Nikon DIAPHOT 300 User manual

Ill
Instructions
NIKON CORPORATION

Thank you for your purchase of Nikon’s Inverted
Microscope Diaphot 300. Please read this instruction
manual thoroughly in order to become acquainted with the
complete system and its operation. We hope your Diaphot
300 will be of lasting service.
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.

Handling Precautions
1. Handle Carefully!
Handle the microscope gently, taking care to avoid sharp impacts.
2. Purpose
Use the microscope only for microscopic observation. Do not use it for
any other applications.
3. Microscope Location
Select a location with limited exposure to dust, vibration, high temperature
and humidity (40°C, 80% or more), and direct sunlight. Leave a certain
space between the microscope and the nearby wall to allow the user to
look at the warning labels on the lamphouse.
4. 12V100W Power Supply Unit
The power supply unit specified on page 53 "ELECTRICAL
SPECIFICATIONS" is the power supply unit to light up the Nikon
microscopes' 1 2V100W lamp. Do not use it for other purposes.
Also, the user may not use the power supply unit if it is broken, and may
not open the cover of the power supply unit.
5. Line Voltage
Confirm that the input voltage to the 1 2V1 00W power supply unit
corresponds to your line voltage. Be sure to use the specified power
supply unit (described in page 53 "ELECTRICAL SPECIFICATIONS").
6. Light Source
The microscope uses a 1 2V, 1OOW halogen lamp bulb as the standard
light source. Do not use any lamp not described in the "ELECTRICAL
SPECIFICATIONS" (on page 53). If a lamp bulb of over-rated wattage is
used, the light adjusting circuit may be damaged.
7. With Lamp On —CAUTION —
Do not touch the lamphouse or place any heat-sensitive object near it,
since the lamphouse becomes extremely hot during use. Also use great
care not to bring volatile substances (such as gasoline, thinner, alcohol,
etc.) close to the lamphouse. Such exposure may result in inflammation
or other dangers.
8. Replacing the Halogen Lamp
Before replacing the 12V, 100W halogen lamp, turn off the power switch
and unplug the power cord. When replacing the halogen lamp, wait until
it cools down. Do not touch the glass of the halogen lamp with bare
hands.

9. Dirt on Lens
Do not leave dust, dirt or fingerprints on the lens or lamp. Dirt or stains
on the lens or mirror will deteriorate image quality.
10. Focusing Knobs
Never attempt to turn the right-hand and left-hand focusing knob in
opposite directions at the same time as it will result in defects or damages.
An attempt to turn the coarse focusing knob further after it reaches its limit
will cause defects or damages. Never turn it forcedly.

Care and Maintenance
1. Lens Cleaning
Dust is best removed with a soft brush or gauze. More persistent dirt,
such as fingerprints, grease and oil, may be removed with soft cotton,
lens tissue, or gauze lightly moistened with absolute alcohol (methyl
alcohol or ethyl alcohol).
Use only xylene to clean immersion oil off objective surfaces.
Do not use xylene to clean the entrance lens at the bottom of the eyepiece
tube or prism surface of the eyepiece tube.
Absolute alcohol and xylene are quite inflammable. Use great care when
handling them and when setting the power switch on and off. Be very
careful with fire.
2. Cleaning Painted Surfaces
Avoid use of any organic solvents (such as alcohol, ether, thinner, etc.) to
clean the painted or plastic surfaces of the instrument. We recommend
that these parts be cleaned with silicon cloth.
3. Never Dismantle
Never attempt to dismantle the instrument, thereby avoiding the possibility
of impaired operational efficiency and accuracy.
4. When Not In Use
When the microscope is not in use, cover it with the vinyl cover, and store
it in a dry place not subject to mold.
We especially recommend that the objectives and eyepieces be kept in a
container (such as a desiccator) with desiccant in it.
5. Periodical Inspection
To maintain the performance of the microscope, we recommend you to
check the microscope periodically. (For details of inspection, contact your
dealer or nearest Nikon representative.)

