Nikon Labophot User manual

Nikon

Biological Microscope

INSTRUCTIONS

NIKON CORPORATION

CAUTIONS

oAvoid sharp knocks!

Handle the microscope gently, taking care

to avoid sharp knocks.

8When carrying the microscope

When carrying the microscope, hold its arm
with one hand, supporting the bottom of
the microscope base with the other. The
instrument weighs about 8 kg.

oPlace for using

Avoid the use of the microscope in a dusty
place, where it is subject to vibrations or
exposed to high temperatures, moisture or

direct sunlight.

e Light source

Usehalogen lamp 6V - 20W.

o In lighting the lamp

Take care not to touch the rear cover of
the lamp being lighted, and don't bring

inflammable substances such as gasoline,
thinner, and alcohol near to the cover, asit

may take a high temperature while the
lamp is being lighted.

o Focus knobs

Never attempt to adjust the tightness of
the right- and lefthand focus knob by turn-
ing the one, while holding the other in this
model microscope, because of causing

disorder.

o Exchanging the lamp bulb and fuse

Before replacing the lamp bulb (6V - 20W)
or fuse, turn OFF the power switch and
disconnect the plug of the power source
cord.
In such cases as of replacement, do not

touch the lamp bulb with bare hands,
immediately after putting out the lamp.

oDirt on the lens

Do not leave dust, dirt or finger marks on
the lens surfaces. They will prevent you
from clear observation of the specimen
image.

2

CARE AND MAINTENANCE

CONTENTS

oCleaning the lenses

T6 clean the lens surfaces, remove dust

using a soft brush or gauze. Only for
removing finger marks or grease, should
soft cotton cloth, lens tissue or gauze
Iightly moistened with absolute alcohol
(ethyle alcohol or methyl alcohol) be used.
For cleaning the objectives and immersion
oil use only xylene.
For cleaning the surface of the entrance
lens of the eyepiece tube and the prism

surface of the Trinocular Eyepiece Tube
"T" or the Ultra Wide Eyepiece Tube
"UW", useabsolute alcohol.
Observe sufficient caution in handling
alcohol and xylene.

f)Cleaning the painted surfaces

Avoid the use of any organic solvent (for
example, thinner, ether, alcohol, xylene
etc.) for cleaning the painted surfaces and
plastic parts of the instrument.

8Never attempt to dismantle!

Never attempt to dismantle the instrument
so as to avoid the possibility of impairing
the operational efficiency and accuracy.

NOMENCLATURE .

I.

II. ASSEMB LY .

MICROSCOPY .

III.

1. Operating Procedure .

2. Manipulation of Each Element

1) Interpupillary distance
adjustment .

2) Diopter adjustment .

3) Optical path change-over in the
trinocular eyepiece tube 9

4) Centering the condenser lens 10

5) Useof condenser aperture
diaphragm 10

6) Use of field diaphragm 11

7) Focusing 11

IV. OPTICAL SYSTEM .

V. PHOTOMICROGRAPHY 16
VI. USE OF THE ACCESSORIES 19
VII. TROUBLE SHOOTING TABLE 21

1. Optical 21

2. Manipulation 22

3. Electrical 23

4. Photomicrography 23

ELECTRIC SPECIFICATIONS

12

27

4

6

8
8

9

9

9

oWhen not in use

When not in use, cover the instrument with
the accessory vinyl cover, and store it in a
place free from moisture and fungus.

It is especially recommended that the
objectives and eyepieces be kept in an air-
tight container containing desiccant.

oPeriodical checking

To maintain the performance of the instru-
ment, we recommend to check the instru-
ment periodically. (For detai Is of this
check, contact our agency.)

3

I. NOMENCLATURE

Diopter ring

Interpupillary

distance scale

CF eyepiece

Eyeguard

R-revolving nosepiece

Stage Y-axis travel knob

Stage X-axis travel knob

Condenser focus knob

Daylight filter

CF objective

Stage clamp screw

Condenser aperture diaphragm

control ring

Condenser clamp screw

Filter receptacle

Brightness control dial

(Including power switch)

Dust cap

Fig. 1

4

Stage

Binocular eyepiece tube

Eyepiece tube clamp screw

Slide holder

Condenser

Condenser centering screw

Field lens

Field diaphragm control ring

Coarse focus knob

Fine focus knob

Lamp socket

Fuse holder

Power cord

Fig. 2

5

II. ASSEMBLY

• To assemble the microscope, follow the procedure in the order given:

Q)Leveling foot screw

For stable installation of the
microscope, manipulate the adjust-
ing screw at one foot on the bot-

tom of the microscope base.

L

J

®Lamp bulb

Insert the lamp bulb with its
pins into the accepting hole
in the socket.
Note: Don't touch the bulb

surface directly with the

fingers.

,

Pow" C",dl

Q) Lamp socket

Insert the socket into the
receptacle on the micro-

scope base.

/

I

!

iW

•

®Daylight filter

Place the filter on the field
lens. 45 mm in diameter.

6

-----------

®EyepieCe

Insert the eyepieces into the eye-
piece sleevesof the eyepiece tube.

/

@Eyepiece tube

After releasing the eyepiece tube clamp screw
arm, attach the eyepiece tube, and clamp it in

position.

I sufficiently on the right side of the microscope

(]) Objectives

Take stage down by manipulating coarse focus
knob. Mount the objectives on the revolving
nosepiece in such positions that, when, viewed
from the stage-side, their magnifying power in-

creasesclockwise ..

Slide holder

If the microscope is to be used with the lefthand
stage travel knobs, attach the slide holder to the
righthand hole, and if with the righthand knobs, to
the lefthand hole. Slide holder is to be removed

J

when using large specimen.

