M568 E 11.8.NF.

1 (1/2)

*M568EN01*

Upright Microscope

/
Instruction Manual

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 Introduction

Introduction

Thank you for purchasing a Nikon product.

This instruction manual is written for users of the Nikon ECLIPSE Ci-S and Ci-L microscopes. To ensure correct usage,
read this manual carefully before operating this product.

● No part of this manual may be reproduced or transmitted in any form without prior written permission from Nikon.
● The contents of this manual are subject to change without notice.
● Appearance of the product shown in this manual may be different from that of your product.
● Although every effort has been made to ensure the accuracy of this manual, errors or inconsistencies may remain. If
you note any points that are unclear or incorrect, please contact your nearest Nikon representative.
● Some of the equipment described in this manual may not be included in the set you have purchased.
● If you intend to use any other equipment with this product, read the manual for that equipment too.
● If this equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may
be impaired.
● Training: This product can be used without special training, provided that this manual is read thoroughly before use. Kindly
contact your nearest Nikon representative if you have any questions, find any errors, or wish to provide us with your
opinion.

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 Introduction

Contents of the Manual

The manual for ECLIPSE Ci-S/Ci-L consists of the following contents. There is also a "Bright-field Microscopy Quick
Guide", provided as a spearate document.

◆ This manual: Instructions

Safety Precautions
Microscopy Procedures
Bright-field Microscopy
Phase Contrast Microscopy
Simple Polarizing Microscopy
Sensitive Tint Plate Microscopy
Epi-fluorescence Microscopy
Individual Operations
Assembly
Troubleshooting
Maintenance and Storage
Specifications and Safety Standards

◆ Bright-field Microscopy Quick Guide

Symbols Used in This Manual

The following symbols are used in this manual.

◆ Symbols for Safety

WARNING
Highlights important information that should be noted for safety. Read "Safety Precautions" for details.
CAUTION

◆ Other Symbols

Indicates information you should note or comply with to prevent defects or malfunction of this product.

Indicates information you should be aware of in using this product, as well as other useful information.

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 Table of Chapters

Table of Chapters (See next page for the detailed contents.)

Introduction Safety Precautions/

Contents of the Manual Notes on Handling the Product
Symbols Used in This Manual
Chapter 1-1
Microscopy Procedures
Bright-field Microscopy

Chapter 1-2
Microscopy Procedures
Phase Contrast Microscopy

Chapter 1-3
Microscopy Procedures
Simple Polarizing Microscopy

Chapter 1-4
Microscopy Procedures
Sensitive Tint Plate Microscopy

Chapter 1-5
Microscopy Procedures
Epi-fluorescence Microscopy

Chapter 2
Individual Operations

Chapter 3
Assembly

Chapter 4
Troubleshooting

Chapter 5
Maintenance and Storage

Chapter 6
Specifications and Safety Standards

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 Table of Contents

Table of Contents

Introduction ......................................................................................................................... i
Contents of the Manual ....................................................................................................ii
Symbols Used in This Manual ..........................................................................................ii
Table of Chapters.............................................................................................................. iii
Safety Precautions............................................................................................................ vi
WARNING and CAUTION Symbols used in This Manual...............................................vi
Meaning of Symbols Used on the Product ......................................................................vi
WARNING ................................................................................................................ vii
CAUTION ..................................................................................................................ix
Notes on Handling the Product.......................................................................................... x

Microscopy Procedures .......................................................................... 1

1 Bright-field Microscopy..............................................................................................1
1.1 System Configuration and Controls ................................................................... 1
1.2 Bright-Field Microscopy Procedure .................................................................... 2
2 Phase Contrast Microscopy ......................................................................................9
2.1 System Components and Controls .................................................................... 9
2.2 Phase Contrast Microscopy Procedure ........................................................... 10
3 Simple Polarizing Microscopy .................................................................................15
3.1 System Configuration and Controls ................................................................. 15
3.2 Simple Polarizing Microscopy Procedure ........................................................ 16
4 Sensitive Tint Plate Microscopy ..............................................................................21
4.1 System Configuration and Controls ................................................................. 21
4.2 Sensitive Tint Plate Microscopy Procedure...................................................... 22
5 Epi-fluorescence Microscopy ..................................................................................27
5.1 System Configuration and Controls ................................................................. 27
5.2 Epi-fluorescence Microscopy Procedure ......................................................... 28

Individual Operations ............................................................................ 33

1 Adjusting the Brightness of a Diascopic Image.......................................................33
1.1 Adjustment with the Dia-illumination Brightness Control Knob........................ 33
1.2 Adjustment with the ND Filter (for Ci-S)........................................................... 33
1.3 Removing an NCB filter for a brighter image (Ci-S)......................................... 34
2 Focusing on the Specimen (Vertical Stage Movement)...........................................35
2.1 Focus Knob Rotation and Stage Movement .................................................... 36
2.2 Number of Focus Knob Turns and Distance of Stage Travel........................... 36
2.3 Adjusting the Rotating Torque of the Coarse Focus Knob............................... 37
2.4 Refocusing ....................................................................................................... 37
2.5 Position Exchange of the Fine Focus Knob ..................................................... 38
3 Bringing the specimen into the optical path (Horizontal Stage Movement) .............39
3.1 Knob Rotation and Stage Movement ............................................................... 39
3.2 Adjusting the Knob Heights.............................................................................. 39
3.3 Adjusting the Knob Rotation Torque................................................................. 39
4 Adjusting the Diopter...............................................................................................40
5 Adjusting aperture diaphragm .................................................................................41
6 Adjusting the Aperture Diaphragm ..........................................................................42
6.1 Adjusting the Aperture Diaphragm Using the Condenser Scale...................... 42
6.2 Adjusting the Aperture Diaphragm Using the Centering Telescope................. 42
7 Selecting a Condenser............................................................................................43
8 Adjusting the field diaphragm..................................................................................43

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 Table of Contents

9 Switching the Optical Path of the Tube ...................................................................44

9.1 Light Distribution............................................................................................... 44
9.2 Disabling the Clicking of the Optical Path Switching ....................................... 44
10 Adjusting the Binocular Section ..............................................................................45
10.1 Adjusting with the Binocular Section of the Tube............................................. 45
10.2 Using the Eyelevel Riser .................................................................................. 45
11 Using Stage at Lowered Position (Spacer for Nosepiece) ......................................45
12 Oil Immersion ..........................................................................................................46
13 Water Immersion .....................................................................................................47
14 Tips for Phase Contrast Microscopy .......................................................................47
15 Tips for Epi-fluorescence Microscopy .....................................................................50
15.1 Switching Excitation Methods .......................................................................... 51
15.2 Selecting Filters................................................................................................ 52
16 Capturing Images....................................................................................................54
16.1 Photomicroscopy.............................................................................................. 54
16.2 Tips on Microscope Settings for Photomicroscopy .......................................... 55

Assembly................................................................................................ 57
1 ECLIPSE Ci-S/Ci-L System Configuration ..............................................................57
2 Assembly for Bright-field Microscopy ......................................................................58
3 Assembly for Phase Contrast Microscopy ..............................................................62
4 Assembly for the Simple Polarizing Microscopy .....................................................62
5 Assembly for Sensitive Polarization Microscopy.....................................................63
6 Assembly for Epi-fluorescence Microscopy ............................................................64
7 Attaching a Camera ................................................................................................66

Troubleshooting..................................................................................... 67
1 Optical System and Operation ................................................................................67
1.1 General............................................................................................................. 67
1.2 Epi-fluorescence Microscopy ........................................................................... 71
1.3 Phase Contrast Microscopy ............................................................................. 72
2 Electrical requirements............................................................................................73
2.1 General............................................................................................................. 73
2.2 Epi-fluorescence Microscopy ........................................................................... 73

Maintenance and Storage ..................................................................... 75

1 Replacing the Lamp (for Ci-S).................................................................................75
2 Cleaning ..................................................................................................................76
2.1 Cleaning Lenses............................................................................................... 76
2.2 Cleaning Parts Other than Lenses ................................................................... 76
2.3 Cleaning the Immersion Oil.............................................................................. 77
2.4 Decontaminating the Product........................................................................... 77
3 Storage....................................................................................................................77
4 Regular Inspections (Charged) ...............................................................................77

Specifications and Safety Standards................................................... 79

1 Microscopy (Operation Principle) ............................................................................79
2 Performance Properties ..........................................................................................79
3 Physical Properties .................................................................................................81

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 Safety Precautions

Safety Precautions

To ensure correct and safe operation, read this manual before using this product.

WARNING and CAUTION Symbols used in This Manual

Safety Precautions/Notes on Handling the Product

Although this product is designed and manufactured to be completely safe during use, incorrect usage or failure to follow
the safety instructions provided may cause personal injury or property damage. To ensure correct usage, read this manual
carefully before using this product. Do not discard this manual and keep it handy for easy reference.

Safety instructions in this manual are marked with the following symbols to indicate their importance. For your safety,
always follow the instructions marked with these symbols.

Symbol Description

WARNING Disregarding instructions marked with this symbol may lead to serious injury or death.

CAUTION Disregarding instructions marked with this symbol may lead to injury or property damage.

Meaning of Symbols Used on the Product

When appearing on this product, the symbols below indicate the need for caution at all times during use. Read the
relevant instructions in this manual before attempting to use or adjust any part to which the symbol has been affixed.

Biohazard

This symbol is affixed to the front of the stand of this product, to call your attention to the following:

● WARNING: Using this product may constitute a biohazard risk if a sample comes into contact with this
product.

● To avoid biohazard contamination, do not touch the contaminated portion with bare hands.

● Decontaminate the contaminated part according to the standard procedure specified for your
laboratory.

Precautions against heat

This symbol is affixed to the part near the lamphouse of the ECLIPSE Ci-S to call your attention to the
following:

● During and immediately after a period of illumination, the lamp and surrounding areas (including the
lamphouse) are very hot.

● Risk of burns. Do not touch the lamp or surrounding areas during or immediately after a period of
illumination.

● Make sure the lamp and surrounding areas have cooled sufficiently before attempting to replace the
lamp.

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 Safety Precautions

WARNING
1 Do not disassemble. ● Always attach the lamphouse cover when using
Disassembling this product may result in electric this product.
shock or malfunction. Malfunction and damage ● Make sure the lamp and surrounding areas

Safety Precautions/Notes on Handling the Product

due to disassembling or modification are have cooled sufficiently before attempting to
unwarranted. replace the lamp (about thirty minutes).
Do not disassemble parts other than those ● To avoid the risk of fire, do not place fabric,
described in this manual. If you experience paper or highly flammable volatile materials
problems with this product, contact your nearest such as gasoline, petroleum benzine, paint
Nikon representative. thinner, or alcohol near the lamphouse while
the lamp is lit or for a period of approximately
2 Read the instruction manuals carefully. thirty minutes after the lamp is turned off.
To ensure safety, thoroughly read this manual and
5 Hazards of mercury lamps
the manuals for other equipment to be used with
(when using the CI-FL Epi-fluorescence
this product. Particularly, all warnings and
attachment)
cautions given at the beginning of each manual
The light source used with the epi-fluorescence
must be observed.
attachment (HG Precentered Fiber Illuminator)
Safety is a top design priority for Nikon products.
requires special care during handling because of
Safety is ensured as long as the user observes all
its characteristics. For safe and correct use of this
of the warnings and cautions given in the
system, carefully read the warnings below. Keep
manuals, and uses the system only for its
in mind all potential hazards. Additionally, carefully
intended purpose. However, failure to heed the
read the manual for the illuminator and the manual
warnings and cautions given in the manuals,
from the lamp manufacturer (if provided), then
subjecting the system to shock or impact, or
follow the instructions given therein. Failure to
attempting to disassemble the system may result
heed the warnings and cautions given in the
in unexpected accidents and injury.
manuals, subjecting the system to shock or
Product with CI-FL epi-fluorescence
impact, or attempting to disassemble the system
attachment: may result in unexpected accidents and injury.
The light source used for Epi-fluorescence
● Ultraviolet light
microscopy (HG Precentered Fiber Illuminator)
When lit, mercury lamps radiate ultraviolet light
requires special care during handling because of
that can damage the eyes and skin. Direct
its characteristics. Be sure to refer to the manual viewing of the light may result in blindness.
for the light source being used. When changing filter cubes, always turn off the
3 Notes on the power cord light source of the Epi-fluorescence
Be sure to use the specified power cord. Use of attachment. Leaving the lamp turned on during
other power cords may result in malfunction or filter cube replacement may result in ultraviolet
fire. This product is classified as having Class I exposure.
protection against electric shock. Make sure this ● High-pressure gas
product is connected to an appropriate protective The lamps contain sealed gas under very high
earth terminal. pressure. And the pressure increases when the
lamp is on. If the lamp is scratched, fouled,
Refer to Chapter 6, “2 Performance Properties” for
subjected to high external pressure or physical
the specified power cords.
impact, or used beyond its service life, the
● To prevent electric shock, always turn off the sealed gas may leak or the lamp may burst,
power switch (press to the “O” position) for the resulting in gas inhalation, injury from glass, or
microscope before connecting or disconnecting other accidents.
the power cord. ● Heat
4 Heat from the illuminator When the lamp is lit, the lamp and surroundings
(when using ECLIPSE Ci-S) will become extremely hot. Do not touch the
During and immediately after a period of illumination, lamp with bare hands or place flammable
the lamp and surrounding areas (including the materials near the lamp. Failure to comply may
result in burns or fire.
lamphouse) are very hot.
● Designated lamp
● Do not touch the lamp or surrounding areas
Be sure to use the designated lamp. Using
during or immediately after a period of
other types of lamps may result in accidents,
illumination. There is a risk of burn if you touch
including bursting of the lamp.
the hot area.

vii

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 Safety Precautions

WARNING
6 Hazardous sample handling
This product is intended primarily for microscopic
observations and image capture of cells and
Safety Precautions/Notes on Handling the Product

tissue set on glass slides.