I. NOMENCLATURE
CONTENTS
Handling Precautions
Care and Maintenance
I . NOMENCLATURE. 1
II. BASIC MICROSCOPY. 4
III. PHOTOMICROGRAPHIC PROCEDURE. 1 2
IV. OPERATIONS ON COMPONENTS . 16
1 . ON/OFF switching of power. 1 6
2. Adjusting Brightness.17
3. Changing Optical Paths.19
4. Filters.20
5. Field Aperture Diaphragm . 21
6. Condenser Aperture Diaphragm . 22
7. System Condenser Turret Unit . 24
8. Objectives.26
9. Turret Assembly.28
10. Diopter Adjustment.29
1 1 . Adjusting Interpupillary Distance .30
1 2. Photomask.31
1 3. Eyepiece tube.32
14. Focusing Device.33
15. Refocusing Stopper . 34
16. Handling Condenser Flolder . 35
17. Tiltable Illumination Pillar.36
18. Rectangular Mechanical Stage.38
19. Photomicrography.39
V. ASSEMBLY . 4 2
VI. SYSTEM DIAGRAM . 4 8
VII. TROUBLESHOOTING . 4 9
• ELECTRICAL SPECIFICATIONS . 5 3
II. BASIC MICROSCOPY
III. PHOTOMICROGRAPHIC
PROCEDURE
IV. OPERATIONS ON
COMPONENTS
V ACCPMRIV
V , MOO Cl VIDL I
VI. SYSTEM DIAGRAM
YU. TROUBLESHOOTING
• ELECTRICAL
SPECIFICATIONS

I. NOMENCLATURE
1. System Components (Right-hand)
Supplementary lens pocket
Field aperture diaphragm lever
Figure 1-1

2. System Components (Left-hand)
Condenser
refocusing stopper
Condenser mount rotation clamp screw
Condenser holder
Module
Condenser turret
Annular diaphragm
centering screw
Condenser lens
Objective
Revolving
nosepiece
Side port
Coaxial coarse/fine
focusing knob
Refocusing stopper
Light intensity control dial
Condenser mount
Condenser centering
screw
System condenser
turret unit
Bertrand lens
focusing screw
Eyepiece tube port
Front port
Figure 1 -2
2

3. System Components (Rear)
Lamphouse cable connector
Figure 1-3
3

II. BASIC MICROSCOPY
For the normal microscopy, carry out steps 1 . to 14. For phase contrast
microscopy, carry out steps 1 . to 18. For detailed operations, see Section
IV. "OPERATIONS ON COMPONENTS ", from page 16.
If the microscope has not been assembled, first read Section V . "ASSEMBLY" from
page 42.
1. T urn on the power.
After confirming that the power supply
is set for the correct line voltage, turn
on the power switch. Set the light
intensity control dial on the power
supply unit to CTRL.
Figure 2-1
2. S et the lamp voltage to 6.
Rotate the light intensity control dial on
the left side of the base to the "6"
position. (If the light intensity is too
high or too low, adjust it properly.)
3. S et the optical path for the observation.
Set the turret assembly to 0. (Eyepiece
tube BT1 only.)
Set the optical path changeover dial on
the right side of the base to A.
Figure 2-3

4. S elect the NCB11 filter.
Hold the nonslip portion of the filter
slider to push in the NCB1 1 filter. Do
not touch the filter. Keep other filters
out of the optical path.
For phase contrast microscopy, set the
GIF (green interference) filter, instead of
the NCB1 1 filter, in the optical path.
Figure 2-4
5. F u ll y open the field aperture and
condenser aperture diaphragms.
Raise the field aperture diaphragm
lever.
Turn the condenser aperture diaphragm
lever to the right.
6. A djust the condenser position.
Adjust the condenser to an
approximately correct position.
• System condenser turret unit
♦ LWD condenser
Rotate the condenser refocusing
stopper by half turn to loosen it,
and rotate the condenser focus
knob to lower the condenser to its
limit. Rotate the turret until the module A comes to the front (i.e., it enters
the optical path).
♦ ELWD condenser
Rotate the condenser focus knob to raise the condenser to the upper limit,
then lower the condenser by approx. 1cm from the limit. Rotate the turret
until the module A comes to the front (i.e., it enters the optical path).
• ELWD condenser
Rotate the condenser focus knob to raise the condenser to the upper limit,
then lower the condenser by approx. 2cm from the limit.
• SLWD condenser
The SLWD condenser requires no adjustment since it is fixed.

7. M o ve the 10 x objective into
the optical path.
Move the 10x objective into the optical
path by rotating the revolving nosepiece
securely into the clickstop.
8. A djust the eyepiece diopter.
Turn down the reticle in/out lever to the
right to bring the photomask into the
optical path.
Rotate the diopter adjustment ring on
each eyepiece to bring the photomask's
double crosshairs into sharp focus. (Be
sure to perform this step for each eye,
as correct focus for each eye is usually
different.)
If the optional rubber eye guards are
used and if you wear glasses, fold
down the rubber eyeguards.
9. A djust the interpupillary distance.
Adjust the interpupillary distance until
the viewfield is visible as a single image
through both eyepieces.
Figure 2-9

