.
Accommodatgs phase differences of even
several wa\relongths. Best resulls if optical
thidc|éss gradienl is nol grsalor than lwo
ìl,avelengths.
Permits observation even for thickn€sses
up to 0.5 mm. (tissue slic€s)
Suitable sDecimens and
phase ditference
toleraìnce
Shallonrer than for DIC microscopy.
2 to 3 timès ths phase ditferencs (at high
magnification)
C€nler the annular diaphragm and the
phase plate accurately.
Match lhe dirsction in which lou want to
add contrast with th6 shearing direction.
(See page 7)
Use distorlion-lre€ obiectives.
Us6 Ko€hler illumination for b€st results,
and make surg that the light source image
is enlarged adequately on ths annular
diaphragm plane.
Use a culture dish, Pelridish, €tc., wilh
glass surlaces that are fr€e of irrggularities,
smooth and parallel.
Protect thé surtacos of cover glasses, slide glassss, condensers, and obiaclives from
boing dirtied by water, oil, lingerprints, etc. Do not us€ l€ns-shaped hole glass, etc.
Approximately as bright as DIC microscopy
under white light illumination; slightly
darksr undsr monochromatic illumination.
Slightly brighter than phase contrasl micro-
scopy (under monochromatic illumination).
11

Assembly
Before assembly, be sure to read all warnings and precautions in the instruction manual for the microscope.
Install the sextuple Dlc nosepiece, perfect focus with motorized nosepiece, analyzer, Dlc polarizer, and
condenser in your microscope by pertorming steps 1 through 8 below. lf you reciuire'a very frecise vibration
direction adjustmenl, perform step 9 as well.
For delails on assembling the microscope, installing the system condenser, perfect focus with notorized
nosepaece etc', refer to the instruction manual provided with the microscope, and if the .T|-ND6-E sexuple
motorized Dlc nosepiece", or'Tl-cr-E motorized condenser turrer are bèing used, also refer to ihe
instruction manual supplied with the motorized units lor Ti series.
the DIC focus with motorized
Place the s€xtuple Dlc nosepiece on the square groove of the microscope's nosepiece mount, sliding the
nosepiece toward the rear of the.microscope as far as it will go. Tighten the bolts with a neiajonat wrencn
to secure lhe nosepiece. Fhis step is easier to perform if yo-u remóve the stage first.)
the DIC
Lower the focusing mechanism' screw the ob.iectives into the revolving nosepiece so that the obiectives are
::ojfl^Îj]l"t"uting powerwhen the revolving nosepiece is rotated initre ctòcrwise oirection 1*'nen uieweo
from above).
Dtc
the
Remov€ the dummy slider locateddirectly below the Dlc objective. Replace the dummy slider with a Dlc
prism that corresponds with the Drc obje;tive. lttote that ttró type of Drb prism useo oepenos onìne
. wnen ustng the T-42 analyzer
Remove the dust-proof slider on the microscope focusing mechanism and insert the analyzer. The
analyzer enters the optical path at the second clickstop iosition. pulling the slider ori to ínì rir.t
clickstop position removes the analyzer from the opticat'pattr ano places the empty hole in the opticat
oath.
Typically, the analyzer is installed so that the vibration direction adjustment lever is on the right, but it
can also be installed with thè vibration direction adiustment lever on the left, if you prefer.
. When u6íng the epi-fl fllter tunet attachod wlth the analyzer block
Insert the epi-fl filter turret along the attachment groove froó the right of the microscope. Remove the
filter block port cover and secuie the epi-fl filter túrret with screws on the right and left with the
hexagonal wrench.
Insert theEnalyzer block along the guide groove, and then attach the filter block port cover again.
objective.)
Figure 4-1
1. lf the system condenser is attached to the condenser
holder, remove it. lt is not possible to instalt the DIC
polariz6r while the condenser is attached.
2. Mount the DIC polarizer on the condenser mount of the
microscope, aligning the DIC polarizer positioning pin with
the groove on the condenser mount. Turn the fixing ring to
fix the DIC polarizer in place.
When using the T-A2 analyzer
12