Check to determine whether a sample is
hazardous before handling. If sample is
hazardous, handle it according to the standard
procedure specified for your laboratory. If the
sample is potentially infectious, wear rubber
gloves and avoid directly touching samples. If
such a sample is spilled onto this product, the
portion must be decontaminated in a safety
manner. Consult your safety supervisor or safety
standard of your facility.

viii

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 Safety Precautions

CAUTION
1 Power shutdown 6 Cautions on assembling, installing, and
To prevent electric shock and/or malfunction, carrying the product
always turn off the power switch(es) for this ● Take care to avoid pinching your fingers or

Safety Precautions/Notes on Handling the Product

product and the peripheral devices (press to the hands during product assembly and
“O” position) and unplug the power cord from the installation.
wall outlet before assembling this product, ● Scratches or fouling optical components (such
connecting or disconnecting cables, replacing as lens and filters) with fingerprints, etc. will
lamps, or cleaning this microscope and the degrade microscope images. Be careful to
objective. avoid scratches or direct contact with the lens
2 Lamp replacement precautions and filters when assembling.
(when using ECLIPSE Ci-S) ● The main body weighs approximately 10 kg.
● To avoid burns, wait at least 30 minutes after When moving it, hold the main body by the grip
the lamp is turned off to give it sufficient time to on the back and the recessed portion at the
cool. To avoid electric shock or malfunctions, front bottom side of this product.
never attempt to replace the lamp without first ● Remove all attachments (if mounted) from the
turning off the power switches for this product microscope before moving the microscope.
and the peripheral devices (press to the “O”
● Do not place this product in a locker or cabinet.
position) and unplugging the power cord from
the wall outlet. 7 Cautions on use, transportation, and
● Make sure the lamphouse cover is securely storage
fitted to the lamphouse after lamp replacement. This product must be operated, transported, or
Never turn on the lamp while the lamphouse stored in accordance with the following conditions.
cover is open. Installing this product in a hot, humid location may
● Do not break used lamps. It should be disposed result in the formation of mold or condensation on
of as industrial waste, in accordance with local lenses, impairing performance or causing
regulations and rules. malfunctions.

3 Designated lamp ● Operating conditions:

(when using ECLIPSE Ci-S) temperature: 0 to +40°C,
The product's built-in power source is used for humidity: 60% RH max. (no condensation)
lighting the halogen lamp that is a light source for ● Transporting/storage conditions:
dia-illumination. Halogen lamps of up to 6V-30W temperature: -20 to +60°C,
can be lit. Always use the specified halogen lamp. humidity: 90% RH max. (no condensation)
Using an unspecified lamp may cause
8 Remove any covers from the product before
malfunctions.
switching on.
Designated lamp: 6V-30W (PHILIPS 5761)
Do not use this product while it is covered with a
4 Prevent contact with water. piece of cloth, etc., This will result in an abnormal
Never allow water to come into contact with this heating or a fire hazard. Do not cover this product
product, and avoid using this product in with a piece of cloth or similar while in use. The
circumstances where it may be splashed with system temperature will rise, resulting in a
water. Splashing water onto this product may malfunction.
cause a short circuit, resulting in malfunction or 9 Cautions on sustained observations
abnormal heating. If water is splashed onto this
To relieve fatigue resulting from long observation
product, immediately turn off the power switches
sessions, limit continuous observations to one
for this product and the peripheral devices (press
hour. Take at least 10 to 15 minutes breaks
to the “O” position) and remove the power cord
between observation sessions. Adjust the layout
from the receptacle. Then wipe off moisture with a
of other equipment used and the height of your
piece of dry cloth or something similar. If water
chair.
enters, stop the use of the product and contact
your nearest Nikon representative. 10 Cautions on the disposal of the product
5 Do not place any object on top of the To avoid biohazard risks, dispose of this product
product. as contaminated equipment in accordance with
the standard procedure specified for your facility.
Do not place any object on top of this product.

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 Safety Precautions

Notes on Handling the Product

1 Handle with care ● Select a location with minimal dust.

This product is a precision optical instrument and ● To avoid splashes, do not use this product near
requires gentle handling. Avoid subjecting it to water.
Safety Precautions/Notes on Handling the Product

sudden impacts and shocks. ● Make sure the ambient temperature is 0 to +40°C
Even relatively minor impacts are capable of affecting and humidity is 60% or less. When transporting or
the precision of the objective. storing this product, the ambient temperature
must be -20 to +60°C, with the humidity at 90%
2 Weak electromagnetic waves RH max (with no condensation). Installing this
This product emits weak electromagnetic waves. So product in a hot, humid location may result in the
as to avoid degrading the performance of precision formation of mold or condensation on lenses,
electronic devices, do not install this product near impairing performance or causing malfunctions.
such devices. If TV or radio reception is affected, ● Do not place this product in a locker or cabinet.
move the TV or radio further from this product.
6 Handling of focus knob
3 Dirt on the lens ● Never turn the focus knobs on the right and left
Scratches or fouling optical components (such as sides of the microscope in opposite directions at
lens and filters) with fingerprints, etc. will degrade the same time. Doing so may damage this
microscope images. product.
If these parts become dirty, clean them as described ● Turning the coarse focus knob past its farthest
in Chapter 5, “2.1 Lens Cleaning”. point will damage this product. Never use undue
force when turning the knob.
4 Dirt on the lamp (ECLIPSE Ci-S)
Never touch the lamp with bare hands. Dirt or 7 Protect the ports from dust and extraneous
fingerprints on the lamp will result in uneven light (when the trinocular tube or the
illumination and reduce the service life of the lamp. ergonomic tube is attached).
Always wear gloves when handling lamps.
To keep out extraneous light and dust, always attach
5 Installation location the supplied cap to any port not currently in use.
This product is a precision instrument. Usage or 8 Handling of filters (when using the
storage of this product in an inappropriate epi-fluorescence attachment)
environment may result in malfunction or a
● Excitation filters inside a filter cube are exposed
degradation in precision. Consider the following
to strong light and degrade over time. Replace
factors when selecting an installation location:
them after the appropriate number of hours.
● Select a location free of vibration. Install this
● Filter characteristics may alter if the filter is
product on a level surface.
exposed to high humidity. To prevent changes or
● Install this product at least 10 cm away from walls. degradation of filter characteristics, avoid using or
● Choose a location less exposed to hazards in the storing the filters under conditions of high
event of collisions, earthquakes, or other potential humidity or high temperature. Avoid subjecting
disasters. To keep this product from falling, use a filters to rapid temperature changes. When a filter
strong rope or other means if necessary to secure is not in use, store in a desiccator or hermetically
it to the working desk or other heavy, stable item. sealed container with a drying agent.
● Select a layout that allows easy removal of the ● Especially the filters in the nine types of filter
power cord from the product's AC inlet in the event cubes listed below offer sharp, high-resolution
of an emergency. waveform characteristics superior to normal
● Do not use a desk mat or similar. filters. However, due to their sophisticated
coatings, they must be handled with special care.
● Avoid locations exposed to direct sunlight,
Take care to avoid abrasion from cleaning. Follow
locations immediately under room lights, and other
the description in “2.1 Lens Cleaning” in Chapter
bright locations.
5.
● Light from room lights just above this product may
enter the objective as extraneous light. If possible, Single-band filter cubes:
switch off the room lights directly above this DAPI, FITC, TxRed, GFP
product when making observations.
Multi-band filter cubes:
F-R, F-T, D-F, D-F-R, D-F-T

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 1 Microscopy Procedures

1 Bright-field Microscopy

1.1 System Configuration and Controls

This section explains an example system configuration and the controls required for bright-field microscopy using the
ECLIPSE Ci-S/Ci-L.
Names of components are denoted in the following manner: [Eyepiece]. Chapter 1-1

Diopter adjustment ring

Microscopy Procedures
[Eyepiece] Optical path switching lever

[Tube]

[Nosepiece] Condenser focus knob

Bright-field Microscopy
Coarse focus clamp ring

[Objective] Coarse focus knob

Fine focus knob

[Stage]
Y knob
Aperture diaphragm lever
TO RQ

Power switch
UE

[Condenser]
P
CL AM

8
ND4 ND
OUT

ND filter IN/OUT switch

IN
(equipped with Ci-S only)
[Main body] X knob
(The figure shows
ECLIPSE Ci-S.) Capture button

Field diaphragm dial

Moving claw of the

Grip specimen holder

Condenser focus knob

Condenser centering
NIKON

screw
Input voltage label
CORP
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MODEL
100–24 ECLIPSE
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conditi ion is Part 15
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Coarse focus torque

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adjustment knob
Canada
.

Coarse focus knob

90
0
80
10
70

20

Fine focus knob

60

30
50

AC inlet
40

DS C

Dia-illumination
DSC connector brightness control knob

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 Chapter 1 Microscopy Procedures

1.2 Bright-Field Microscopy Procedure

1. Turn on the power. 7

2. Lower condenser slightly from
uppermost position.
14
3. Fully open field and aperture
diaphragms.
Chapter 1-1 8
4. Bring the 10x objective into the optical
path.
Microscopy Procedures

5. Bring specimen into optical path.

6. Focus on specimen.
7. Adjust diopter. 4 10
2 9
8. Adjust interpupillary.
9. Focus and center condenser. 6 12
3 11
10. Bring the desired objective into the
optical path. 9 1 15

TO RQ
UE
Bright-field Microscopy

P
11. Adjust the aperture diaphragm.

CL AM
ND4 ND8
OUT

12. Focus on specimen. IN

13. Circumscribe field diaphragm to field

of view. 5
14. View specimen. 3 13
15. Turn off the power.

Preparation for microscopy

1 Turn on the power.

Press the switch to the “|” position to turn on the power to
the microscope (the power LED on the front of the main ON
body will light up to indicate that dia-illumination is turned
ON).

Power on

POWER

LED on

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 Chapter 1 Microscopy Procedures

2 Lower the condenser slightly from the uppermost position.

Turn the condenser focus knob until the condenser is
positioned at the upper limit (where it clicks to a stop), and
then lower it a little.

TO RQ
UE

P
CL AM
Chapter 1-1
8
ND4 ND
OUT

Microscopy Procedures
IN

Lower the condenser from upper limit

3 Fully open the field diaphragm and aperture

diaphragm.
Turn the field diaphragm dial and the aperture diaphragm
lever clockwise to open them completely.

TO RQ
UE

Bright-field Microscopy
P
CL AM
8
ND4 ND
OUT

IN

Fully open field and aperture diaphragms.

4 Bring the 10x objective into the optical path.

Turn the nosepiece to bring the 10x objective into the
optical path.

Turn the nosepiece until it clicks.

Bring the 10x objective into the optical path.

5 Place a specimen on the stage, and move the

stage to bring the target into view.
(1) Open the claw of the specimen holder's moving part
and place the specimen onto the stage, gently
stowing the claw back to fix the specimen.

Setting the specimen

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 Chapter 1 Microscopy Procedures

(2) Rotate the stage knob to move the stage and bring
the target into the optical path.
(So that the sample sealed under the cover glass
will be lighted.)
0.8 0.6 0.4 0.2

Chapter 1-1
Microscopy Procedures

POWER

Bringing the Target into the Optical Path

6 Focus on the specimen. (→See Chapter 2 “2 Focusing on the Specimen (Vertical Stage Movement)” for
details)

(1) When using the trinocular tube or ergonomic tube,

push in the optical path switching lever to distribute
100% light to the binocular section.
Bright-field Microscopy

(2) Look into the eyepiece and turn the coarse focus
knob away to raise the stage to the upper limit. From
there, focus on the specimen by lowering the stage.

Switching the Optical Path 100% to the Binocular

Part

(3) When the focus was roughly adjusted using coarse

focus knob, turn the fine focus knob to accurately
adjust the focus.

0.8 0.6 0.4 0.2

POWER

Focusing on the Specimen

CLA

(4) Adjust the brightness of the field of view by turning

the dia-illumination brightness control knob.
80
TO R

90
QU
E
0
10
20

30

PO WER

Brightness Adjustment

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 Chapter 1 Microscopy Procedures

Notes on controlling the focus knobs

Avoid the following action, which can cause equipment
malfunction.
0.8 0.6 0.4 0.2

• Rotating the right and left focus knobs in opposite directions.

• Rotating the coarse focus knob past the limit.

POWER

Chapter 1-1

Microscopy Procedures
Don't rotate the knobs in opposite directions!

7 Adjust the diopter. (→See Chapter 2, “4 Adjusting the

Diopter” for details)

(1) Turn the diopter adjustment ring on the right and left
eyepieces to align the end face of the diopter Line
adjustment ring with the line. (This is the diopter
adjustment reference position.)

Bright-field Microscopy
(2) Focus on the specimen using the 40x objective.
(3) Bring the 10x (or 4x) objective into the optical path. Reference position for diopter adjustment

(4) Look into the right eyepiece with your right eye and
the left eyepiece with your left eye. Turn the diopter
adjustment ring of each eyepiece to focus on the
specimen. At this point no focus knobs are used.

(5) Repeat Steps (2) through (4) to make sure the focus
Adjusting the Diopter
has been adjusted properly.

8 Adjust the interpupillary distance.

Look into both eyepieces and rotate the binocular part to
adjust the binocular part's opening until the fields of view
for the right and left eyes coincide.

For easy adjustment, look into the eyepiece as if

you were looking at a distant object.