10. Place a specimen on the stage.
If the specimen is on a slide glass, be
sure to mount it with its cover glass
facing down. (See page 27.) Move
the stage to position the desired section
of the specimen into the viewfield.
Figure 2-10
1 1. Focus.
Rotate the refocusing stopper on the
left-hand focusing knob counter¬
clockwise to unlock the stopper.
Rotate the coarse/fine focusing knob to
focus on the specimen.
NOTE: Never turn the left-hand and
right-hand knobs in the
opposite directions at the same
time, as damage may result.
12. Center the condenser.
Narrow down the field aperture
diaphragm until the diaphragm edge is
viewed through eyepieces.
Rotate the condenser focus knob to
focus the image of the field aperture
diaphragm on the specimen surface.
Adjust the condenser centering screws
so that the center of the field aperture
diaphragm image matches the center of
the viewfield.
NOTE: If the SLWD condenser is used,
this centering is unnecessary.
(Fully open the field aperture
diaphragm.)
Figure 2-12
Figure 2-1 1
7

13. Switch to the 40 x objective
and re-center the condenser.
Switch to the 40 x objective.
Adjust the image of the field aperture
diaphragm until it is circumscribed with
the viewfield.
If the center of the field aperture
diaphragm image is off the center of the
viewfield (eccentric condition), readjust
with the condenser centering screws.
If the LWD condenser is used, tighten
the condenser refocusing stopper.
14. Perform microscopy with a
desired objective
♦ Switch to a desired objective.
♦ Focus the microscope.
♦ Adjust the field aperture diaphragm
until it is inscribed or circumscribed
with the viewfield.
♦ Adjust the brightness with the ND
filters or the light intensity control dial
on the base. (Maintain the lamp
voltage between 6 and 12. For color
photomicrography, adjust it to
PHOTO. See page 17.)
♦ Stop down the condenser aperture
diaphragm to a range of 70 to 80% of
the objective's numerical aperture.
(See page 22.)
♦ When replacing the specimen, you
can use the refocusing stopper (page
34), condenser refocusing stopper
(page 35), and pillar tilting
mechanism (page 36) for enhanced
efficiency.
Now, the basic microscopic operation is completed.
Improved image quality may be achieved by slightly opening or closing the
condenser aperture diaphragm. (See page 22.)
For phase contrast microscopy, proceed with the following steps 15. to 18.

Phase Contrast Microscopy
1 5. Switch to the phase contrast
objective.
Rotate the revolving nosepiece to bring
the phase contrast objective of the
lowest magnification (ex. 4x) into the
optical path. Focus the microscope
with the coaxial coarse/fine focusing
knob. (The phase contrast objective is
identified as it is impressed with the Ph
code. See page 26.)
16. Bring the phase annular
diaphragm into the optical path.
Identify the annular diaphragm that has
the same Ph code as marked on the
objective now in the optical path, and
bring it into the optical path. (If the
system condenser turret unit is used,
rotate the turret to bring the module
having the same Ph code as marked on
the objective into the optical path.)
1 7. Center the annular diaphragm.
Open the field aperture diaphragm to
the full.
If BT1 eyepiece tube is used:
Set the turret assembly to B. While
observing through both eyepieces,
rotate the Bertrand lens focusing
screw to focus on the phase plate
image of the objective and the annular
diaphragm image of the condenser.
If BT2 eyepiece is used:
Remove an eyepiece and insert the centering telescope in place. Holding the
knurled part of the centering telelscope, rotate its eyepiece to focus on both
the phase plate image of the objective and the annular diaphragm image of the
condenser.
Figure 2-1 7

Adjust so that the phase plate image is
concentric with the annular diaphragm
image. Note that since any
displacement of the phase plate and
annular images will cause low contrast
to the phase contrast image of the
specimen,the exact coincidence of the
two images is necessary.
• System condenser turret unit
Open the condenser aperture
diaphragm to the full. (The optical
path is blocked if the condenser
aperture diaphragm is narrowed.)
Insert two hexagonal screwdrivers into
the annular diaphragm centering
screws of the module in the optical
path, and rotate the screwdrivers to
center the annular diaphragm.
• ELWD condenser
Unclamp the right-hand and left-hand
turret centering knobs. Rotate the
centering knobs for adjustment.
Through PhL centering, other annular
diaphragms are also centered
automatically.
• SLWD condenser
Bring the PhL annular diaphragm into
the optical path, then rotate the
centering screws for adjustment.
Maintain the field aperture diaphragm
fully opened during observation.
(No centering adjustment is necessary
for the Phi annular diaphragm since
it is fixed.)
Figure 2-19
Figure 2-20
10