,,,j.- Stage clamp screw

Releasing the stage clamp screw suffi-

ciently, fit the stage into the circular

dovetail, and refasten the clamp screw.

Aperture number plate

®condenser

Insert the condenser into the condenser carrier,

facing the aperture number plate toward the
user, and fasten the clamp screw.

7

Fig.3

III. MICROSCOPY

I 1.~.,Op~rating~rocedure

1) Turn the brightness control dial (including power
switch) to ON and set the scaleon the dial to 4.

2) Remove the dust cap and place the daylight filter
onto the field lens.

3) Place the specimen on the stage and swing the

10x objective into position. Focus on specimen.

4) Adjust the interpupillary distance and diopter.
(Refer to P.9)

5) Carry out the centering procedure for the con-

denser. (Refer to P: 10)

6) Swi~g in the objective to be used and ~efocuson

specimen.

7) Adjust the condenser. (Refer to Table 1)

Table 1. Useof Condensers

\ Type of

20X

Remove the

Remove the condenser

condenser

condenser

condenser condenser

Abbe

Top lens swung in

Swing-out Achromat

2mm

NA = 1.25

Usable

Top lens swung out

NA = 1.35

1.8mm1,Gmm

Oil-immersion

NA = O,g

Achromat/aplanat

condenser

Dry system

Remove the

Usable

[NOTE] • The above object distance (from the top of the

condenser lens to the specimen surface) includes a
glassslide thickness 1.2mm.

*.When using the Swing-out condenser with 2X or

4X objective, fully open its aperture diaphragm.

• UW (ultra-wide) viewfield observation is possible
with 2X '" lOOX objective. In combination with
the Abbe condenser, however, the use of the lOX
or higher objective is possible.

• For photomicrography using the 4X or lower ob-

jective, remove the Abbe condenser.

• For photomicrography using the 2X objective,
preferably remove the condenser.

• For observation with the 1x objective, additional-
ly usethe diffuser. (available on order).

• The Achromat/aplanat condenser is not included
in the standard set.

8) Brightness is adjusted by changing the lamp
voltage.

9) Adjust the condenser aperture diaphragm and the
field diaphragm. (Refer to P. 10, 11)

8

2. Manipulation of Each Element

1) Interpupillary distanceadjustment

Place a specimen on the stage,and focus on the
specimen. As shown in Fig. 4, adjust the inter-
pupillary distance, so that both the right and

left viewfields become one.

2) Diopter adjustment

Make diopter adjustment for both the right and
lefthand eyepieces.

(1) Turn the diopter ring on each eyepiece,

until the end surface of the milled ring
coincides with the engraved line, as shown
in Fig. 5.

• Repeat the above procedure two times, and
a perfect diopter adjustment will be achiev-

ed.

• The above adjustment, compensating the
diopter difference between the user's right
and left eyes, will keep the tube length of

microscope correct, thus enabling him to
realize the full advantages of the highclass

objectives, including their parfocality.

(4) Since the CF eyepieces are of high eyepoint

type, it is not necessary for the user putting
on his spectacles to remove them.
Only fold down the eyeguard rubber.

(Fig.7)

Fig. 6

Fig. 7

Diopter ring

End surface

Engraved line

Fig. 5

(2) Mount the specimen on the stage. Swmg

the objective 40X into position, and bring
the specimen image into focus. For facilitat·

ing the focusing, first use the

lOX and then

40X objective.

(3) Thereupon, swing the objective 4X mto

position.
Without manipulating the coarse and fine
focus knob, turn the diopter rings on the
eyepieces, so that the specimen images in

the right and lefthand eyepieces are focus-
ed individually

3) Optical path change-over in the trinocular
eyepiece tube

(1) When using the trinocular eyepiece tube

As shown in Fig. 8, when the observation

tube is turned toward the user, 100% of

light enters the observation tube.

Observation
tube 100%

Fig. 8

As shown in Fig. 9, when the observation
tube is revolved 60° leftward, 100% of light
enters the vertical photo tube.

In either case,turn the tube to the limit.

9

Vertical photo
tube 100%

Fig. 9

(2) When using the trinocular eyepiece tube

"T" or the ultra wide eyepiece tube

"UW"

As shown in Fig. 10, with the change-over
knob pushed in, 100% of light enters the
observation tube.

Observation
tube

100"10

Optical path
change-over

knob

denser vertically so that a sharp image of
the field diaphragm is formed on the speci-

men surface.

(2) Bring the field diaphragm image to the

center of the field of view by means of the
condenser centering screws. (Fig. 12-W)

(3) Change over to the 40X objective, and

adjust the field diaphragm so that the

image of the diaphragm is about the same

as that of the field of view, as shown in Fig.

12-[Z]. If not centered, use the condenser

centering screws again.

Image of field
diaphragm

Eyepiece

viewfield stop

---

"",r Image of field

'\ diaphragm

Fig. 10

As shown in Fig. 11, with the change-over
knob drawn out, the proportion of light
entering the binocular observation tube
and vertical photo tube will be 14 : 86.

Observation tube:
vertical photo

tube = 14:86

Optical path
change-over

knob

Fig. 11

4) Centering the condenser lens

(1) Close the field diaphragm in the microscope

base to its smallest size by means of the
field diaphragm control ring. Rotate the

condenser focus knob to move the con-

Eyepiece

viewfield stop

Fig. 12

5) Use of condenser aperture diaphragm

The condenser aperture diaphragm is provided

for adjusting the numerical aperture (N.A.) of
the illuminating system of microscope. It is
important because it determines the resolution,

contrast and depth of focus.