Adjusting Interpupillary Distance

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 Chapter 1 Microscopy Procedures

9 Focus and center the condenser. (See Chapter 2, “5 Focusing and Centering the Condenser” for details)

(1) Look into the eyepiece with the field diaphragm

stopped down to the minimum. Focus on the field
diaphragm image using the condenser focus knob, 0.8 0.6 0.4 0.2

then adjust the condenser centering screws to

center the diaphragm image within the field of view.

Chapter 1-1 (2) Bring the 40x objective into the optical path to check POWER

the focus and centering of the field diaphragm

Microscopy Procedures

image. Make adjustments in the same way as step Field diaphragm

(1) as necessary. image

Eyepiece field
of view

(3) Turn the field diaphragm dial and adjust the field
Bright-field Microscopy

diaphragm image so that its size is almost the same

as the field of view.

Field diaphragm
image

Eyepiece field
of view
Adjusting aperture diaphragm

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 Chapter 1 Microscopy Procedures

Microscopy operation

10 Select the desired objective.

Turn the nosepiece to bring the desired objective into the
optical path.

Swing-out condenser 1-100x

Mounting a 1-100x swing-out condenser on the Chapter 1-1
Ci-L main body when using the 1x objective may

Microscopy Procedures
result in uneven illumination around the field of
view.

Bring an arbitrary objective into the optical path.

11 Adjust the aperture diaphragm. (→See Chapter 2,

“6 Adjusting the Aperture Diaphragm” for details)
Aperture

Bright-field Microscopy
Turn the aperture diaphragm lever on the condenser to diaphragm image
adjust the aperture diaphragm so that it is set to 70 to 80% Objective 70 to 80
of the numerical aperture of the objective used. pupil

100
Be sure to adjust the aperture diaphragm each
time you change the objective.
Right size of the aperture diaphragm
(You can see the aperture diaphragm image with
the centering telescope.)

12 Focus on the specimen.

(1) Look into the eyepiece, and adjust the brightness of

the field of view by turning the dia-illumination
brightness control knob. You can also adjust the
brightness with an ND filter for Ci-S.
(2) Rotate the stage knob to move the stage and bring
the target into the optical path.
(3) If the specimen is not in focus, turn the focus knob to
focus on it.

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 Chapter 1 Microscopy Procedures

13 Adjust the field diaphragm

Turn the field diaphragm dial to adjust the field diaphragm
so that it almost circumscribes the field of view.

Size of the field diaphragm Field diaphragm

Normally, adjust the field diaphragm so that it dial
almost circumscribes the field of view. Opening
the field diaphragm too much results in stray light
entering the field of view, generating flare and
Chapter 1-1 Field Diaphragm Field of view
reducing the image contrast. In addition, the
specimen will become decolorized over a wider
Microscopy Procedures

area.

Field diaphragm's adjustment timing

Be sure to adjust the field diaphragm each time
you change the objective.
Circumscribe around the field of view

Adjusting the field diaphragm

Bright-field Microscopy

14 View the specimen.

Rotate the stage knob to move the target. If the target is
not in focus, use the focus knob to adjust the focus.

15 Turn off the power.

Turn off the power switch (press to the “O” position) for the
microscope. (The power LED on the front of the main
body will turn off.) 8
ND4 ND
OUT

IN OFF

Power off

POWER

Power LED off

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 Chapter 1 Microscopy Procedures

2 Phase Contrast Microscopy

2.1 System Components and Controls

This section explains an example system configuration and the controls required for phase contrast microscopy using the
ECLIPSE Ci-S/Ci-L.
Names of components are denoted in the following manner: [Phase contrast condenser].

Diopter adjustment ring

[Eyepiece]

Optical path Chapter 1-2

switching lever

Microscopy Procedures
[Tube]

[Nosepiece] Condenser focus knob

Coarse focus clamp ring

[Objective]
Coarse focus knob

Fine focus knob

[Stage]

Phase Contrast Microscopy

P
H
A
S 0.9 JAP
E 0 AN
CD
O RY
N
T
R
A
S
T

Y knob
A

Aperture diaphragm lever

TO RQ
UE

[Phase turret condenser] Power switch

P
CL AM

8
ND4 ND

Condenser turret OUT

ND filter IN/OUT switch

IN
(equipped with Ci-S only)
[Main body] X knob
(The figure shows
ECLIPSE Ci-S.) Capture button

Field diaphragm dial

[Centering telescope]
Moving claw of the
[GIF filter] specimen holder
(green interference filter)
Condenser focus knob

Grip Condenser centering screw

Annular diaphragm centering
knob
Annular diaphragm
NIKON centering knob
Input voltage
CORP
TOKY ORAT
O, JAPAN ION
MODEL
100–24 ECLIPSE
0V~ Ci-S
0.9A

Coarse focus torque

50/60H
z 9400
This
device MADE 01
Rules. complie IN CHINA
Operat s with
conditi ion is Part 15

label
ons: subject of the
(1) this to the FCC
4N75 device followin
INSPE and (2) may g two
this device not cause
EQUIPCTION receive
must harmfu
MENT d, l interfer
undesi includin g accept
any interfer ence,
red operati interfer

adjustment knob
ence
This on. that may ence
Class cause
Canadi A digital
an appara
Cet appareICES-0 03. tus complie
confirm il numéri s with
e à la que de
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NMB-0 A est
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.

Coarse focus knob

80
90
0

Fine focus knob

Lamphouse
10
70

20
60

30
50

40

AC inlet DS C

Dia-illumination
DSC connector brightness control knob

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 Chapter 1 Microscopy Procedures

2.2 Phase Contrast Microscopy Procedure

In this procedure, only step titles are shown for operations that are the same as those of bright-field microscopy. See “2.1
Bright-field Microscopy” for details.

1. Turn on the power.

8
2. Lower condenser slightly from
uppermost position.
3. Fully open field and aperture 17
diaphragms.
4. Move the turret to the [A: empty] 9
position.

Chapter 1-2 5. Bring the 10x Ph objective into the

optical path.
Microscopy Procedures

6. Bring specimen into optical path.

5 13
7. Focus on specimen. 2 10
8. Adjust diopter.
7 15
9. Adjust interpupillary. 4 11 14

P
H
A
S 0.9 JAP
E 0 AN
CD
O RY
N
T
R
A
S
T
A
10. Focus and center condenser. 1 18
3

TO RQ
UE
11. Bring the Ph annular diaphragm [Ph1]

P
CL AM
position into the optical path. 12 OUT
ND4 ND8

12. Center the Ph annular diaphragm.

IN
Phase Contrast Microscopy

10
13. Bring the desired Ph objective into the
6
optical path.
3 16
14. Match the Ph codes of annular
diaphragm and the objective.
15. Focus on specimen.
16. Circumscribe field diaphragm to field
of view.
17. View specimen.
18. Turn off the power.

Preparation for microscopy

1 Turn on the power.

(The power LED on the front of the main body will light
up.)

2 Lower the condenser slightly from the uppermost position.

3 Fully open the field diaphragm and aperture

diaphragm.

10

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 Chapter 1 Microscopy Procedures

4 Move the condenser turret to the [A: empty]

position.
Turn the condenser turret until the [A: empty] symbol
comes to the front, and align the hole with the optical path.

Aperture diaphragm of the condenser

Positioning the condenser turret at positions other
than [A: empty] deviates the aperture diaphragm

P
H
A
S 0.9 JAP
E 0 AN
A: empty

CD
O RY
N
T
R
A
S
T
A
from the optical path.

TO RQ
UE

P
CL AM
Move the condenser turret to the [A: empty] Chapter 1-2
position.

Microscopy Procedures
5 Bring the 10x Ph objective (Ph1) into the optical
path.

6 Place a specimen on the stage, and move the

stage to bring the target into view.

7 Focus on the specimen. (→See Chapter 2 “2 Focusing on the Specimen” for details)

Phase Contrast Microscopy

8 Adjust the diopter. (→See Chapter 2, “4 Adjusting the Diopter” for details)

9 Adjust the interpupillary distance.

10 Focus and center the condenser. (See Chapter 2, “5 Focusing and Centering the Condenser” for details)

11

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 Chapter 1 Microscopy Procedures

Microscopy operation (→See also: Chapter 2, Section 14 “Tips for Phase Contrast Microscopy”)

11 Bring the Ph annular diaphragm (Ph1) in the condenser turret into the optical path.
Turn the condenser turret until the [Ph1] symbol comes to
the front.

GIF filter Ph1

The GIF filter (green interference filter) improves Condenser

the contrast when placed in the optical path. The turret
filter should be installed on the field lens, or

P
H
A
S 0.9 JAP
E 0 AN
placed inside or on top of the filter cassette holder.

CD
O RY
N
T
R
A
S
T
A
Note, however, that it may cause ghosting when
mounted inside the filter cassette holder.

TO RQ
UE
Chapter 1-2

P
CL AM
Microscopy Procedures

GIF filter (green interference filter)

for improving the contrast of a phase image

Bring the Ph annular diaphragm into the optical

path.

12 Center the Ph annular diaphragm.

To optimize the phase effect, it is important to properly
Eyepiece Flange
overlap the phase plate of the objective with the Ph
Phase Contrast Microscopy

annular diaphragm image in the condenser.

(1) Make sure that the 10x objective (Ph1) has been
placed into the optical path and that the [Ph1]
symbol on the condenser turret is facing the front.
(2) Rotate the stage knob to move the specimen and
Focusing on the Phase Ring
bring a portion where there is no sample under the
cover glass into the optical path.
(3) Remove one eyepiece from the tube, and insert the
centering telescope with adapter into the tube.
(4) Hold the flange of the centering telescope and rotate
the eyepiece to focus on the phase plate of the
objective.
P
H
A
S 0.9 JAP
E 0 AN
CD
O RY
N
T
R

Annular
A
S
T
A

(5) If the phase plate of the objective and the annular diaphragm
TO RQ

diaphragm in the condenser are misaligned, loosen centering knob

UE

the clamp screws of the two annular diaphragm

P
CL AM

Annular diaphragm
centering knobs before rotating the centering knob centering knob clamp
to move the entire turret for annular adjustment. screw
Tighten the clamp screws after the completion of the
adjustment.
(6) Remove the centering telescope and adapter from
the tube, and reattach the eyepiece.
Bad Good

Centering the Ph annular diaphragm

12

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 Chapter 1 Microscopy Procedures

13 Bring an arbitrary Ph objective into the optical

path.
Turn the nosepiece to bring the desired Ph objective into the optical path.
When using an oil immersion type objective, apply immersion oil between the specimen and the objective.
(→Chapter 2 “12 Oil Immersion” for details)

Phase turret condenser

The phase turret condenser is intended to be used dry. DO NOT apply immersion oil between the condenser
tip's ball and the specimen.

14 Adjust the Ph annular diaphragm in the condenser with the Ph objective to be used.
Turn the condenser turret to bring an annular diaphragm
Same Ph code Chapter 1-2
with the same Ph code as the objective into the optical
path.

Microscopy Procedures
P
H
A
S 0.9 JAP
E 0 AN
CD
O RY
N
T
R
A
S
T
A

TO RQ
UE

P
CL AM

Phase Contrast Microscopy

Matching the Ph codes of Ph annular
diaphragm and the Ph objective
Ph code
One of the Ph codes, [Ph1], [Ph2], or [Ph3] is indicated on the Ph objective depending on the size of the phase
plate. (Ph codes have nothing to do with the magnification of the objective.) Always use a Ph objective and Ph
annular diaphragm with the same Ph code. You cannot experience the phase effect if a different combination
of the codes might be used.

Centering of the annular diaphragm and the phase plate

The position of each annular diaphragm in the condenser turret has already been adjusted based on the Ph1
annular diaphragm, but the phase image will differ slightly depending on how the annular diaphragm overlaps
the phase plate. For a stricter observation or capturing of still images, check whether the annular diaphragm
and the phase plate are concentric at each magnification.

15 Focus on the specimen.

(1) Look into the eyepiece, and adjust the brightness of the field of view by turning the dia-illumination brightness
control knob. You can also adjust the brightness with an ND filter for Ci-S.
(2) Rotate the stage knob to move the stage and bring the target into the optical path.
(3) If the specimen is not in focus, turn the focus knob to focus on it.

13

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 Chapter 1 Microscopy Procedures

16 Adjust the field diaphragm.

Turn the field diaphragm dial to adjust the field diaphragm
so that it almost circumscribes the field of view.

Size of the field diaphragm Field

diaphragm
Normally, adjust the field diaphragm so that it
dial
almost circumscribes the field of view. Opening
the field diaphragm too much results in stray light
entering the field of view, generating flare and Field Diaphragm Field of view
reducing the image contrast. In addition, the
specimen will become decolorized over a wider
area.

Chapter 1-2 Field diaphragm's adjustment timing

Be sure to adjust the field diaphragm each time
Microscopy Procedures

you change the objective. Circumscribe around the field of view

Adjusting the field diaphragm

17 View the specimen.

Rotate the stage knob to move the target. If the target is not in focus, use the focus knob to adjust the focus.

To switch to bright-field microscopy

Phase Contrast Microscopy

● Turn the condenser turret until the [A: empty] symbol comes to the front.
● Microscopy is possible with an objective of 4x or greater, but UW microscopy with the 4x objective will
cause vignetting.
● When the condenser is set to [A: empty], its performance is equivalent to that of the Abbe condenser.

18 Turn off the power.

Turn off the power switch (press to the “O” position) for the microscope. (The power LED on the front of the main
body will turn off.)

14

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 Chapter 1 Microscopy Procedures

3 Simple Polarizing Microscopy

3.1 System Configuration and Controls

This section explains an example system configuration and the controls required for simple polarizing microscopy using
the ECLIPSE Ci-S/Ci-L.
Names of components are denoted in the following manner: [Polarizer unit for simple polarization].