18. Perform microscopy with a desired phase contrast objective.
• Use the annular diaphragm (module) that has the same Ph code as marked on
the objective on the optical path. Every time the annular diaphragm (module)
is changed, center the annular diaphragm as described in 1 7 . (As for the
ELWD condenser, once centering is complete with PhL, other annular
diaphragms are also centered automatically.)
• Adjust the field aperture diaphragm until it is circumscribed with the viewfield.
(Open it to the full for the SLWD condenser)
• Adjust the brightness with the ND filters or the light intensity control dial.
• If the system condenser turret unit is used, always open the condenser
aperture diaphragm to the full.
• For microscopy at the maximum contrast, bring the GIF filter into the optical
path. Keep the NCB1 1 filter away from the optical path when using GIF filter.
• When replacing the specimen, you can use the refocusing stopper (page 34),
condenser refocusing stopper (page 35), and pillar tilting mechanism (page
36) for enhanced efficiency.
Now, the phase contrast microscopic operation is all completed.
When you observe a specimen in a laboratory dish or other containers, we
recommend the use of an objective that has a correction ring for compensating the
thickness of the dish or container bottom. (See page 26.)

Ill . PHOTOMICROGRAPHIC PROCEDURE
Nikon's 35mm single-lens reflex cameras (such as F-601) and photomicrographic
attachment MICROFLEX Series (option) may be used for photomicrography with the
Diaphot 300.
Mount an SLR camera directly on the microscope front port. Because intermediate
magnification, 2.5x, is applied, the photomicrography magnification is the
"magnification of the objective multiplied by 2.5". The use of a cable release
(option) eliminates the effects of vibration, allowing efficient photomicrography.
Mount a photomicrographic attachment directly on the side port, or on the eyepiece
tube port via the optional eyepiece tube bracket and the trinocular eyepiece tube for
the upright type microscope (option).
The following shows the photomicrographic procedures using a 35 mm SLR
camera.
Before starting photomicrography, make sure that:
♦ The microscope is ready for normal microscopy. (See page 4.)
♦ The camera is loaded with film. (Refer to the instruction manual that comes with
your camera.)
♦ For the photomicrographic procedures using the photomicrographic attachment,
refer to its manual. Rotate the optical path changeover dial to align the optical
path to the port on which the photomicrographic attachment is mounted.
♦ Regarding the procedure for mounting a camera or photomicrographic device, see
Section V . "ASSEMBLY", from page 42.
♦ For detailed operations with individual components, see Section IV .
"OPERATIONS ON COMPONENTS ", from page 16.
1. Turn on the camera.
Flip on the main switch of the camera.
Set the camera for the appropriate film
sensitivity, exposure mode, and
photometry mode. (Refer to the
instructions supplied with the camera.)
Figure 3-1
2. Adjust lamp voltage to PHOTO.
Set the light intensity control dial on the
left side of the base to PHOTO, (page 1 7)
(If the light intensity control dial on the
power supply unit is used, set near "9".)
Figure 3-2

3. S elect the optical path for
photomicrography.
Set the turret assembly to 0. (This
applies to the eyepiece tube BT1 only.)
Set the optical path changeover dial on
the right side of the base to B.
4. S et the filters.
• Daylight type color film: Bring the
NCB1 1 filter into the optical path.
• Monochrome film or tungsten type
color film: Bring the NCB1 1 filter out
of the optical path.
Figure 3-4
5. C ompose the frame.
Turn down the reticle in/out lever to the
right to bring the photomask into the
optical path.
Move the stage to bring the desired
object inside the photomask, (page 39)
6. A djust the focus.
While observing through the eyepieces,
focus on your specimen . (page 40)
13

7. A djust the diaphragm.
Narrow the field aperture diaphragm
until it is slightly wider than the area to
be photomicrographed.
Adjust contrast, depth of focus and
resolution with the condenser aperture
diaphragm. (This diaphragm should
usually be set to a range of 70 to 80%
of the objective's numerical aperture.
See page 22 . )
Figure 3-7
8. C h e ck the exposure time.
Check the exposure time displayed on
the camera. Adjust the brightness so
that the exposure time is longer than
1 /8 of a second.
• Color film: Adjust with the ND filters.
(Keep the NCB1 1 filter in the optical
path.)
• Monochrome film: Adjust with the ND
filters or by varying the lamp voltage.
Figure 3-8
9. P r event external light from
coming in.
Fit the finder cap (option) to the camera
finder. (Keep the finder cap on the
finder except when the exposure time
should be checked through the finder.)
Set the turret assembly to C. (This
applies to the eyepiece tube BT1 ,
especially for long-time exposure.)
Figure 3-9
14