In general, when it is stopped down to 70 ~
80% of the numerical aperture of the objective,
a good image of appropriate contrast will be
obtained. (Fig. 13)

Objective pupil

Aperture diaphragm

Size of the condenser aperture diaphragm

Fig. 13

10

The graduation on the Abbe condenser indicates
the diameters in mm of the aperture diaphragm

opening of condenser..
After removing the eyepiece from the eyepiece

tube, adjust the size of the diaphragm, observ-

ing the image of the diaphragm which is visible

on the bright circle of exit pupil of objective

inside.
It is recommended to take note of the diameter

of the diaphragm opening for each objective
power, whereby the best image is obtained.
The Swing-out Achromat and Achromat/
aplanat condensers, however,

indicating the numerical apertures (N.A.l. and

not the diameters of diaphragm opening.
Manipulation of these condensers isthe same as
that of the Abbe condenser. Stopping down the
aperture diaphragm too far will deteriorate the
image quality of microscope due to diffraction
of light. Therefore, it is not recommended to

stop down the aperture to a size smaller than

60% of the N.A. of the objective in useexcept

when observing almost transparent specimen.

have a graduation

the stage4.7mm.
The range of coarse and fine motion is within

30mm; 2mm up and 28mm down from the
standard position.
Tightness of the coarse-fine focus knob having
been properly adjusted by the manufacturer,
it should never be readjusted in this model
microscope by turning the one knob while
holding the other.

J

6) Use of field diaphragm
The field diaphragm is used for determining the
illuminated area on the specimen surface in
relation to the field of view of the microscope.

Generally, it is stopped down to such an extent
that the circumference of the illuminated area
circumscribes or inscribes that of the eyepiece

field of view. If the former be larger than the
latter, extraneous light will enter the field of
view, causing flare in the image and lowering

the contrast. Therefore, especially in photo-
micrography, the proper adjustment of the field

diaphragm is very important. Generally, good

results will be achieved when the diaphragm is
stopped down to such an extent that the dia-

meter of illuminated area is slightly larger than
the diagonal of film format.

7) Focusing

The relation between the direction of rotation

of the focus knobs and that of vertical move-
ment of the stage is as indicated in Fig. 14.

One rotation of the fine focus knob moves the

stage O.2mm.
The graduation on this focus knob is divided

2J.Lm.

into
One rotation of the coarse focus knob moves

11

IV. OPTICAL SYSTEM

The CF objectives and CF eyepieces adopted in
the Nikon Biological Microscope LABOPHOT
are designed on the basis of a new Nikon-
developed concept "Chromatic Aberration
Free" With the Nikon CF optical system the
chromatic difference of magnification in the
objective and eyepiece is individually corrected.

This is unlike conventional microscopes where
the corrections of such aberration has been, for

the most part, compensated for in the objectives
and eyepiece as a pair. As a result the Nikon
Microscope LABOPHOT has no orange colored
fringe in the eyepiece. In cooperation with the
other optimum aberration corrections such as
the Nikon Integrated Coating, a uniformly
sharp image, much superior in resolution,

contrast and color rendition is achieved over

100% of the effective, even, super-wide field of
view, for observation as well as color photo-
micrography.

.1. Objectives

Mechanical tube length of 160mm and parfocal

distance of 45mm (This is longer than the

33.6mm of earlier microscopes). In every case
use the CF objectives in combination with the
CF eyepieces.

1) Types of objective

(1) Achromat (CF)

In this type of objective, the correction of

chromatic aberrations is based on the lines
C (red) and F (blue). Importance being
given to the correction at the center of

viewfield, the objectives offer the finest
definition and highest contrast of image
at the center. Even the 40X and lOOX

objectives fulfill the "Chromatic Aberra-
tion Free" correction, wh ich has been
considered difficult so far until now for

such high magnifying powers. Furthermore,
image flatness has been attained to an

appreciable extent.

(2) Plan Achromat (CF Plan)

Same as the above type, the objectives ac-
complish the correction of chromatic

aberrations based on the lines C and F.
In addition, owing to sufficient correction
of all the image defects up to the periphery

of viewfield, the objectives provide an
unsurpassable high resolution and contrast

of image over a wider field.

Focusing at the center means simuItaneous
focusing at the marginal part of viewfield.

They are excellent for ultra-wide observa-

tion and photomicrography.

(3) Plan Apochromat (CF Plan Apo)

The use of fluorite and special, low color
dispersion optical glasses improves the

correction of chromatic aberrations over
the ent ire visible region up to the line g

(violet) along with the lines C and F.

These highest-grade objectives wi th their

large numerical apertures produce an ideal

image over a wide viewfield. With their
outstanding definition, superior color re-
producibility, and prominent image flat-
ness, they are especially suited for most

profound study of minute structures and
color photomicrography .

(4) Epi-fluorescence (CF UV-F)

Exclusively designed for episcopic, fluo-
rescence observat ion, th is type objectives
use non-fluorescent and non-solarisation
materials and a strictly chosen cementing
agent, to increase the transmission of UV
exciti ng light (u Itra-vi olet rays). Special
weight being attached to the correction at
the center of viewfield, and the numerical
apertures made extremely large, they
ensure bright and sharp fluorescence images
using every excitation method. As im-
mersion fluid, the objectives
of this type require the use of non-fluo-
rescent glycerine of high purity.

2) Useof the objective

(1) "Oil immersion objectives (Oil)

The objectives discriminated by the engrav-
ing "Oil" are to be immersed in oil between
the specimen and front of the objective.

When using oil immersion objectives of

numerical aperture 1.0 or higher, it is rec-
ommended, for making full use of its
efficiency, to use a highclass oil-immersion
condenser such as of Achromat/aplanat
type, applying oil between the glass slide

and condenser as well.

lOX ~ lOOX

12

To see if air bubbles are present in the
immersion oil, which deteriorate the image
quality, pullout the eyepiece from the

eyepiece tube to examine the

objective exit

pupil inside the tube.

remove air bubbles, revolve the nose-

To
piece slightly to and fro several times,

apply additional oil, or replace the oil.

