Nikon Eclipse Ti Series User manual

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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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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