Diopter adjustment ring

Optical path switching
lever

[Eyepiece] Analyzer IN/OUT knob

[Tube]

[Analyzer tube for

simple polarization]
Condenser focus knob
Chapter 1-3
[Nosepiece]
Coarse focus clamp ring

Microscopy Procedures
[Objective]
Coarse focus knob

Fine focus knob

[Stage]

Aperture diaphragm lever Y knob

Power switch
TO RQ

[Condenser]
UE

P
CL AM

[Polarizer unit for ND4 ND

8

ND filter IN/OUT switch

OUT

simple polarization]
(equipped with Ci-S only)

Simple Polarizing Microscopy

IN

[Main body] X knob

(The figure shows
ECLIPSE Ci-S.) Capture button

Field diaphragm dial

Moving claw of the

specimen holder

Condenser focus knob

Grip
Condenser centering
screw

NIKON
CORP

Input voltage
TOKYO ORATI

Coarse focus torque

, JAPAN ON MODEL
100–24 ECLIPSE
0V~ Ci-S
0.9A
50/60H
z 9400
This
device MADE 01
Rules. complie IN CHINA
Operati s with
conditio on is Part 15
ns: subject of the
(1) this to the FCC

label
4N75 device followin

adjustment knob
INSPEC and (2) may g two
this device not cause
EQUIP TION receive
must harmfu
MENT d, l interfer
undesir includin g accept
any interfer ence,
ed operati interfer
ence
This on. that may ence
Class cause
Canadi A digital
an appara
Cet appareICES-0 03. tus complie
confirm il numériq s with
e à la ue de
norme la classe
NMB-00 A est
3 du
Canada
.

Coarse focus knob

90

Lamphouse
0
80
10
70

20
60

30
50

40

AC inlet DS C Fine focus knob

Dia-illumination
DSC connector brightness control knob

15

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 Chapter 1 Microscopy Procedures

3.2 Simple Polarizing Microscopy Procedure

In this procedure, only step titles are shown for operations that are the same as those of bright-field microscopy. See “2.1
Bright-field Microscopy” for details.

1. Turn on the power. 8

2. Lower condenser slightly from
uppermost position.
17
3. Fully open field and aperture
diaphragms. 6 12
4. Bring the 10x objective into the optical 9
path.
5. Bring specimen into optical path.
6. Remove the analyzer from the optical
path. 4 14
2 10
7. Focus on specimen.
Chapter 1-3 8. Adjust diopter. 7 15
9. Adjust interpupillary. 3
Microscopy Procedures

10. Focus and center condenser. 1 18

10

TO RQ
UE
11. Bring the portion without sample into

P
CL AM
ND4 ND8
OUT

the optical path.

6 12 13
IN

12. Install polarizer unit.

13. Adjust orientation of analyzer and 5 11
polarizer. 3 16
14. Bring the desired objective into the
optical path.
Simple Polarizing Microscopy

15. Focus on specimen.

16. Circumscribe field diaphragm to field
of view.
17. View specimen.
18. Turn off the power.

Preparation for microscopy

1 Turn on the power.

(The power LED on the front of the main body will light
up.)

2 Lower the condenser slightly from the uppermost position.

3 Fully open the field diaphragm and aperture

diaphragm.

4 Bring the 10x objective into the optical path.

16

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 Chapter 1 Microscopy Procedures

5 Place a specimen on the stage, and move the

stage to bring the target into view.

6 Remove the analyzer and the polarizer from the

optical path.
Pull out the analyzer IN/OUT knob from the intermediate
tube with simple analyzer to remove the analyzer from the
optical path.
The polarizer unit for simple polarization has not yet been
installed at this point.

Remove the analyzer from the optical path.

7 Focus on the specimen. (→See Chapter 2 “2 Focusing on the Specimen (Vertical Stage Movement)” for
details)

8 Adjust the diopter. (→See Chapter 2, “4 Adjusting the Diopter” for details) Chapter 1-3

Microscopy Procedures
9 Adjust the interpupillary distance.

10 Focus and center the condenser. (→See Chapter 2, “5 Focusing and Centering the Condenser” for details)

Simple Polarizing Microscopy

17

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 Chapter 1 Microscopy Procedures

Microscopy operation

11 Bring a portion of the specimen where there is

no sample into the optical path.
Rotate the stage knob to move the specimen and bring a portion where there is no sample under the cover glass into
the optical path.

12 Bring the analyzer and the polarizer into the

optical path.
Push in the analyzer IN/OUT knob into the analyzer tube
for simple polarization to bring the analyzer into the optical
path.
Set the polarizer unit for simple polarization over the field Analyzer
lens. IN/OUT knob
Make sure the orientation mark on the polarizer (roughly)
comes to the front at this point.
Keep the fixing screw of the polarizer unit loosened.

Chapter 1-3
Microscopy Procedures

Pivot

TO RQ
UE

P
CL AM
ND4 ND8
OUT

IN

Orientation
mark Fixing screw
Simple Polarizing Microscopy

Bringing the analyzer into the optical path

Installing the polarizer

Slider-type analyzer for simple

polarization
Using the D-SA Analyzer Slider for Simple
Polarization instead of an analyzer tube for simple
polarization can keep the eye point from rising. To
use the D-SA Analyzer Slider for Simple D-SA Analyzer
Polarization, the C-NA sextuple nosepiece with slider
analyzer slot is required. Use the analyzer slider
as follows:
Push in: The analyzer goes into the optical path.
Pull out: The analyzer is removed from and the Using D-SA analyzer slider
dummy hole goes into the optical path.

18

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 Chapter 1 Microscopy Procedures

13 Adjust the orientation of the analyzer and the

polarizer.
(1) Fully open the aperture diaphragm.
(2) Pull out one eyepiece from the tube.
(3) Look into the eyepiece sleeve and rotate the whole polarizer unit until you can identify a dark cross.
(You will see black stripes that change shape as you rotate the polarizer unit.)

(4) Tighten the fixing screw of the polarizer unit to fix the
polarizer.
(5) Place the eyepiece back into the tube.

Dark cross (crossed Nicols)

You will see a dark cross when the orientation of
the analyzer orthogonally crosses that of the Dark cross
polarizer.

14 Bring an arbitrary objective into the optical path.

Turn the nosepiece to bring the desired objective into the optical path. Chapter 1-3

Microscopy Procedures
Swing-out condenser 1-100x
Mounting a 1-100x swing-out condenser on the
Ci-L main body when using the 1x objective may
result in uneven illumination around the field of
view.

15 Focus on the specimen.

(1) Look into the eyepiece, and adjust the brightness of the field of view by turning the dia-illumination brightness
control knob. You can also adjust the brightness with an ND filter for Ci-S.

Simple Polarizing Microscopy

(2) Rotate the stage knob to move the stage and bring the target into the optical path.
(3) If the specimen is not in focus, turn the focus knob to focus on it.

16 Adjust the field diaphragm.

Turn the field diaphragm dial to adjust the field diaphragm
so that it almost circumscribes the field of view.

Size of the field diaphragm Field

diaphragm
Normally, adjust the field diaphragm so that it dial
almost circumscribes the field of view. Opening
the field diaphragm too much results in stray light
entering the field of view, generating flare and Field Diaphragm Field of view
reducing the image contrast. In addition, the
sample will become decolorized over a wider
area.

Field diaphragm's adjustment timing

Be sure to adjust the field diaphragm each time
you change the objective. Circumscribe around the field of view

Adjusting the field diaphragm

19

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 Chapter 1 Microscopy Procedures

17 View the specimen.

Rotate the stage knob to move the target. If the target is not in focus, use the focus knob to adjust the focus.

Strict polarizing microscopy

If you need a retardation measurement or stricter polarizing observation, use a dedicated polarizing
microscope.

To switch to bright-field microscopy

Pull out the analyzer IN/OUT knob to remove the analyzer from the optical path. Remove the polarizer unit
from the field lens.

18 Turn off the power.

Turn off the power switch (press to the “O” position) for the microscope. (The power LED on the front of the main
body will turn off.)

Chapter 1-3
Microscopy Procedures
Simple Polarizing Microscopy

20

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 Chapter 1 Microscopy Procedures

4 Sensitive Tint Plate Microscopy

4.1 System Configuration and Controls

This section explains an example system configuration and the controls required for sensitive polarization microscopy
using the ECLIPSE Ci-S/Ci-L.
Names of components are denoted in the following manner: [Polarizer unit for first-order red compensation].

Diopter adjustment ring

Optical path
switching lever

[Eyepiece] Analyzer IN/OUT knob

[Tube]

[Analyzer tube for

first-order
red compensation] Condenser focus knob
[Nosepiece]

[Objective] Coarse focus clamp ring

[Stage] Coarse focus knob

Fine focus knob Chapter 1-4

Aperture diaphragm

Microscopy Procedures
lever Y knob
[Condenser]
TO RQ

Power switch
UE

[Polarizer unit for

CL AM

8
ND4 ND

first-order
OUT

ND filter IN/OUT switch

red compensation] IN
(equipped with Ci-S only)
[Main unit] X knob
(The figure shows
ECLIPSE Ci-S.) Capture button

Field diaphragm dial

Sensitive Tint Plate Microscopy

Moving claw of the
specimen holder

Condenser focus knob

Grip
Condenser centering
screw

NIKON

Input voltage
CORP
TOKYO ORAT

Coarse focus torque

, JAPAN ION MODEL
100–24 ECLIPSE
0V~ Ci-S
0.9A
50/60H
z 9400
This
device MADE 01
Rules. complie IN CHINA
Operati s with
conditio on is Part 15
ns: subject of the
(1) this to the FCC

label
4N75 device followin

adjustment knob
INSPEC and (2) may g two
this device not cause
EQUIP TION receive
must harmfu
MENT d, l interfer
undesir includin g accept
any interfer ence,
ed operati interfer
ence
This on. that may ence
Class cause
Canadi A digital
an appara
Cet appareICES-0 03. tus complie
confirm il numériq s with
e à la ue de
norme la classe
NMB-00 A est
3 du
Canada
.

Coarse focus knob

90

Lamphouse
0
80
10

Fine focus knob

70

20
60

30
50

40

AC inlet DS C

Dia-illumination
DSC connector brightness control knob

21

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 Chapter 1 Microscopy Procedures

4.2 Sensitive Tint Plate Microscopy Procedure

In this procedure, only step titles are shown for operations that are the same as those of bright-field microscopy. See “2.1
Bright-field Microscopy” for details.

8
1. Turn on the power.
2. Lower condenser slightly from
uppermost position. 17
3. Fully open field and aperture 6 12
diaphragms. 9
4. Bring the 10x objective into the optical
path.
5. Bring specimen into optical path.
6. Remove the analyzer from the optical 4 14
path. 2 10
7. Focus on specimen.
7 15
8. Adjust diopter. 3
9. Adjust interpupillary. 1 18
10

TO RQ
UE
10. Focus and center condenser.

P
CL AM
ND4 ND8
OUT

Chapter 1-4 11. Bring the portion without sample into

6 12 13
IN

the optical path.

Microscopy Procedures

12. Install polarizer unit.

5 11
13. Adjust orientation of analyzer and 3 16
polarizer.
14. Bring the desired objective into the
optical path.
15. Focus on specimen.
16. Circumscribe field diaphragm to field
of view.
Sensitive Tint Plate Microscopy

17. View specimen.

18. Turn off the power.

Preparation for microscopy

1 Turn on the power.

(The power LED on the front of the main body will light
up.)

2 Lower the condenser slightly from the uppermost position.

3 Fully open the field diaphragm and aperture

diaphragm.

4 Bring the 10x objective into the optical path.

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 Chapter 1 Microscopy Procedures

5 Place a specimen on the stage, and move the

stage to bring the target into view.

6 Remove the analyzer and the polarizer from the

optical path.
Pull out the analyzer IN/OUT knob from the sensitive tint
plate analyzer unit to remove the analyzer from the optical
path.
The polarizer unit for first-order red compensation has not
been installed at this point.

Removing the analyzer from the optical path

7 Focus on the specimen. (→See Chapter 2 “2 Focusing on the Specimen (Vertical Stage Movement)” for
details)

8 Adjust the diopter. (→See Chapter 2, “4 Adjusting the Diopter” for details)

9 Adjust the interpupillary distance.

Chapter 1-4

10 Focus and center the condenser. (→See Chapter 2, “5 Focusing and Centering the Condenser” for details)

Microscopy Procedures
Sensitive Tint Plate Microscopy

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 Chapter 1 Microscopy Procedures

Microscopy operation

11 Bring a portion of the specimen where there is

no sample into the optical path.
Rotate the stage knob to move the specimen and bring a portion where there is no sample under the cover glass
into the optical path.

12 Bring the analyzer and the polarizer into the

optical path.
Push in the analyzer IN/OUT knob into the analyzer tube
for first-order red compensation to bring the analyzer into
the optical path.
Set the polarizer unit for first-order red compensation over Analyzer
the field lens. IN/OUT knob
Make sure the orientation mark on the polarizer (roughly)
comes to the front at this point.
Keep the fixing screw of the polarizer unit loosened.

Pivot
Chapter 1-4

TO RQ
Orientation mark

UE
Microscopy Procedures

P
CL AM
ND4 ND8
OUT

Tint plate
rotation
IN

lever
Fixing screw

Bringing the analyzer into the optical path

Installing the polarizer

Slider-type analyzer for first-order red

Sensitive Tint Plate Microscopy

compensation
Using the C-AS Analyzer Slider for First-order Red
Compensation instead of an analyzer tube for
first-order red compensation can keep the eye
point from rising. To use the C-AS Analyzer Slider C-AS analyzer
slider
for First-order Red Compensation, the C-NA
sextuple nosepiece with analyzer slot is required.
Use the analyzer slider as follows:
Push in: The analyzer goes into the optical path.
Pull out: The analyzer is removed from and the Using D-AS analyzer slider
dummy hole goes into the optical path.