Be careful not to rotate the nosepiece too
far as to soil the ends of the other objec-
tives with oil.

To clean off the oil, passlens tissue or soft
cloth moistened with xylene lightly two or

three times

over the lens. It is essential at

this time to avoid touching the lens with

the part of tissue or cloth once used.
Any remnants of oil left on the lens dete-

riorate the image quality.

(2) Coverglass

With the objectives engraved "160/0.17",

use a coverglass of O.17mm in thickness
(No. 1%). For the objectives whose N.A is

0.75 or higher, a coverglass of other thick-
ness than 0.17mm will deteriorate the

image definition and contrast.

The indication 160/- on the objective
means that no matter whether a coverglass

is used or not, no decrease of image defini-

tion or of contrast wi II resuIt.

(3) Objectives with compensation ring

When a high power, dry objective of large
N.A is adopted in combination with a
coverglass of thickness other than 0.17mm,
which will cause sharp reduction of image

definition and contrast, it is necessary to

use an objective incorporating a compensa-

tion ring asbelow:

First, observe with the compensation ring
set to 0.17, and then rotating the ring,
focus the image with the fine focus knob,
until an image of the highest sharpness and

contrast is obtained.

(4) No-coverglass objectives (NCG)

Objectives with the indication NCG are
suited for observing specimens such as
smearswithout coverglass.

(5) Objectives with aperture diaphragm

The objective incorporating an iris dia-
phragm serves to cut off direct light in

darkfield microscopy. Stop down the
diaphragm nearly to its minimum opening.

2, Eyepieces

To take full advantage of the CF eyepieces, use
them in combination with the CF objectives.
The indication "CF" should
their usewith other type objectives.

1) CFD eyepieces(CFD)

Being of wide field and high eyepoint type, the
CFD eyepieces are only used for observation,
obtains prominent image flatness. Compared
with the CFW eyepieces, they accomplish the
good correction of chromatic aberrations at the
periphery of the viewfield in combination with
the low magnifying power of CF Plan
Apochromat objectives.
They are equipped with a diopter ring and a
rubber eyeguard. An eyepiece CFD
incorporating a photo mask, is also available,

which enables focusing and framing by the use

of the observation tube of the Trinocular
Eyepiece Tube "T".

2) CFW eyepieces(CFW)

Being of wide field and high eyepoint type, the
CFW eyepieces with diopter ring are only used
for observation. They are equipped with a
rubber eyeguard.
An eyepiece called CFW lOX M, incorporating
a photo mask is also available, which enables
focusing and framing by the useof the observa-
tion tube of the Trinocular Eyepiece Tube "T".

3) CFUW eyepiece (CFUW)

Featuring extra-wide field of view and high
eyepoint, this eyepiece with diopter ring is
designed exclusively for observation. It enables
observation over a field of view twice as large as
that of the ordinary type eyepieces in combina-

tion with the ultra-wide tube.
An eyepiece called CF UW lOXM, incorporating

a photo mask, is also available, which enables
focusing and framing by the useof the observa-
tion tube of the Ultra Wide Eyepiece Tube

"UW".

4) CF PL Projection lenses(CF PL)

Exclusively designed for photomicrography. Do
not usethem for observation.

Every eyepiece is liable to gather dirt and dust,
which not only appear as shadows but also

impair image quality and contrast.

Keep the eyepieces clean at all times.

serve to prevent

lOXM,

13

3. Condensers

1) Abbe condenser

NA = 1.25. This is used with 4X - 100X ob-
jectives. The graduation of this condenser

indicates the diameters in mm of the aperture

diaphragm opening.

2) Swing-out Achromat condenser

= 0.9. Dry system.

N.A.

It is used in combination with objectives from
2X to 100X, and provided with a swing-out top
lens which is to be swung out when using the
2X or 4X objective. Its adjustable aperture scale

is graduated in N.A. ratings.

3) Achromat/aplanat condenser

NA

= 1.35. Oil system.

The spherical, coma and chrom<ltic aberrations
being ideally corrected, this large aperture
condenser is used with 20 X - 100 X objectives.
The standard thickness of glass slide should be

1.2mm.

Apply oil between the condenser and glassslide.

It is recommended that this condenser be

employed especially in combination with the
Plan Apochromat objectives. When using the

lOOX objective for observation in combination

with the CFW lOX eyepiece, it is possible to
close the field diaphragm down to 45% of the

viewfield.

4) Darkfield condenser(Oil)

= 1.43 - 1.20. Oil system. Used in dark-

N.A.
field microscopy. Apply oil between the
condenser and glass slide. (It is recommended
to usea thinner glassslide.)
This condenser is used in combination with the

objectives lOX - lOOX with aperture dia-
phragm (N.A.: up to 1.11.

5) Darkfield condenser(Dry)

= 0.95 - 0.8. Dry system. Used in dark-

N.A.
field microscopy. Magnifying powers of usable

objectives are lOX - 40X (N.A.: up to 0.7).

4. Illumination System (Fig. 15)

The optical system for illumination in the
LABOPHOT microscope is constructed to fulfill

the Koehler illumination requirements perfect-

ly, and offers a bright, uniform field without

any change-over manipulation.
As a standard light source, use the Halogen

lamp 6V 20W (PHI LIPS 7388).