.t
Figure 4-2
Mount rolation
Figurc &3
Condenser Cassettes
Tungt
Hexagonal soc*ot hoad sctews
Flgurc 4-4
Dust proot
plate
otc.o.d"n"", B,:ijÎtr;
séfew Ìroles
Condenser aporture
diaphragm l€ver
Ph @nd€nser
cassotte
Flgure 4-5
3. Loosen the rotation clamp screw, hold the polarizer rotation
lever, and turn the rotating portion so that the white index mark
is aligned with the center of the dial scale. (At this point'
conf'r-rm that the steel ball on the tip of the clamp screw is in the
groove on the rotating section.) Tighten lhe rotation clamp
screw to fix the rotating section in place.
4. Use a hexagonal screwdriver to loosen the condenser mount
rotation clamp screw on the left side of Îhe condenser holder'
Tum the entire polarizer by hand so that the white index mark
lhat was fixed in place in step 3. is lined up with the \ryhite index
mark on the condenser holder. Tighten the mount rotation
clamp screw so that the polarizer is fixed in place.
The polarizer can now be moved in and out of the optical path
by moving lhe slider.
5.
Slide the condenser cassettes into the condenser turet or the condenser slider and then fix them in place
with two hexagonal socket head screws. Note that the condenser cassettes thaÎ can be used are
deîermined by the condenser lens being used. (See page 2, 3.)
. When uslng the condens€r turret
Slide the condenser cassettes inlo the lurret so lhat their NA or
Ph code numbers increase as the turret is rotated in the
clockwise direction (whon viewed from above). Fix the
condenser cassettes in place with two hexagonal socket head
screws.
When uslng the condenser slidar
Set the DIC condenser cassette in the opening on the side with
the condenser aperture diaphragm lever' (Sinc€ the inner
prism must be used in a certain direction, setting the
condenser cassette on the opposite side makes DIC
microscopy impossible.) Instead of a hexagonal socket head
screw, screw lhe slider lever into the left side of the condenser
cassette to lix it in Place.
Notes on dust-proof Plale:
The dust-proof plate is fixed in place wiÎh two screws. lf the
screw holes on the opposite side are used, the dust-proof plale
can be installed in the reverse position' This is useful if you
need more work space in front of the microscope.
13

1. Screw the condenser lens into the condensef turret or the condenser srider. when using a high NA
::id.:::],lTf T!!|t rhe extension rube on rhe round dovetal mounr on rhe top of rhe rurrer or sticrer
the condenser
and then fix it in place with a hexagonal socket head screw.
Condensgr lens
Condonssr
FiguE &7
Figurc 4-o
Figure 4{
2, Install the completed system condenser on the
mtcroscope.
Loosen the condenser clamp screw (a hexagonal socket
neao screw) tocaled deep in the right_hand hole on the
condenser holder.
Slide in the system condenser along the dovetail groove,
and then fix it in ptace by tightening the condensei ctamp
screw.
When using the condenser turret, fix the system condenser
in place so that the indication on the condénser,".."tt" -
above the condenser lens faces the front. When using the
condenser slider, fix the system condenser in place sòthat
the indication on lhe condenser cassette on the side with
the condenser aperture diaphragm lever faces the lront.
When using a high NA condenser lens, screw the high NA
common condenser lens into the field l€ns section in the
diascopic illumination unit.
14