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 Chapter 1 Microscopy Procedures

13 Adjust the orientation of the analyzer and the

polarizer.
(1) Fully open the aperture diaphragm.
(2) Pull out one eyepiece from the tube.
(3) Rotate the lambda plate rotation lever (which makes the upper polarizer move around the pivot) to remove the
lambda plate from the optical path. When doing this, support the polarizer unit for first-order red compensation
with hands so that it does not move.
(4) Look into the eyepiece sleeve and rotate the whole polarizer unit until you can identify a dark cross.
(You will see black stripes that change shape as you rotate the polarizer unit.)
(5) Tighten the fixing screw of the polarizer unit to fix the polarizer.
(6) Put the eyepiece back to the tube and set the lambda plate back into the optical path.
(7) Rotate the lambda plate rotation lever to the left/right
limit to make sure that the field of view is colored
magenta on both sides.

Dark cross (crossed Nicols)

You will see a dark cross when the orientation of
the analyzer orthogonally crosses that of the Dark cross
polarizer.

14 Bring an arbitrary objective into the optical path.

Turn the nosepiece to bring the desired objective into the optical path.
Chapter 1-4
Swing-out condenser 1-100x

Microscopy Procedures
Mounting a 1-100x swing-out condenser on the
Ci-L main body when using the 1x objective may
result in uneven illumination around the field of
view.

15 Focus on the specimen.

(1) Look into the eyepiece, and adjust the brightness of the field of view by turning the dia-illumination brightness
control knob. You can also adjust the brightness with an ND filter for Ci-S.
(2) Rotate the stage knob to move the stage and bring the target into the optical path.

Sensitive Tint Plate Microscopy

(3) If the specimen is not in focus, turn the focus knob to focus on it.

16 Adjust the field diaphragm.

Turn the field diaphragm dial to adjust the field diaphragm
so that it almost circumscribes the field of view.

Size of the field diaphragm Field

Normally, adjust the field diaphragm so that it diaphragm
almost circumscribes the field of view. Opening dial
the field diaphragm too much results in stray light
entering the field of view, generating flare and
Field diaphragm Field of view
reducing the image contrast. In addition, the
sample will become decolorized over a wider
area.

Field diaphragm's adjustment timing

Be sure to adjust the field diaphragm each time
you change the objective.
Circumscribe around the field of view

Adjusting the field diaphragm

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 Chapter 1 Microscopy Procedures

17 View the specimen.

(1) Swing out the lambda plate from the optical path. (The field of view gets darker.)
(2) Rotate the stage knob to move the target. If the target is not in focus, use the focus knob to adjust the focus.
(The specimen looks brighter in the dark field of view.)
(3) Bring the lambda plate back into the optical path. (The background of the field of view will be colored
magenta.)
(4) Of the needle-like crystals seen in the field of view, check the color of the longitudinal ones.
(5) Turn the lambda plate rotation lever from right to left (clockwise) to check the change of color of the crystal
being observed.
Identify the crystal by its change of color. (See the table below.)

Position of the lambda plate rotation lever

Crystal
Leftmost Rightmost

Vibration Vibration
Crystal Crystal direction of
direction of
(yellow) the polarizer (blue) the polarizer

Urate crystal
Vibration Vibration
Z’ direction of
direction of Z’
Direction of the the analyzer Direction of the the analyzer
lambda plate lambda plate
Chapter 1-4
Microscopy Procedures

Vibration Crystal Vibration

Crystal direction of (yellow) direction of
(blue) the polarizer the polarizer
Calcium
pyrophosphoric
acid crystal
Vibration Vibration
Z’ direction of Z’ direction of
Direction of the the analyzer the analyzer
Direction of the
lambda plate lambda plate
Sensitive Tint Plate Microscopy

Keep the lambda plate clean

Note that dirt such as dust and fingerprint on the lambda plate can significantly degrade the polarization
performance. Keep it clean.

To switch to bright-field microscopy

Pull out the analyzer IN/OUT knob to remove the analyzer from the optical path. Remove the polarizer unit
from the field lens.

18 Turn off the power.

Turn off the power switch (press to the “O” position) for the microscope. (The power LED on the front of the main
body will turn off.)

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 Chapter 1 Microscopy Procedures

5 Epi-fluorescence Microscopy

5.1 System Configuration and Controls

This section explains an example system configuration and the controls required for epi-fluorescence microscopy using
the ECLIPSE Ci-S/Ci-L.
Names of components are denoted in the following manner: [CI-FL epi-fluorescence attachment].

Diopter adjustment
ring Optical path switching lever
[Eyepiece]
Filter cube switching knob
[Tube] Field diaphragm lever
[HG fiber]
Filter cube motion
restricting lever
[HG adapter]
[CI-FL epi-fluorescence B
C
A
1-2-
1-2 /
3-4
3-4
4

1-2-
3-4

attachment] CUBE
1
Condenser focus knob
Shutter open/close
1 2 3 4

lever Coarse focus clamp ring

[Nosepiece]
Coarse focus
knob
[Objective] INTENS
ILIG
C- HG HT
FI
SHUTTE
R

Fine focus
[Stage] knob
Aperture
TO RQ

ND
UE

Power switch
4
16

diaphragm lever
2
P
CL AM

32
1
ND4 ND8
OUT

[Condenser] IN

LAMP

[Main unit] ND filter IN/OUT switch RU N

TIM E

(The figure shows Chapter 1-5

hrs .

(equipped with Ci-S only)

POWER

ECLIPSE Ci-S.)
Y knob

Microscopy Procedures
Capture button
Field diaphragm dial X knob [HG precentered fiber illuminator]
(For details, see the instruction manual
for the HG precentered fiber illuminator.)

Filter cube replacement

Epi-fluorescence Microscopy
cover
ND filter slider
Light shielding plate

Moving claw of the

specimen holder
Condenser focus knob

Grip Condenser centering

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adjustment knob
Lamphouse 80
90
0
Coarse focus knob
10
70

20
60

30

Fine focus knob

50

40

AC inlet DS C

DSC connector Dia-illumination

brightness control knob

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 Chapter 1 Microscopy Procedures

5.2 Epi-fluorescence Microscopy Procedure

WARNING
The light source used with the epi-fluorescence attachment (mercury lamp) requires special care during handling
because of its characteristics. Make sure you are familiar with and adhere to all warnings and cautions described at
the beginning of this instruction manual.

Locate the observation target on the specimen under bright-field microscopy, then proceed to epi-fluorescence
microscopy.
(See Chapter 2, “15 Tips for Epi-fluorescence Microscopy” for the tips to locate the observation target on the specimen.)

1. Turn off the dia-illumination power.

2. Close the shutter.
3. Bring the filter cube into the optical 4 9
path.
4. Fully open the field diaphragm. 10
5. Turn on the mercury lamp.
A
1-2-
B 3-4

6. Open the shutter.

1-2 /
C 3-4

4
1-2-
3-4

CUBE
1

7. Bring the desired objective into the 2 6

3
1 2 3 4

optical path.
8. Focus on specimen.
9. Circumscribe field diaphragm to
7
8
INTENSIL
C- HGIGHT
FI
SHUTTE
R

field of view.
10. View specimen. 1
TO RQ

ND
UE

8
4
16

11. Turn off the mercury lamp.

2
P
CL AM

32
1

Chapter 1-5
ND4 ND8
OUT

IN
Microscopy Procedures

LAMP

RUN
TIM E
hrs .

POWER

5 11

Microscopy operation (→See also: Chapter 2, “15 Tips for Epi-fluorescence Microscopy”)
Epi-fluorescence Microscopy

1 Turn off the microscope power switch (turn off the dia-illumination).
Turn off the power switch (press to the “O” position) for the
microscope. (The power LED on the front of the main
body will turn off.) 8
ND4 ND
OUT

IN

Turning off the microscope

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 Chapter 1 Microscopy Procedures

2 Close the shutter and block the illumination

path.
Set the shutter open/close lever of the epi-fluorescence
Filter cube
attachment to position “C” to close the shutter and block switching knob
the optical path.

A
1-2-
B 3-4
1-2 /
C 3-4

4
Shutter of the epi-fluorescence attachment
1-2-
3-4

CUBE
1

The shutter blocks illumination.

1 2 3 4

If the specimen is continuously exposed to the strong

light of the mercury lamp, it may become damaged or Filter cube
Shutter closed (C)
decolorized. position nameplate window
Be sure to close the shutter when suspending the Closing the shutter
microscopy or when pausing epi-fluorescence Bringing the filter cube into the optical path
microscopy to perform microscopy with diascopic
light. Be sure to get into the habit of performing this
operation.

3 Bring the filter cube into the optical path.

Bring the desired cube into the optical path by turning the
filter cube switching knob.
(Confirm the address of the filter cube to be used, and
match the switching knob with the number.)

Selecting a filter cube

A filter cube consists of three types of optical
components: an excitation filter (EX filter), a Chapter 1-5
barrier filter (BA filter), and a dichroic mirror (DM).

Microscopy Procedures
Select the filter cube with the appropriate
combination of optical components for the
characteristics of the specimen and the
fluorescence dye.

4 Fully open the field diaphragm

epi-fluorescence attachment.
of the

Push in the field diaphragm lever of the epi-fluorescence

attachment to open the diaphragm fully.
Field diaphragm

5 Epi-fluorescence Microscopy
lever
Turn on the mercury lamp.
See your illuminator's manual for details.

6
A
1-2-
B 3-4
1-2 /
C 3-4
4

1-2-
3-4

Open the shutter. CUBE

1

1 2 3 4

Set the shutter open/close lever of the epi-fluorescence

attachment to position “O” to open the shutter.
Shutter opened Light shielding plate
Light shielding plate of the (O) position
epi-fluorescence attachment Fully opening the field diaphragm
The light shielding plate protects the observer's Opening the shutter
eyes from reflected ultraviolet light, which is
originally emitted from the objective at the
specimen.

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 Chapter 1 Microscopy Procedures

7 Bring an arbitrary objective into the optical path.

Turn the nosepiece to bring the desired objective into the optical path.
When using an oil immersion type objective, apply immersion oil between the specimen and the objective.
(→Chapter 2 “12 Oil Immersion” for details)

Non-fluorescent immersion oil

Use Nikon designated non-fluorescent immersion oil.

8 Focus on the specimen.

(1) Look into the eyepiece, and adjust the brightness of

the field of view with the ND filter of the
epi-fluorescence attachment.
(2) Rotate the stage knob to move the stage and bring
the target into the optical path.
(3) If the specimen is not in focus, turn the focus knob to ND4
ND8
ND16
A
B
1-2-
3-4

1-2 /
3-4
3-4
1-2-
C

focus on it.

Adjusting the brightness with ND filters

9 Adjust the field diaphragm of the

epi-fluorescence attachment.
Use the field diaphragm lever of the epi-fluorescence
attachment to adjust the field diaphragm so that it almost Field diaphragm
lever
circumscribes the field of view.
Chapter 1-5
Size of the field diaphragm
Microscopy Procedures

Normally, adjust the field diaphragm so that it B

C
A
1-2-
1-2 /
3-4
3-4
4

1-2-
3-4

almost circumscribes the field of view. Opening CUBE

1

the field diaphragm too much results in stray light

1 2 3 4

entering the field of view, generating flare and

reducing the image contrast. In addition, the
specimen will become decolorized over a wider
area.

Field diaphragm Field of view

Field diaphragm's adjustment timing
Be sure to adjust the field diaphragm each time
Epi-fluorescence Microscopy

you change the objective.

Circumscribe around the field of view

Adjusting the field diaphragm

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 Chapter 1 Microscopy Procedures

10 View the specimen.

Rotate the stage knob to move the target. If the target is
not in focus, use the focus knob to adjust the focus.

Diascopic image in fluorescence observation

For fluorescence observations, turn off the microscope power switch to make the diascopic image disappear.
Bright ambient lights will make it difficult to view the image. Nikon recommends keeping the room dark during
fluorescence observations.

To return to bright-field microscopy

● Close the shutter of the epi-fluorescence attachment and block the light emitted by the mercury lamp.
● Turn on the microscope power switch to turn on the dia-illumination lamp.
● Turn the filter cube switching knob and bring the position where a filter cube is not located into the optical
path.

11 Turn off the mercury lamp.

See your illuminator's manual for details.

Microscope power switch

If the microscopy power switch has been turned back ON for bright-field microscopy operation, make sure to
turn off the power switch (press to the “O” position). (Make sure that the power LED on the front of the main
body is turned off.)

Chapter 1-5

Microscopy Procedures
Epi-fluorescence Microscopy

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 Chapter 1 Microscopy Procedures

Chapter 1-5
Microscopy Procedures
Epi-fluorescence Microscopy

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 2 Individual Operations

1 Adjusting the Brightness of a Diascopic Image

The brightness of a diascopic image can be adjusted by changing the voltage of the lamp with the dia-illumination
brightness control knob or by removing/inserting ND filters from or into the optical path.

1.1 Adjustment with the Dia-illumination Brightness Control Knob

Turning the dia-illumination brightness control knob changes the

voltage of the lamp/LED to change the brightness of the
diascopic image. The brightness control knob can be used with

MP
C LA
click-less operability. 80

TO R
70 90

QU
60

E
0
Brightness Control Knob Rotation and Brightness of the

50

10
Image
0
4

20
30

Brighter
Darker
POWER

Brightness control knob Image brightness

Clockwise rotation Brighter

Dia-illumination brightness
Counterclockwise rotation Darker control knob
Adjusting the dia-illumination lamp

When using Ci-S (to retain the color balance of the image)
Adjusting brightness with the brightness control knob will affect the lamp color temperature and alter the color balance
of the image. If accurate color reproduction is critical, set the brightness control knob to the position and use the
ND filters for brightness adjustments.