OPTICAL PATH

Collector

lens

Halogen
lamp

Objective

Filter
Field lens

Fig. 15

14

(J1

5. Combinations of Objectives and Eyepieces

-

-

Depth 01 locus:

Glycerin

-

M

8x

16 x

32 X

32'x

M

M

.um

16x

With compen-

sal,on flng

80x

tJrnm

distance

Dowerlhlcknessndication

M¢mm

o 17

-

0.17

mm

2X(NAl' 7XN.A.xM

Real

Real

10

Real

dls1ance

nilicahon

locus

W.O.mmImm"m

¢ mm

¢mm

"m

o. I7

-

-

-

-

focus

viewtield

mfication

Depth 01

vlewfleld

WOfKlngFocusing

RealDepth of

Depth 01

v;ew'ield

nification

focus

viewfieldlocus

-

40 x

80x

"m

"m

Resolving

Total mag

Total mag

Total mag

non800x

non

lOx

n:Relractlve index of

DePTh of

Cove/glass

60x

1 8

0.45

0.3

non320x

50x

480x

non800X

u.

Total mag

4.5

1 7

800x

018

003

18

14

4.5

7 1

1 1

320x

11

1 7

1

0.45

09

1.8

1.4

160x

100x

0.9

4 1

045

2.8032

030.7

0306900x0.2305

04406

17

0.45

1 3

0343.2

400x

12

0.3611750x

31.0

5.3

4.1100x2 753

10013 90.28

320 x

1.1

1.0

0.8

06610

0.9012 9031

0.3

72

16.61.1

1.811.5

065

04

0.4

800x

05

0.450.8

07600x

06400x06607

0.085367 934

1.3

900x

26032

0.5

018

1500x

0.4

041500x

1000x

90155

9.0

30x

102

13.3

0164.733611 701732 x4 539

3360x3526

6.633

0403314.2069017

1 8

057 9042017160x0920

4 20.29

0180.4

0.18041500xo 14031000x

04

01804

04

0.3

02704

I 8108 79 2

18900

782

623

00')') 870. I5.5

90

25330x7.020520x13.3253

0113.8

28

4.572

6360x355140x6663

16 7

11 5

40x

0.45

0.4

09

1.3

1 2

09

017

47lOOx1.84.0

1.4

01780x1.869lOOx

1.44.4

160x

1 7

300x07

034

23200x

2.0

01

017

08

0.45

1 7

04

0180.4

03

02

0.1

017

0180.5

1.6

wavelength I

022

0.4

0.4

017

12

10

0.661.2

1.8

200x1 31.8

10.1

8.2

2 710 I

84069

160x0.9

2 9

09400x0.45

035o

3.5300x0.72.9200x1.3

048

0.42

320x0451.3

1.2

10400x0.661 2

035

600x

600x

0.45

1.6

035

032017

036

500x

0.2809500x0531.1

20

28

150x1.4

017

045

400x

600x035

400x

0.7600x

07900X

0.440.7

40X456360x

56

lOOX1.810.1150X1.48.2

4.4

1 25

018

1500X0.14

022

0.18

0.35

0.45

0.30.7

0.307

0.2306600x0440.7

0.85

0.18

1000x

05

014

1000x0.27

021.80.220.17800X0.18051000x018

0.1404

0.270.4

132

7 .0

20x

132

40x

40x

80x

6 2

065

02o 17

027

1000x

1500x0.14

1000x

lOX

289

9.0

4011

40x

0.2')

1.8

0.65

4.2042

228.80.69

4.0

0.93.5300x072 9

045

400X0.45

600x0.3510

134005

0.55

80x1.8

150x

30

0.28162

1.85.8

7.80.37017

0.9

200x091.4

20x

8 9

0.17160x09

0.9

300x0.71 5

1.3

320x045

400X

0.6600x03505lOa x1.3

0.210.17

1000x

1500X0.14

0.9

031

800x0.18051000x

1500x0.140.41000x0270.5

0.31

1.8

017800Xo 18051000x018

1500x0.14

Wdh compen-

600x0.35

400x

300x0.71.4

150x1.4

100x

04

017

40

8.80690.17

0.940200x0.93.5300x0.7

0.374.20320.17320x

08600x

3 5

065

017

600x0.35

085

600X

0.85

0.1

0.2306600x

45

3.551025

053

0420.17

0.14

017

400x

600x

0.43

05

20X

200x

1000x

20x

lOOX

200X

.14.3

0.55

150X

0.75066

0.802

40x

4.50.21

013

800X018

1 25014

200x

04223

085

045

400x

400X

05

lOa x

055

05

480 x

0.,9:..\7,,,480x

320 X

0

::>

320x

160x

nxA

(Resolving power at eye=2'

Resolving power: __ A__

CF Objectives (160/45)

-

NCG

1

1 3')o 17

7

0.45

Magnification

0,1

15 x

0.90.26

1.25

20 x

I 3')

60x

40x

20x

100x

+ n

100x

20x

aperTure

40x

2XNA

Field number= 18

CFW15X

Field number= 26.5

CFW lOX

CFUW lOX

40 x095 400x

0,1

FieId number.-14

60x

01

1.8

1011- 0 23)

1000x

10- 2.0)

10- 2.01

Ultra-Wide vlewfleld

CFW8x

( A= O.5511mSianda'd

Ordinary viewfield

CF Eyepieces

Table 2

object side)

v .PHOTOMICROGRAPHY

(The Biological Microscope LABOPHOT IS

designed mainly for observation.)

1. Combination of CF Objectives and
CF

PL Projection lens

The combined use of the CF objectives and CF

PL Projection lens is essential.

For the same total magnification, select a

combination of the highest possible objective

power and lowest possible projection lens
power to achieve the utmost image definition

and contrast.

2. Checking the Illumination

Uneveness in the illumination will show up
more conspicuously in photomicrography than
in observation. Consequently, before taking a
photograph, recheck the correct adjustment of

the condenser.

3. Selection of Voltage and Filter

1) When using a daylight type color fil m

Set the brightness control dial to 5.5 and use
the NCB 10 fi Iter

Adjustment of the image brightness should be
made by meansof the NO filters.