Chapter 4 Assembly
2.
3.
4.
5.
fl Wnen using phase contrast microscopy
lf you also wish to perform phase contrast microscopy, install the phase conlrast obiectives and the
condenser cassettes for phase contrasl observation that correspond to the condenser lenses to be used, as
described in the microscope instruction manual. Before beginning observation, be sure to center the annular
diaphragm. (Refer to the microscope instruction manual.)
The assembly procedure is now complete.
lf you require a very precise vibraîion direction adjustment, proceed with step 9.
lil Optical System Direction Adiustment (Adiustment of vibration direction)
To assure thal DIC images can be observed at optimum contrast, the vibration direction of lhe polarizer and
analyzer must be adjusted correctly.
1. Focus and center lhe system condenser as describ€d in the microscope instruction manual.
. Bring the A (empty) cassette into the optical path.
When using the condenser slider, remove the condenser cassette that is in the optical path so the
slider is empty.
. Bemove the polarizer, analyzer, and objective DIC prism from the optical path.
. When using a high NA condenser:
In order to focus the field aperture diaphragm image on the specimen surface, and in order to avoid
striking lhe spgcimen with the condenser lens, move the condenser lens slightly closer than the
subiect distance for each condenser lens, and then bring the specimen into focus while raising the
condonser lens. When using a oil-immersion type condenser, also check page g.
Remove the objective DIC prism that ls located directly below the 4Ox obiective.
Swilch to the 4Ox objective and bring the specimen into focus.
Move the stage so that the specimen moves oui of the viewfield. In its place, bring an almost dust-free
area into the viewfield. (There musî be no dust in the whole viewfield.)
Turn the Bertrand lens in/out lever to the [Bl position to bring the Bertrand lens into the optical path so
that the obiective's €xit pupil (the bright circle) can be observed. (lf your eyepiece tube has no Bertrand
lens in/out lever, install a cenlering telescope instead of an eyepiece and rotate its eyepiece part to
focus on the obiective's exit pupil.)
Vibration
dirgcton lgvgr
Vlbradon
dirgclion lgver
damp scf€r',
(bottom)
Figurc 4-9
6. When using the T-A2 analyzer, push in the analyzer to the
second clickstop so that it is in the optical path.
Loosen the clamp screw and align the index on the vibration
direction lever with the notch.
When using the epi-fl filter turret attached with the analyzer
block, place the analyzer block into the optical path.
Teble {-1
f .gmm (including slide glass with a thid(rless of f .2mm)
15

Chapter 4 Assembly
Whlte index mark
Figure 4-10
Da.k cross hat appears In the exit pupll
Figure /t-11
7. Push the DIC polarizer slider to the left so that the DIC
polarizer is in the optical path
Confirm that the white index mark on the DIC polarizer is
positioned at the center of the dial scale. (lf it is not, adjust it
as described in step g. of item S,.lnstalling the DIC polàrizer,"
on page 12.)
8, Loosen the condenser mount rotation clamp screw. Hold the
condenser turret and rotate both the condenser and the DIC
polarizer at the same time. Tighten the mount rotation clamo
screw wilh a hexagonal screwdriver to fix it at the Dosition
where.a dark cross* appears in the obiective,s exii pupil. Be
careful not to accidentally loosen the mount rotation ciamp
screw during DIC microscopy.
' The dark cross may not be visible with some objectives. In thts
case, rotate the eyepiece turet to the [O] positión, raise the
lamp voltage to the maximum, and then hold the condenser
turret and rotate both the condenser and the polarizer at the
same time while observing the specimen.
When the viewfield is at its darkest, tighten the mount rotation
clamp screw.
t
9. lnstall the .!0x objective's DIc prism that was removed in step
2. ln addition, if the condenser cassette in the opticat patn wls
removed from the condenser slider, install the condenser
cassette as it was originally.
The optical system's vibralion direction has now been properly
adjusted.
16