1.2 Adjustment with the ND Filter (for Ci-S)

ND filters are used to adjust light intensity. Higher filter numbers

correspond to lower transmittance (i.e., darker images). The color
balance of the image will not change. Chapter 2
Ci-S has built-in ND filters (ND4 and ND8).

Individual Operations
Pushing the each filter IN/OUT switch inserts the corresponding
TO RQ
UE

ND filter into the optical path.

CL AM

ND4 ND8
OUT

ND4: Reduces light intensity to 1/4. IN

ND8: Reduces light intensity to 1/8.

Filter cassette ND filter IN/OUT
ND4+ND8: Reduces light intensity to 1/32. holder switch

Adjusting the brightness with ND filters

Placing the ND filers over the field lens

You can add an ND filter by placing the ND filters over the field lens or mounting the filter cassette holder onto the field
lens part.
The filter cassette holder can house up to three φ45 mm filters with a thickness of up to 3 mm. Pushing in the filter
IN/OUT lever inserts the ND filters into the optical path. In addition, one filter can be placed on the filter cassette holder.
The filter cassette holder cannot be used with a simple polarizer, polarizer unit for first-order red compensation, or the
spacer for the nosepiece.

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 Chapter 2 Individual Operations

1.3 Removing an NCB filter for a brighter image (Ci-S)

Ci-S has an integrated NCB filter (NCB 11) to improve the color reproduction.
If the image still looks dark even after all ND filters are removed from the optical path, removing this NCB filter can make the
image brighter.
Follow the procedure below to remove the NCB filter: NCB filter slot cover fixing screws
(1) Turn off the power switches for the microscope main body
and the peripheral devices (press to the “O” position) and
remove the power cord from the receptacle.
(2) Wait until the lamp and the peripheral devices cool down
sufficiently (about thirty minutes).
(3) Remove all accessories such as the camera, objective,
nosepiece, eyepiece, tube, condenser, and the stage. See
Chapter 3 “Assembly” and use a reverse order of assembling
to remove the accessories.
(4) Place the microscope main body upside down with its bottom
facing up.
(5) Loosen two NCB filter slot cover fixing screws on the bottom
with a tool, remove the cover, and remove the NCB filter
along with the filter holder.
(6) Restore the microscope main body in place and follow the
procedure in Chapter 3 “Assembly” to attach those Front
accessories back in place.
Remove the NCB filter

Chapter 2
Individual Operations

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 Chapter 2 Individual Operations

2 Focusing on the Specimen (Vertical Stage Movement)

Note on controlling the focus knobs

Avoid the following actions, which can cause equipment
malfunction.
0.8 0.6 0.4 0.2

• Rotating the right and left focus knobs in opposite directions.

• Rotating the coarse focus knob past the limit.

POWER

Don't rotate the knobs in opposite directions!

Turn the coarse or fine focus knob to raise or lower the stage and
10x
shift the focus onto the specimen.
Using an objective of high magnification may cause the specimen
to be pushed against the objective, damaging the objective.
Follow the procedure below to focus on the specimen to avoid

P
CL AM
breaking the cover glass or damaging the objective. 80

TO RQ
70 90

UE
60

0
50
(1) Bring the 10x objective into the optical path and turn the

10
40

20
30

coarse focus knob away to raise the stage to the upper limit.
POWER

Darker Brighter
(1)
(2) When using the trinocular tube or ergonomic tube, push in Select the 10x objective
the optical path switching lever to distribute 100% light to the and use the coarse (3)
binocular section. focus knob to raise the Adjust the brightness
stage to the upper limit. of the field of view.
(3) Look into the eyepiece, and adjust the brightness of the field
Bringing the 10x objective into the optical path,
of view by turning the dia-illumination brightness control and raising the stage to the upper limit
knob. You can adjust the brightness with an ND filter for Ci-S.
(4) Look into the eyepiece and turn the coarse focus knob slowly
to lower the stage and focus on the specimen. Release your
hand from the coarse focus knob once it has been focused.
(5) Turn the fine focus knob to adjust the focus more accurately. (4) Chapter 2
Use the coarse
P
CL AM

focus knob to focus.

80

Individual Operations
TO RQ

70 90
UE
60

0
50

(5)
10

40
20

30

POWER

Use the fine focus

knob for more
accurate focus.
Using the coarse focus knob to focus→Using the
fine focus knob for more accurate focus

Raising the stage with the coarse focus knob

When moving the stage with the coarse focus knob, move your eyes away from the eyepiece and operate the
microscope while looking at it from the side.

Looking into the eyepiece during coarse operation

When working with the coarse focus knob while looking into the eyepiece, you should only turn the knob in the direction
for lowering the stage.

Switching to an objective of higher magnification

First use an objective of low magnification to adjust the focus, then switch to an objective of higher magnification.

Operating distance
As a 10x or 4x objective has a wider operating distance, the specimen never touches the tip of the objective even if the
stage is raised to the upper limit, provided that a slide and a cover glass of standard thickness are used. (Standard
thickness being 1.2 mm for the slide glass, and 0.17 mm for the cover glass.)

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 Chapter 2 Individual Operations

2.1 Focus Knob Rotation and Stage Movement

Both the coarse focus knob and the fine focus knob are located
on both right and left sides on the microscope. The table below
shows the relationship of the focus knobs' rotation with the stage
movement.

Focus Knob Rotation and Stage Movement

P
CL AM
80

TO RQ
70 90

Stage is lowered.

UE
60
Operations Stage movement

0
50

10
40

20
30

POWER

Turn the knob toward the Stage is lowered.

front.
Fine focus Coarse focus
Turn the knob toward the Stage is raised. knob knob
rear.
Stage Vertical Movement

2.2 Number of Focus Knob Turns and Distance of Stage Travel

Number of Focus Knob Turns and Distance of Stage Travel

Distance of stage travel

No. of knob turns
(vertical direction)

One rotation of the coarse focus knob Approx. 9.33 mm

One rotation of the fine focus knob Approx. 0.1 mm

One scale of the fine focus knob 1 µm

The vertical motion range (coarse/fine focus stroke) of the stage is from approximately 2 mm above the focal point
(reference position) to approximately 28 mm below the focal point.

Chapter 2
Individual Operations

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 Chapter 2 Individual Operations

2.3 Adjusting the Rotating Torque of the Coarse Focus Knob

Adjust the rotation torque of the coarse focus knob (rotation

resistance) by turning the torque adjustment knob (TORQUE)
located at the base of the coarse focus knob. If the torque is set
too low, the stage may descend under its own weight.
Makes knob
Adjusting the Rotating Torque of the Coarse Focus Knob harder to turn.

P
CL AM
80

TO RQ
70 90

UE
60
Operation of torque Rotation torque

0
50

10
40

20
30

adjustment knob POWER

When turned in the Rotation torque is increased.

direction of the arrow Coarse focus torque adjustment knob

When turned in the Rotation torque is decreased.

direction opposite to the Adjusting the torque of the focus knobs
arrow

2.4 Refocusing

By turning the coarse focus clamp ring after focusing on the

specimen, you can prevent the stage from being raised further
with the coarse focus knob. The movement of the stage with the
fine focus knob will not be locked.

TO RQ
UE

P
CL AM
Using this function, you can refocus with ease by simply turning OUT
ND4 ND8

the coarse focus knob to the limit. This is helpful when switching IN

between similar specimens during the observation.

(1) With the focus set on the specimen, tighten the coarse focus Coarse focus clamp ring
clamp ring by turning it approximately 3/4 of a rotation in the
direction of the arrow on the base of the microscope. This will Refocusing
clamp the movement of the coarse focus knob.
(2) When replacing the specimen, lower the stage by using only
the coarse focus knob.
(3) After replacing the specimen, use only the coarse focus knob Chapter 2
to raise the stage slowly until it reaches the upper limit.

Individual Operations
At the upper limit, the focus should be more or less on the
specimen. Use the fine focus knob for finer adjustment.
If you do not wish to use the refocusing function, be sure to
loosen the coarse focus clamp ring to the limit (turn it in the
direction opposite to the arrow on the base of the
microscope until it hits the limit).

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 Chapter 2 Individual Operations

2.5 Position Exchange of the Fine Focus Knob

Among left and right fine focus knobs, one is flat and another is
convex. Both fine focus knobs are attached to the coarse focus 0.8 0.6 0.4 0.2

knobs using magnet, so you can detach the left and right knobs
from the coarse focus knobs and swap them.
Position them to best suit your usage.

Removing a flat focus knob

A flat fine focus knob can be easily removed by inserting a POWER

flathead screwdriver or the likes into the notch in the knob.

Convex fine focus knob Flat fine focus knob

Position Exchange of the Fine Focus Knob

Chapter 2
Individual Operations

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 Chapter 2 Individual Operations

3 Bringing the specimen into the optical path (Horizontal Stage Movement)

Note on moving the stage

Avoid the following action, which can cause equipment
malfunction.
• Moving the stage to the right and left by holding the top
surface of the stage directly. 0.8 0.6 0.4 0.2

Do not move the top surface of the stage by hand!

Turning the stage knob moves the upper plate of the stage in the X and Y directions so that you can move the stage to bring
the target into the optical path.
This illuminates the sample sealed under the cover glass.

3.1 Knob Rotation and Stage Movement

Y direction X direction
To move the stage in the X or Y direction, rotate the X knob or Y
knob.

TO RQ
UE

P
CL AM
ND4 ND8
OUT

IN

Y knob X knob

XY Stage Movement Chapter 2

Individual Operations
3.2 Adjusting the Knob Heights

The heights (positions) of the X knob and Y knob can be changed. Hold the knob and move it along the vertical axis to the
desired height.

3.3 Adjusting the Knob Rotation Torque

When the X knob and Y knob are moved to the top and bottom Rotation in the direction
indicated by the arrow
positions, the torque adjustment screws can be found between increases the torque.
the knobs. Turning the torque adjustment screw to move it closer
0.8 0.6 0.4 0.2

to the respective knob increases rotational torque. (To increase

the rotational torque, turn the adjustment screw counterclockwise
for the Y knob and clockwise for the X knob, as viewed from
above.)
Avoid loosening these screws excessively. If they are too loose,
the top surface of the stage may move, even when touched very
Y knob torque X knob torque
lightly. adjustment screw
adjustment screw
Adjusting the torque of XY knobs

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 Chapter 2 Individual Operations

4 Adjusting the Diopter

The diopter adjustment ring on an eyepiece can be adjusted to match the eyesight of your right and left eyes.
A properly adjusted diopter compensates for differences in visual acuity between the right and left eyes of a person,
making binocular observation easier. It also minimizes focal deviations when switching magnification, optimizing the
performance of the objective.
Adjust the diopter settings for both eyepieces.

Notch on eyepiece
The eyepiece has a notch to prevent the rotation. When attaching, match the notch with the protrusion on the eyepiece
sleeve. Otherwise, eyepiece is not attached to the correct position.

(1) Turn the diopter adjustment ring on the right and left
eyepieces to align the end face of the diopter adjustment ring (1)
with the line. (This is the diopter adjustment reference Line
position.)
(2) Follow Steps 1 through 6 in Chapter 1 “1.2 Bright-field
Microscopy” to focus on the specimen using the 10x
objective.
(3) Turn the nosepiece to bring the 40x objective into the optical
Reference position for diopter adjustment
path, and turn the coarse focus knob and then the fine focus
knob to focus on the specimen.
(4) Bring the 10x (or 4x) objective into the optical path. (3)
Select 40x.
(5) Look into the left eyepiece with your left eye. Without
touching the focus knob, focus on the specimen by turning
the left diopter adjustment ring.
(6) Look into the right eyepiece with your right eye without

P
CL AM
touching the focus knob to focus on the specimen by turning
80

TO RQ
70 90

UE
60

the right diopter adjustment ring.

50

10

40
20

30

POWER

(7) Repeat Steps (3) through (6) to make sure the focus has
Use the focus knob
been adjusted properly. to focus.

Chapter 2 (4)
Select 10x or 4x.
Individual Operations

(5) (6)
Look into the left eyepiece Also look into the right
with your left eye to focus eyepiece with your right eye
with the left diopter to focus with the right
adjustment ring. diopter adjustment ring.

Adjusting the Diopter

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 Chapter 2 Individual Operations

5 Adjusting aperture diaphragm

Adjust the condenser position so that the light passing through the condenser forms an image at the correct position
(center of the optical path) on the surface of the specimen.

(1) Follow Steps 1 through 6 in Chapter 1 “1.2 Bright-field

Microscopy” to focus on the specimen using the 10x (3)
Focus the
objective. field
(2) Turn the field diaphragm dial counterclockwise fully to diaphragm
image at 10x.
minimize the field diaphragm.