2) When using a monochrome film

Remove the NCB 10 fi Iter. Contrast fi Iters such
as X-1 green are usable.

*-' The NCB 10 filter is most suitable for a

standard film. Depending upon the make
of the film different color renditions may
result. It is recommended that in addition
to the NCB 10 filter a color compensation
filter (CC filter), available from the film
manufacturer, be used.

*-'

"UW" tube

"F" tube

or

tube

finder

eyepiece

"T"

Use

Use

Failure" of film may result. So, when taking
picture of such specimens, it is recommended
to use the Nikon Biological microscope OPTI-

PHOTo

5. Manipulation of Field and Aperture

Diaphragms

In photomicrography, the adjustment of the
field diaphragm is important for the purpose of

limiting extraneous light which causes flare in
the microscope image. Stop down the dia-
phragm so as to get an illuminated area slightly
larger than that of the picture field. By adjust-
ing the aperture diaphragm, a change of depth
of focus, contrast and resolution of image is
attainable. Select a size suited to the purpose.
Generally speaking, the aperture diaphragm, is
properly stopped down to 70 ~ 80% of the
aperture of the objective being used.

6. With Regardto Condensers

For photomicrography, it is generally rec-

ommended to use the Swing-out Achromat
condenser. When using 2X objective, however,
preferab Iy remove the condenser.

7. Focusing

Focusing is to be accomplished by means of the
ocular finder on the photomicrographic attach-

ment, or binocular observation tube with mask
eyepiece on the trinocular eyepiece tube.

Table 3. Focusing

observation

4 x or

Ocular

Use

Ocular

10x or

Type of

or

finder

Use

+ magnifier

fi nder

lower objective

Focusing with

tubeOcular

observation

higher objective

Focusing with

Focusing

tube

+ magnifier

Focusing

4. Shutter Speed

Desirable shutter speeds for least vibration are

1/4~ 1/15sec.
Adjustment of the image brightness for color
photomicrography should be made by means of

the NO filters. Some specimens require, on
account of their insufficient brightness, longer
exposure times, and consequently poor color

reproducibility owing to the "Reciprocity Law

16

CD Adjust diopter.

• Binocular of eyepiece tube:

Use4X or lOx objective.

Insert the mask eyepiece into either of
right or left eyepiece sleeve that is ac-
customed to usual use. Adjust the dioPter
ring to bring the double cross line in the

view field center into focus. (Fig. 16)

Then focus the specimen image also on the
central area of the mask by means of the
focus knoq of the microscope.

The diopter of another eyepiece is to be
adjusted by focusing specimen rotating the
diopter ring without using the microscope

focus knob.

Rotate the mask eyepiece so as the mask

positions asshown in Fig. 19.

• Ocular finder:

Adjust the diopter ring so as the double
cross line in the

view field center can be

seenclear and each line separated. (Fig. 17)

Double cross line ::::::mE''':;''' __
of the mask eyepiece II II

Double cross line «; <» ••• ~ ~

of the ocu lar finder Af;7 ~ ~ ~

111 __ II

Fig. 16

the double cross line so as their relative
position is fixed and unchaned under

observation by swinging your eye laterally.
(Focusing by parallax method.)

• Using 4X or lower objective:

Attach the focusing magnifier to the
ocular finder. (Fig. 18)

Focusing magnifier

Ocular finder

\lilill

Viewing through the attached focusing
magnifier, move it back and forth until the

double cross line is seen clear. Then, focus
the double cross line and the specimen
image by rotating the fine focus knob as
sharp as possible.

8. Picture composing

Compose the picture within the mask in
the ocular finder corresponding to the film
size in use by driving the microscope stage

by lateral and longitudinal

rotation. (Fig. 19)

movement and

o

Fig. 18

Fig. 17

(2) Make focusing according to the magnifica-

tion of objective to be used.

• Using 40X or higher objective:

With diopter adjusted eyepiece make the
specimen image sharp by rotating the
microscope fine focus knob and make sure

that both of the double cross line and the
specimen image are seen crisply at the same

time.

• Using medium magnification objective
10X, 20X, etc.:

After focusing the same way as

above,

bring the specimen image to coincide with

17

For 3Smmfilm

For 4"XS"
Polaroid film

ForJ~'X4~'
Polaroid film

For6X9
roll film

Double crossline

Finder mask

Fig.19

Masks of the mask eyepiece are appli-
cable to 35mm film picturE! composing .

Select one out of masks suitable to the
CF PL Projection lens in use.

Note that, however, framing for picture
composing will be more accurate by the

ocular finder than the mask eyepiece.

Inner frame

(for CF PL5 X)

Intermediate

frame

(forCF PL4X)

Outer frame

(forCF PL2.5X)

Mask of the mask eve piece

Fig. 20

9. Vibration-free operation

Set the microscope on a vibration-resistant.
rigid desk or a bench with a vibration-proof

device.

10. Others

• When using the 1 X objective. place the
diffuser (available on order), and remove
the condenser.

• For photomicrography, when focusing
with the binocular observation tube, use
the CF eyepiece, CF PL Projection lens and

CF photo mask eyepiece. with the magnifi-
cation and other indications engraved in
yellow, or in white with a white dot in
addition.

• For the use of other photomicrographic
attachment refer to the perti nent instruc-

tion manuals.

18

VI. USE OF THE ACCESSORIES

1. Ultra Wide Field Trinocular Eyepiece
Tube "UW"

1) Objectives

CF Plan Achromat 2X'" lOOX, CF Plan Apo-
chromat 2X'" lOOX, CF Plan Achromat for

phase contrast 1OX'" lOOX, CF Plan Achromat
for metallurgical 5X '" lOOX, CF Plan Apo-
chromat for metallurgical 50X or CF SO Plan
Achromat for bright and darkfield 5X'" 1OOX

are used.