NF,:
Troubleshooting Tablek tililfl
Misuseoflheproductmayfesu|tinpoorproductpefformance,€veniftheproductisproper|yfunctiona|.|l
vJ"'"ipliiàié ànv or tn" rorro*ing'piour'".., check the folowing rabte for possible causes before
requesting service.
|f the prob|em is nol |isted below, or if the problem cannot be resolved by the suggested countermeasure,
unplug the power cord and conlact Nikon'
Viewfield vignetting
Contrast not obtained
when using the DIC
method
Poor viewing or contrast
Rotate to clickstop Position'
Botatg to clickstop Position.
Move the prism completely into the optical
oath.
Set to clickstop Position.
Install the revolving nosepiece on the
groove.
Inserî the polarizèr into ihe optical Path.
lnsert ths analyz6r inio the optical path'
Sèlsct a condenser cassette suitéd to the
condsnser lens being used. Sslect a
condgnser cassette suited to the
objgctive's indication
Move the prism complgtely into lhe
oplical path.
Use a Drism suited to the objective.
Datermine the crossed Nichols (dark cross)
position again.
Select a condenser cassette suited to the
condenser lens being used. Select a
condsnser cassette suited to lhe
obj€clive's indication.
Use a prism suited to the objscliv6.
Gently wiPe away tho dirt. (Becau6e this is
a mlarized intelgrence microscop€' dirt
Doses more of a problem lhan usual.)
While observing the odt Pupil ol the
obiective through the Bertrand lens' turn
ths revolving nosepièce slightly.
ll the bubble movgs with the nosepiEce, the
bubble is on the obisclive side.
ll the bubble remains stationary the bubble
is on the condenser side.
WiDe the immersion oil ffom the lens and
then apply it again'
. lf vou encoqntgf a problem other lhan thos€ disclss9d above, or if lhe action recommended in ihe labls above lails to conèct t,ìe
póblem, refsr io th€ mìcroscope instruction manual'
Optical path s\ritchover dial not in clickstop
oosiîion.
The condenser turrsl is in an intermediate
Dosition.
The objective DIC Prism is in an
intermediate Position.
The analyzer is in an intermediate Position'
Tha rgvolving nosepiece is not installed
properly.
The polarizer is not in the optical Path.
The analyzer is not in the optical Path
Wrong condensar cassette is s€lected.
The objeetive DIC prism is not in th€ optical
oath.
Wrong combination of objeclive and
obisctive DIC Prism.
Wrong condenaer direction.
Wrong condenser cassene is selectgd.
Wrong combination of obiectiw and
obiective DIC prism.
There is dirt on the objective, condenser, or
soeclmen,
Air bubble in the immersion oil on the lens'
17

Care and Maintenance
1. Filter and lens cleaning
Do not let dust, fingerprints, etc. get on the lenses or filters. Dirt on the lenses, filters, etc. will adversely
affect the view of the image. lf any of the lenses or filters get dirty, clean them as described betow.
' ' Use an air blower to blow away dust. lf that does not suffice, brush away the dust with a soft, clean
brush, or else wipe it away gen y with gauze.
' To remove fingerprints or grease, use a piece of soft, clean cotton cloth, lens lissue, or gauze
moistened with absolute alcohol (ethyl alcohol or methyl alcohol). Howèver, do not use ì-he same
area ot the cloth, etc. to wipe more than once.
' Use petroleum benzine to clean off immersion oil. Wiping with absolute alcohol (ethyl alcohol or
methyl alcohol) after the oil has been removed finishes the clean up process.
lf you cannot obtain petroleum benzine, use methyl alcohol. However, because methyl alcohol does
not clean as well as petrolEum benzine, it will be necessary to wipe the surfaces repeatedty. (Usually,
three or lour times is sufficient to clean lenses or filters.)
' Use petroleum benzine only to remove immersion oil from obiectives; do not use petroleum benzine
for cleaning the entrance lens on the eyepiece tube, filters, erc.
' Absolute alcohol and petroleum bénzine are both highly flammable. Handle them carefully, especially
around open flames and when tuming power switches on and off, etc.
' Follow the manufacturer's instructions when handling absolute alcohol and petroleum benzine.
2. Cleaning painted components
Do not use organic solvents such as alcohol, ether, or paint thinner on painted components, plastic
components or printed components. Doing so could result in discoloration or in peeiing of the printed
characters. For persistent dirt, dampen a piece of gauze with neutral detergent and wipe ligh y.
3. Storage
Store the product in a dry place where mold is not likely to form.
Store the obiectives, eyepieces, filter blocks, etc. in a desiccator or similar container with a drying agenî.
Put the vinyl cover overthe product after use to protect it from dust.
Before putting on the vinyl cover, tum off the power switches lor the microscope and the product used
together with the microscope, and wait until the lamphouse has cooled.
4. Regular inspection (charged)
Regular inspection (charged) of this product is recommended to maintain peak performance. Contacî
your nearest Nikon representative for details about regular inspection.
18