TO RQ
UE
(3) You can see the diaphragm image in the field of view as you

P
CL AM
ND4 ND8

look into the eyepiece. Use the condenser focus knob to

OUT

adjust so that the field diaphragm image can be outlined

IN

clearly. (2)
Minimize the field
(4) Adjust the condenser centering screws until the field diaphragm.
diaphragm image is at the center of the eyepiece field of
view.
(5) Set the 40x objective into the optical path. Bring the 40x 0.8 0.6 0.4 0.2

objective into the optical path. As you see the field diaphragm
image, if the outline of the image is out of focus, use the
condenser focus knob to adjust the focus as much as
possible.
(6) Turn the field diaphragm dial to adjust so that the field
POWER

diaphragm image size is almost the same as the field of view. (4)
Field diaphragm image
(7) When the center of the field diaphragm image is not centered at the field of view
centered, turn the condenser centering screws to move the
field diaphragm image to the center of the field of view. This (5)
is easiest if you adjust the field diaphragm aperture so that it Focus the
is slightly smaller than the eyepiece field of view. field
diaphragm
Correct focusing of the field diaphragm image accurately at
40x.
TO RQ

If the outline of the field diaphragm image is reddish or

UE

P
CL AM

bluish, you have turned the condenser focus knob too OUT
ND4 ND8

much. When the outline is colorless, focusing is correct. (7)

Field Chapter 2
IN

diaphragm
Field diaphragm image view with the 40x image

Individual Operations
objective centered at
the field of (6)
The field diaphragm image that has been focused with the view
40x objective can not be seen so clearly as the one Open the field
diaphragm up to the
focused with the 10x objective. size of the field of view.

Field diaphragm Field diaphragm

image image

Eyepiece field ring Eyepiece field ring

Focusing and centering condenser

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 Chapter 2 Individual Operations

6 Adjusting the Aperture Diaphragm

The aperture diaphragm is used to adjust the illumination

angular aperture and is important as it affects the resolution,
contrast, focal depth, and brightness of an optical image.
Turning the condenser aperture diaphragm lever changes the
size of the aperture diaphragm.
Generally, aperture settings at 70 to 80% of the numerical

TO RQ
UE
aperture of the objective will provide satisfactory images with

P
CL AM
ND4 ND8
OUT

suitable contrast. IN

A small aperture diaphragm opening reduces resolution and

brightness but increases contrast and depth of focus. On the
contrary, a large aperture diaphragm size increases resolution Aperture diaphragm
and brightness but reduces contrast and depth of focus. These lever
characteristics involve inherent tradeoffs and cannot be Adjusting the aperture diaphragm
optimized independently.

Relationship of the aperture diaphragm size with the optical image's state

Aperture diaphragm Resolution Brightness Contrast Focal depth

Stop down Lower Darker Larger Deeper
Open Higher Brighter Lesser Shallower

Proper size of the aperture diaphragm

Normally, 70 to 80% of the numerical aperture of the objective is the proper size. Since an excessively small aperture
diaphragm opening will degrade image resolution, we do not recommend setting the aperture diaphragm to less than
60% of the numerical aperture of the objective.

Adjustment timing for the aperture diaphragm

Be sure to adjust the aperture diaphragm each time you change the objective.

6.1 Adjusting the Aperture Diaphragm Using the Condenser Scale

The scale on the condenser indicates the numerical aperture. Plan 40X
Chapter 2 The index on the aperture diaphragm lever should be aligned with 40x / 0.75 Nikon JAPAN
Plan 40x

the scale line that corresponds to 70 to 80% of the numerical / -WD

40x / 0.75
/ -WD
Individual Operations

aperture of the objective.

The numerical aperture is indicated on the side of the objective. Indication for 40x magnification / numerical
For a numerical aperture of 0.75, the index should be aligned with aperture 0.75
the scale on the condenser at 0.525 to 0.6. Proper numerical aperture: 0.75 x 0.7 to 0.8=0.525 to
0.6

6.2 Adjusting the Aperture Diaphragm Using the Centering Telescope

(1) Remove one eyepiece and attach the centering telescope in

place, using the adapter. Eyepiece Flange
(2) Turn the aperture diaphragm lever to stop down to the
minimum aperture. While holding down the flange of the
centering telescope, turn the eyepiece of the centering
telescope and focus on the aperture diaphragm.
(3) Turn the aperture diaphragm lever to adjust the aperture.
Normally, the aperture diaphragm should be adjusted to
around 70 to 80% of the size of the field of view. Adjustment using the centering telescope
(4) Remove the centering telescope and adapter and reattach
the eyepiece.

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 Chapter 2 Individual Operations

7 Selecting a Condenser

Select a condenser optimal for the magnification of the objective and the microscopy procedure.

Selecting the Magnification of the Objective and Condenser

Condenser (~: Optimum, ○: Suitable, ×: Not suitable)

Slide achro
Achromat Swing-out
Objective's Swing-out Achromat Abbe condenser
aplanatic condenser
magnificatio condenser condenser condenser 2-100x
condenser 1-100x
n *4

1x × × × × ~*2 ×

2x × × ×
○*2 ~*2
4x × ○*1 ○*1 ~*3

10x to 100x ~ ○ ○ ○ ~

*1: The entire field of view may not be covered if a UW eyepiece is attached.

*2: Swing out the top lens before use. Uneven illumination around the field of view may occur.

*3: For objectives of 10x or higher magnification, pull out the slide. For objectives with 2x or 4x magnification, push in the
slide to prevent vignetting.

*4: Do not use the 2-100x slide achromat condenser with an achromat objective or a plan achromat objective.

Depending on the type of objective, the indicated numerical aperture of the objective may not be achieved.
For example, when an objective with a numerical aperture. of 1.4 is used, the maximum aperture of the swing-out
condenser or the Abbe condenser will only be about 65% of the N.A. of the objective, even when the condenser's aperture
diaphragm is wide open.
Refer to Section 14 “Tips for the Phase Contrast Microscopy” for the phase contrast condenser.

8 Adjusting the field diaphragm

Chapter 2
The field diaphragm is used to restrict illumination to the area of

Individual Operations
the specimen being viewed.
Turning the field diaphragm dial changes the size of the field
diaphragm.
TO RQ

For normal observations, the size of the diaphragm should almost

UE

P
CL AM

circumscribe the field of view. OUT

ND4 ND8

IN

Field diaphragm dial

Adjusting the field diaphragm

Proper size of the field diaphragm

Usually, the size is optimal when it almost circumscribes the field of view. Opening the field diaphragm too much and
illuminating a broader area than necessary will result in stray light entering the field of view, generating flare and
reducing the image contrast. In addition, the specimen will become decolorized over a wider area.

Field diaphragm's adjustment timing

Be sure to adjust the aperture diaphragm each time you change the objective.

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 Chapter 2 Individual Operations

9 Switching the Optical Path of the Tube

9.1 Light Distribution

With the ergonomic binocular tube or trinocular eyepiece tube,

the optical path switching lever allows distribution of light to the
binocular section and camera port.
Optical path
switching lever

Switching the Optical Path of the

Tube

Optical Path Switching Lever and Distribution of Light

Position of the optical path Light distribution (%)

switching lever Binocular section Camera port

C-TE2 Pushed in 100 0

Ergonomic tube
Pulled out 50 50

T C-TT Pushed in 100 0

trinocular tube
Pulled out by one notch 20 80

Pulled out by two notches 0 100

F C-TF Pushed in 100 0

trinocular tube
Pulled out 0 100

Chapter 2 9.2 Disabling the Clicking of the Optical Path Switching

Individual Operations

C-TT trinocular tube and F C-TF trinocular tube have a “NO

CLICK” switch on their tube attaching surface. Slide this switch in
the direction of the arrow with the tip of a pointed tool to disable
clicking for the optical path switching lever. Set the switch to this
position if you need to eliminate the slight vibrations resulting
from the clicking action. NO CLICK

Disabling the clicking of the optical path switching

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 Chapter 2 Individual Operations

10 Adjusting the Binocular Section

10.1 Adjusting with the Binocular Section of the Tube

The ergonomic tube makes it possible to tilt and extend the

binocular section. Adjust the position of the binocular section for
most comfortable viewing.

Adjusting the binocular section

10.2 Using the Eyelevel Riser

Inserting the eyelevel riser between the tube and the microscope
arm allows you to raise the eye point 25 mm higher.

Eyelevel riser
Chapter 2

Individual Operations
Using the eyelevel riser

11 Using Stage at Lowered Position (Spacer for Nosepiece)

Inserting a spacer for the nosepiece between the arm and the
nosepiece allows for operation with the stage set 20 mm lower.
A lowered stage facilitates specimen replacement during a
cytodiagnostic examination, etc.

Using the filter cassette holder not allowed

The filter cassette holder placed on the field lens is not Spacer for the
allowed to use together with the spacer for nosepiece. nosepiece

Using the spacer for the nosepiece

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 Chapter 2 Individual Operations

12 Oil Immersion

CAUTION
• Petroleum benzine and absolute alcohol are highly flammable. Be careful when handling these materials, especially
around open flames and when turning the power switch on or off.
• Follow the instructions provided by the manufacturer when using absolute alcohol.

An objective with an “Oil” marking is an oil-immersion objective.

An objective of this type is to be used with designated
non-fluorescent immersion oil applied between the sample and
the tip of the objective.
For maximum performance, an oil-immersion objective with a
numerical aperture of 1.0 or larger should be combined with an
oil-immersion achromat aplanatic condenser. The oil-immersion
condenser is used by applying a designated non-fluorescent

P
immersion oil between the sample and the condenser.

CL AM
80

TO RQ
70 90

UE
60

0
Oil immersion

Phase turret condenser

The phase turret condenser is intended to be used dry. Do not apply immersion oil between the condenser and the
specimen.

Use as little oil as possible (just enough to fill the space between the tip of the objective and the sample, or between the tip of
the condenser and the sample). Too much oil will result in excess oil flowing onto the stage and around the condenser.

Air bubbles in the oil

Any bubbles in the immersion oil will degrade image quality. Be careful to prevent bubbles from forming. To check for air
bubbles, fully open the field diaphragm and aperture diaphragm, remove the eyepiece, and examine the exit pupil
(bright round section) of the objective inside the eyepiece tube. If it is difficult to ascertain the presence of bubbles,
Chapter 2 attach a centering telescope with the adapter, then look for air bubbles while turning the eyepiece section of the
centering telescope to adjust the focus. If you detect bubbles, remove them by one of the following methods:
Individual Operations

● Turn the revolving nosepiece slightly to move the oil-immersed objective back and forth once or twice.
In the case of the condenser, gently turn the condenser focus knob to move the condenser up and down slightly.
● Add more oil.
● Remove the oil and apply new oil.

Cleaning the oil

Oil remaining on the oil-immersion objective or adhered to the dry-type objective will significantly degrade image
quality. After use, thoroughly wipe off all oil, and make sure that no oil remains on the tips of other objectives.
Additionally, carefully wipe off the oil from the condenser.
Use petroleum benzine to wipe off the immersion oil. For optimum results, Nikon recommends cleaning with absolute
alcohol (ethyl or methyl alcohol) after cleaning with petroleum benzine.
If petroleum benzine is unavailable, use methyl alcohol alone. When using just methyl alcohol, note that surfaces will
need to be wiped repeatedly to ensure complete removal of the immersion oil. (Usually, three or four times should be
sufficient to clean the lens.)

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 Chapter 2 Individual Operations

13 Water Immersion
An objective with a “WI” or “W” marking is a water-immersion objective. Such an objective is used with immersion water
(distilled water or physiological saline) applied between the specimen and the tip of the objective. Microscopy procedures
are the same as for oil-immersion objectives.
Since water evaporates readily, monitor the immersion water during observation. Avoid using too much water, since
excess water will flow onto the stage and around the condenser, promoting corrosion.

Cleaning of water
After use, wipe off the water from the tip of the objective and condenser, then follow up by wiping with absolute alcohol.
If you observe water stains, apply a small amount of neutral detergent and wipe gently, then follow up with absolute
alcohol.

14 Tips for Phase Contrast Microscopy

Phase contrast microscopy is suitable for observation of clear and colorless specimens, undyed or lightly colored
specimens, decolored specimens, and ultrathin slices for electron microscopes. The phase contrast method is not suitable
for graded or hard dyed specimens.
A phase contrast image appears differently depending on the phase contrast or shape of the specimen, and the properties
of the objective. Note the following when you prepare the specimen or select a Ph objective.

Select a specimen for which the center of the Ph annular diaphragm is not misaligned.
The center of the Ph annular diaphragm will be misaligned when observing a specimen that causes scattered light or
generates a lens or prism effect. In particular, with live and thick specimens, oversized specimens, and specimens using a
microplate, the center will be misaligned due to the lens or prism effect, making the specimen difficult to observe.

Ph objective and specimen

Ph objectives include “achromatic”, “plan achromatic”, “plan flour”, and “plan apochromatic” objectives depending on how
much the chroma aberration and field curvature are adjusted. These lenses also are further subdivided into several types
depending on the properties of the internal phase plate. For favorable microscopy results, the phase contrast amount of the
specimen must match the properties of the phase plate. See the table below for the use properties of the Ph objectives.
When using a dark contrast Ph objective, make sure that the phase contrast of the specimen does not exceed the allowed
phase contrast amount (latitude). If the phase contrast amount of the specimen is greater than the allowed phase contrast
amount, observation is not possible as the image will be illuminated brighter than the background. Chapter 2
You can increase or decrease the phase contrast by the thickness of the specimen, and the refraction index of the

Individual Operations
mounting agent or culture solution when preparing a phase contrast specimen.
A specimen with weak contrast under a DLL objective may yield better result under a DM objective.

Use Properties of Ph Objectives

Ph contrast
Appearance Contrast Latitude Usage example
objective
Phase contrast
and absorbing Bacteria's spore, general
Intermediate
DLL object live bacteria, slightly thick
Generally, an object with larger (with
Suitable for broader (chromosome) in specimen, bacteria, dyed
DL phase contrast appears darker. low and specimen, egg, fat particle,
detailed usage)
Dark Therefore, the image is shown in intermediate crystalline, etc.
observation
black in a relatively brighter field latitude
contrast mainly using
of view, similar to the one
micro High Bacteria and protozoal
observed with bright-field
contrast. (with flagellum, fibrin basic fiber,
microscopy. Transparent object
DM relatively fine granule,
in low latitude
narrower mounting-agent-selective
usage) slice, ultrathin slice, etc.
Generally, an object with larger
phase contrast appears brighter. Suitable for morphology,
Bacteria and protozoal
Bright Therefore, the image is shown detection, and calculation
flagellum, fibrin basic fiber,
BM brighter in a relatively darker of fine fiber and granule Almost all areas
contrast fine granule, blood cell
field of view, similar to the one mainly using macro
calculation, etc.
observed with dark-field contrast.
microscopy.