2) Condenser

Refer to the Table 1 (P.8).

3) Assembly and microscopy

Assembly and microscopy being almost the
same as that of the regular microscopy (P. 6
and P. 8). only the differences will be described

below.

(1) Using the centering telescope

For attaching the centering telescope on

top of the eyepiece sleeve, it is necessary to
use the adapter (Fig. 21), because the tele-

scope which has been originally designed
for centering the annular diaphragm in
phase contrast microscopy, has a fitti ng

diameter different from that of the CFUW

eyepiece.

2. Polarizing Filter Set "PT"

1) Nomenclature (Fig. 22)

Analyzer

Polarizer

2) Assembly

(1) Attaching the analyzer

After removing the eyepiece tube, insert
the analyzer into the optical path hole in
the microscope arm. (Fig. 23)

The white index dot is to be brought into
coincidence with the Y-axis (of X-V co-

ordinates), viewing the arm from above.

"

White ~y

indexdot~AnaIYZ~r

•

Fig. 22

Fig. 21

Fig. 23

(2) Condenser

Usethe Swing-out condenser.

(3) Attaching the polarizer

As shown in Fig. 24, fit the polarizer to the

internal diameter at the bottom of the

condenser.

t

S--POlarizer

Fig. 24

19

(4) Objective

Usethe ordinary CF objectives.

3) Microscopy

(1) Turn ON the power switch. Set the bright-

nesscontrol dial to 4.

(2) Remove the dust cap and place the daylight

filter.

(3) Place the specimen on the stage and focus

on specimen with 10Xobjective.

(4) Adjust the interpupi Ilary distance and

diopter. (Refer to P. 9)

(5) Swing in the top lens of the swing-out

condenser in the optical path. (If using 4X

objective swing out the top lens.)
(6) Center the condenser. (Refer to P. 10)
(7) Rotate the polarizer until the darkest field

view is obtained.

of
(8) Set the brightness control dial to 5 - 6.

(9) Change

over the objective to be used and

sharpen the focus on the specimen.

(10) Adjust the apertu re diaphragm and field

diaphragm. (Refer to P. 10 and 11)

(NOTE)
The following accessories can not be used in

combination with LABOPHOT (Y-R stand)
Microscope.
eTeaching Head and Multi-teaching Head

(Only when they are combined with Ultra

Wide Eyepiece Tube "UW")

eEpi-iliuminator "M"

20

VII. TROUBLE SHOOTING TABLE

Although nowhere the user can find any disorder or derangement in the instrument, if he
encountes some difficulty or dissatisfaction, recheck the use, referring to the table below:

1. Optical

field

resolution or

(No appearance
Image quality

)Actions

• Dirt or dust on the lens

• No coverglass attached to slide or

• Condenser not centered ) Centering by using field dia-

• Too thick or thin coverglass

• Condenser aperture too much closed

Failures

• Dirt or dust on the entrance lens

(Refer to P. 10)

(Refer to P 10)

• Dirt or dust on the lens

• No immersion oil used on immersion---+ Use immersion oil

• Objective aperture (which provided)

field diaphragm image

• Dirt or dust on the slide

• Improper use of condenser

system objective

too much opened

• Revolving nosepiece not in click-

• Revolving nosepiece not in c1ick-

• Too low position of condenser

• Optical path in trinocular tube not

Causes

centered in optical path)

(Condenser, objective, eyepiece, field lens)

NCG objective used with coverglass

stop position (Objective not

stop position

fu Ily changed-over

(Refer to P. 9)

, Correct use (Refer to P. 10)

) Revolve it to click-stop position

) Remove bubbles

) Changing-over to the limit

) Cleaning

) Cleaning

) Use Nikon immersion oil

) Revolve it to click-stop position

' Cleaning

) Correct positioning

(Refer to P. 12)

) Correct use (Refer to P. 13)

) Open properly (Refer to P. 10)

) Use specified thickness (0.17mm)

) Cleaning

) Open it properly

, Cleaning

) Bring it up to coincidence with

) Cleaning

) Adjustment (Refer to P. 13)

) Open properly

ph ragm (Refer to P. 10)

coverglass (Refer to P. 13)

21

Failures

• Condenser not correctly centered

Actions

• Specimen rises from stagesurface

• Revolving nosepiece not in c1ick-

• Daylight fi Iter not used

Causes

stop position

2. Manipulation

.

• Place it stable

, Revolve it to click-stop position

(Refer to P.9)

• Use daylight filter

• Correct centering (Refer to P.l0)

,Changing-over to the limit

Failures

slide

slide

smooth by

(when changed-over)

moving the

Movement of

No fusion of

High power ob-

Travel of stage

Fatigue of

Actions

Causes

.

holder

• Incorrect diopter adjustment

• Upside down of slide

• Upside down of slide

• Too thick coverglass ' Use specified thickness (O.17mm)

• Eyepiece diopter not adjusted' Diopter adjustment

• Slide holder not tightly fixed

• Eyepiece diopter not adjusted• Diopter adjustment

• Improper attaching of slide

• Interpupillary distance not

adjusted

• Too thick coverglass

(Especially when changing-over

• Inadequate brightness of illumination~ Change power voltage

' Turn over the slide

• Turn over the slide

(Refer to P 9)

• Use specified thickness (O.17mm)

) Fix it tightly

) Adjustment (Refer to P. 9)

) Correct adjustment(Refer to P.9)

) Shift the attaching position

(Refer to P 9)

coverglass (Refer to P. 13)

coverglass (Refer to P. 13)

22

3. Electrical

illumination

Failures

bulb (PH ILIPS 7388)

(Refer to P. 10)

• House current voltage fluctuates

• Lamp bulb insufficiently inserted

• Too low voltage

Actions

• Fuse blown

• Lamp bulb going to be blown

• Lamp bulb not inserted to the limit

• No lamp bulb attached • Attaching

• Lamp bulb blown ) Replacement

• Condenser not centered

• Not specified fuse used ) Use 1A (250V) or O.5A (250V)

• Fuse holder not firmly fastened

• Irregular change of house current

• Input voltage not adjusted to

• Not specified lamp bulb used

• Condenser aperture too much c1osed---. Open it properly (Refer to P. 10)

• Too low position of condenser

• No electricity obtained

• Too high voltage of house current

Causes

house current voltage (for Europeanthe microscope bottom

districts only)

•

• Secure connection

• Positive connection

• Usetransformer or the like

• Connect the cord to socket

• Use stabilizer

(for adequate voltage)

) Use transformer for adjustment

) Centering (Refer to P. 10)

• Replacement

) Turn the change-over switch on

I Use6V 20W specified lamp bulb:

• Correct positioning

• Replacement

• Use 6V 20W specified Halogen

• Cleaning

• Raise the voltage

• Firm fastening

(Halogen bulb: PHILIPS 7388)

picture

4. Photom icrography

Failures

crosshair.

addition.

separation appears between the image and double

• At lower magnifications use focusing telescope in

.· Actions

Causes

• Improper focusing----. • Viewing into the finder and turning diopter ring,

laterally, rotate fine focus knob, until no parallax

bring double crosshair into focus. Moving the eye

23

)

Failures

• When contrast is to be increased for a part stained

more conspicuously

in observation)

contrast.

istics will be unavoidable.

white film).

black·and·white film).Note. however. for color film. that lowering of

in photography than

(Refer to P. 16)

chromatic interference filter (e.g. peak wavelength= 546nm. half-value range = 30 nm) will increase

color temperature and change of spectral character-

with a particular color. use a filter whose color iscomplementary to the stain color (for black-and-

• Using dry objectives---+. Use no·coverglass type objective.

on the specimen.

condenser lens. field lens. etc.

field diaphragm

for smear preparations

of coverglass

Actions

centered

• Out of focus

• Grease. dust or dirt----+. Clean the front of objective thoroughly. top surface

• Condenser not

• Aperture diaphragm---+. Generally. good resu Its wi II be achieved with

(No.1 %)

Causes

on optical surfaces

• Use objective With coverglass thickness compensa-

(This shows up

opened too large

of projection lens. specimen. photographic lens.

contrast microscopy. use of a green fi Iter or mono-

(Especially with high

• Select a place free from vibrations. such as caused

). Centering (Refer to P. 10)

tion ring.

aperture stopped down to 70 - 80% of N.A. of the

• If other objectives are to be used. place a coverglass

) • For preventing external vibration. use vibration-

proof table or rigid desk.

power objective and

by traffic. passers-by or motors etc.

larger than the diagonal of picture frame.

). In metallurgical. interference. polarizing or phase

objective being used. (Refer to P. 10)

(for color film. to 1/4 -1/15 sec.)• Lower the voltage. and elongate exposure time (for

24

obtained

make or emulsion NO.

film development

source voltage

flare

resolving

graph

fi nder

Actions

)

filter

• Film of another

• Extraneous light~. Darken the surroundings or place the cap on the

Failures Causes

• In color photography, depending upon the speci-

• Incorrect exposure---->. By inadequate exposure time, color rendition willtime

of field is reduced.

• Specimens should be stained a rather dark color, if

• Even though of same make, according to emulsionnumber, different color rendition will be obtained.

• Stray Iight entering

• In black-and-white photography, for low contrast

is more suited (such as minicopy film).

cation

Then, with the help of exposure time indicator,adjust exposure time according to characteristics offilm by means of NO filters, or compensate forsuch fai Iure by means of CC fi Iters.(Refer to Kodak Data)

men, red-blue separation staining (Mallory or Azanmethods etc.) is preferable to red-violet combina-tion staining (H-E staining).

specimens a film of finer grain and higher contrast

• For general specimens a film of wider latitude and

upon the type, make, etc.

finer grain is preferable.

possible.

entering the ocularocular finder.

• Low contrasT in

• Inadequate useof-----. Select best filter combination.

• Insufficient N.A.~. Usea large N.A. objective.

-----+. Take care not to expose microscope and specimen

contrast. darkfield, or differential interference

resolving power.

of objective

• • Take picture in every caseat the specified voltage.

). To increase contrast optically, select phase

not be true on account of "reciprocity law failure"

• For the same magnification, increase power of

•• Note that, when using a daylight film, remarkably

different spectral sensitivities will result depending

specimen

mended to contact the development laboratory.

) • Especially, for making color prints, it is recom-

to direct sunlight and other intense lights.

higher resolution and sharpness, even though depth

methods.

objective rather than that of eyepiece to attain

(Refer to P. 16)

25

Power source

Halogen lamp

220/240V

ELECTRIC SPECIFICATIONS

6V 20W

100V

120V

50/60 Hz

0.5A (250V)

220/240V

(PH ILIPS 7388)

100V}

1A (250V)

120V

Nikon reserves the right to make such altera-

tions in design as may be considered necessary
in the light of experience. For this reason.
particulars and illustrations in this handbook may
not conform in every detail to models in current

production.

27

NIKON CORPORATION

FUJI BUILDING 2-3. MARUNOUCHI 3-CHOME,CHIYODA-KU, TOKYO 100,JAPAN
PHONE: [phone] TELEX: J22601 NIKON, FAX: [phone]

Printed in Japan

6(89.2.B)H· E-6