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 Chapter 2 Individual Operations

Using the GIF filter

The GIF filter (green interference filter) improves the contrast when placed in the optical path. The filter should be installed on
the field lens, or placed inside or on top of the filter cassette holder. Note, however, that it may cause ghosting when mounted
inside the filter cassette holder.

Phase turret condenser

Phase contrast microscopy requires a phase turret condenser equipped with a Ph annular diaphragm. Matching the Ph
annular diaphragm and the phase plate of the Ph objective correctly provides a phase contrast effect.

■ Ph code
One of the Ph codes, [Ph1], [Ph2], or [Ph3] is indicated on the Ph objective depending on the size of the phase plate.
(Ph codes have nothing to do with the magnification of the objective.) Always use a Ph objective and Ph annular
diaphragm with the same Ph code. You cannot experience the phase effect if a different combination of the codes
might be used.

■ Centering of the annular diaphragm and the phase plate

The position of each condenser diaphragm in the condenser turret have already been adjusted based on the Ph1
annular diaphragm. Usually, once centered with Ph1, further centering is not required upon switching to another
magnification. However, the phase contrast image will differ slightly depending on how the annular diaphragm
overlaps the phase plate, thus for a stricter observation or capturing of still images, check whether the annular
diaphragm and the phase plate are concentric at each magnification. In addition, slightly decentering the annular
diaphragm and the phase plate will produce a shadowing effect, resulting in a stereo image. Use this method as
appropriate for the specimen.

■ Infrared ray blocking filter (Ci-S only)

The lamphouse is equipped with an infrared ray cut filter, which reduces the infrared ray component of the
illumination to protect live specimens from being damaged by the heat from the illuminator.

Notes on using the phase turret condenser

● Microscopy is possible with an objective of 4x or greater, but UW microscopy with the 4x objective will cause
vignetting.
Chapter 2
● When the phase turret condenser is set to [A: empty], its performance is equivalent to that of the Abbe
condenser.
Individual Operations

Combining with an epi-fluorescence attachment

When the epi-fluorescence and phase contrast attachments are attached to the microscope, it is possible to use
epi-fluorescence and phase contrast microscopy in combination. This allows for compensation of each method's
shortcomings, for example, by making it possible to find the target using the phase contrast method instead of the
epi-fluorescence method that is likely to degrade the color of the specimen or by using both microscopy concurrently.
See Chapter 1 “2.2 Phase Contrast Microscopy Procedure” for the phase contrast microscopy procedure and Chapter 1
“5.2 Epi-fluorescence Microscopy Procedure” for the epi-fluorescence microscopy procedure. When switching between
microscopy methods, note the following.

To switch to epi-fluorescence microscopy

● Turn the condenser turret until the [C: Shutter] symbol comes to the front.
● Insert the desired excitation filter cube into the optical path.
● Open the shutter for the epi-fluorescence attachment.

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 Chapter 2 Individual Operations

To switch to phase contrast microscopy

● Close the shutter for the epi-fluorescence attachment and block the excitation light from the epi-fluorescence
attachment.
● Turn the filter cube switching knob and move the position without a filter cube into the optical path.
● Bring a Ph objective into the optical path.
● Turn the condenser turret to bring an annular diaphragm with the same Ph code as the objective into the optical
path.
● Fully open the aperture diaphragm.

To use epi-fluorescence and phase contrast microscopy concurrently

(1) Use phase contrast microscopy to adjust the focus onto the target.
(2) Remove the GIF filter, if installed, on the field lens.
(3) Insert the desired excitation filter cube into the optical path.
(4) Open the shutter for the epi-fluorescence attachment to focus it again.
(5) Adjust the brightness of the fluorescent image using the ND filter of the epi-fluorescence attachment.
(6) Adjust the brightness of the phase contrast image using the ND filter of the microscope. For Ci-L, use the
dia-illumination brightness control knob for adjustment.

Chapter 2

Individual Operations

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 Chapter 2 Individual Operations

15 Tips for Epi-fluorescence Microscopy

WARNING
The light source used with the epi-fluorescence attachment (mercury lamp) requires special care during handling
because of its characteristics. Make sure you are familiar with and adhere to all warnings and cautions described at
the beginning of this instruction manual.

Protecting the specimen and preventing it from decoloration

(shutter for the epi-fluorescence attachment)

The shutter blocks illumination.

Set the shutter open/close lever on the epi-fluorescence Shutter
open/close
attachment to position “C” to close the shutter and block the lever
B
C
A
1-2-
1-2 /
3-4
3-4

4
1-2-
3-4

optical path.
C: Closed
CUBE
1

If the sample is continuously exposed to the strong light of the 1 2 3 4

O: Open
mercury lamp, it may become damaged or decolorized.
Light
Be sure to close the shutter when suspending the microscopy or shielding
plate
when pausing epi-fluorescence microscopy to perform
microscopy with diascopic light. Be sure to get into the habit of Opening/closing the shutter
performing this operation.

Protecting from ultraviolet light (light shielding plate)

The light shielding plate is used to prevent the reflected ultraviolet light from entering the observer's eyes, which is originally
emitted through the objective, from the specimen.

Using a non-fluorescent slide, cover glass, and immersion oil

For fluorescence observations, be sure to use a non-fluorescent slide, cover glass and our designated immersion oil for an
image with better contrast.

Restricting the illumination to the area of the specimen being viewed

Chapter 2
(adjusting the field diaphragm)
Individual Operations

The field diaphragm is used to restrict illumination to the area of the specimen being viewed.
Turning the field diaphragm lever of the epi-fluorescence attachment changes the size of the field diaphragm.
For normal observations, stop down the diaphragm so that the aperture boundaries circumscribe or inscribe the field of
view. Opening the field diaphragm too much and illuminating a broader area than necessary will result in stray light
entering the field of view, generating flare and reducing the image contrast. In addition, the specimen will become
decolorized over a wider area.
Be sure to adjust the field diaphragm each time you change the objective.

Adjusting the brightness of the fluorescent image (adjusting the ND)

■ ND filters in the epi-fluorescence attachment

ND filters are used to adjust light intensity. Higher filter numbers correspond to lower transmittance (i.e., darker
images). ND filters do not affect the color balance.
The epi-fluorescence attachment has three ND filters (ND4, ND8, and ND16) built in.
Push in the ND filter IN/OUT levers to insert ND filters into the optical path and darken the fluorescent image.
As shown below, you can combine these three filters to achieve various levels of brightness.

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Light Reduction by Combined ND Filters of the Epi-fluorescence Attachment

Brightness ND4 ND8 ND16

1 - - -

1/4 ○ - -

1/8 - ○ -

1/16 - - ○

1/32 ○ ○ -

1/64 ○ - ○

1/128 - ○ ○

1/512 ○ ○ ○

(-: Removed from the optical path, ○: Placed into the optical path)

Improving the S/N ratio (shielding tube)

To improve the signal to noise ratio (SNR) during epi-fluorescence observations solely using epi-fluorescence
microscopy, we recommend removing the condenser and using the shielding tube provided with the epi-fluorescence
attachment.
In particular, when combining Ci-L and the epi-fluorescence attachment, excitation light from the Epi-fl attachment may
strike the white LED, making it illuminate and resulting in SNR deterioration. To avoid this situation, make use of the
shielding tube provided with the Epi-fl attachment or place a plate on the field lens.

■ ND on HG precentered fiber illuminator

You can also adjust the light intensity using the ND on the HG precentered fiber illuminator.
For detailed information, refer to the operating manual provided with the HG precentered fiber illuminator.

Locating a target on the specimen

The standard procedure for epi-fluorescence microscopy is to first locate the target under differential interference contrast
or phase contrast microscopy, and then switch to epi-fluorescence microscopy.
To locate the target under dia-illumination bright-field microscopy, you will need to note the following.
• Under dia-illumination bright-field microscopy, start with a 10x objective, and adequately stop down the condenser.
Chapter 2
• Gradually increase the magnification. When the target becomes difficult to locate, switch to epi-fluorescence, and
use low excitation light.

Individual Operations
• You can also use other techniques, such as using the edge of the cover glass to approximate the position of the
target.

15.1 Switching Excitation Methods

Four filter cubes can be inserted into the epi-fluorescence

Filter cube
attachment. (→See Chapter 3, “6 Assembly for Epi-fluorescence motion restricting
Microscopy”) B
C
A
1-2-
1-2 /
3-4
3-4
lever
4

1-2-
3-4

Move the desired cube into the optical path by turning the filter CUBE
1

Filter cube
cube switching knob on the right side of the attachment. 1 2 3 4
switching knob
For bright-field observations, leave one cube position empty, and
move this empty position into the optical path. Filter cube
nameplate window
Use the filter cube motion restricting lever located at the upper
front section to limit the cube switching operation. Switching the filter cube

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Filter Cube Motion Restricting Lever Functions

Lever position Filter cube switching

Position A
Locked
(push in by two A

(filter cubes cannot be switched)

1-2-
B 3-4
1-2 /
C 3-4

4
1-2-

notches)
3-4

CUBE
1

Switching between positions 1 and 2, or 1 2 3 4

Position B between positions 3 and 4 only.

(push in by one (The position at which the lever is pushed in
notch) determines whether switching is for positions 1
and 2 or positions 3 and 4.)）
A 1-2-3-4
Position C B 1-2 / 3-4
(first click stop Free (switching possible) C 1-2-3-4
position)
Lever position

15.2 Selecting Filters

A filter cube consists of three types of optical components: an

Barrier filter
excitation filter (EX filter), a barrier filter (BA filter), and a dichroic UV-2A EX 33

mirror (DM). Select the filter cube with the desired combination of DM 400-380
BA 42 0
0

optical components to suit the characteristics of the specimen

and the fluorophore by referencing the properties of each filter.
Excitation filter
You can select a combination of an excitation filter and a barrier
filter even if you are using the same excitation method.
Excitation filters, barrier filters, and dichroic mirrors can be
Dichroic mirror (inside the cube)
purchased separately.
Since excitation filters are exposed to strong light during Filter cube
operations, they will deteriorate over time. Replace the filter at
intervals determined by usage.

Spacer inside the filter cube

Some types of filter cubes cannot be inserted directly into the epi-fluorescence attachment.
Follow the procedure described in Chapter 3, Section 6 “Attaching a filter cube” to remove an internal spacer or
reverse the spacer before insertion.
Chapter 2
Individual Operations

Excitation filter (EX filter)

Excitation filters allow selective transmission of light (excitation

light) in the wavelength range required for fluorescent light
EX filter
emissions from the specimen, blocking light of all other
Bandwidth
transmission

wavelengths. The range of wavelengths allowed to pass through

a filter is referred to as the bandwidth.
Spectral

The bandwidth range of an excitation filter determines the

brightness of the fluorescent image, the generation of
autofluorescence (fluorescence resulting from substances other
than the fluorophores), and degree of fading. The broader the
bandwidth, the greater the amount of excitation light irradiated 0 Wavelength
onto the specimen, thereby increasing the brightness. However,
this also increases the amount of autofluorescence and causes EX filter bandwidth
faster color fading. Narrow bandwidth reduces the amount of
excitation light striking the specimen and causes the image to
appear darker, but reduces autofluorescence and fading. For
specimens with pronounced autofluorescence, use excitation
filters with a narrow bandwidth. (Note that this will make the
fluorescent image darker.)
Since excitation filters are exposed to strong light during
operations, they will deteriorate over time. Replace the filter at
intervals determined by usage.

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 Chapter 2 Individual Operations

EX Filter Bandwidth and Fluorescent Image

Narrow EX filter bandwidth Wide

Brightness of Dark Bright

fluorescent image

Generation of Low High

autofluorescence

Degree of color fading Low High

Barrier filter (BA filter)

Barrier filters allow only fluorescent light emitted by the specimen to pass, blocking excitation light. This allows the
fluorescent image to be viewed without excess illumination (dark background).
There are two types of barrier filters: LP filters that block all light below a certain wavelength but pass all light of longer
wavelengths, and BP filters that pass light of a certain waveband and block all other light. Use the filter type appropriate
for your intended purpose.

LP filter (long-pass filter)

LP filters block all light below a certain wavelength but pass all
light of longer wavelengths. The border wavelength is called the
cut-on wavelength.
(1) For specimens labeled with a fluorophore in which the
fluorescent waveband and excitation waveband (light that the
specimen absorbs in order to emit fluorescent light) are very
close, selecting a barrier filter with the shortest cut-on
wavelength permitted by the performance requirements will
result in most efficient fluorescent microscopy. If the cut-on
LP520
wavelength is long, excitation light and fluorescent light will
be entirely distinct, tending to darken the background of Waveband
transmission

fluorescent images. However, recent developments in filter emitted by Waveband

FITC
Spectral

performance have resulted in increased use of filters of short emitted by

TRITC
cut-on wavelengths.
(2) For multiple-labeled specimens, use an LP filter for
Chapter 2
microscopy of fluorescent images of all fluorophores. Note
that a combination involving an ordinary dichroic mirror, an Wavelength

Individual Operations
excitation filter, and an LP-filter-type barrier filter will be
Long-pass filter
incapable of excited fluorophores that emit long-wavelength
fluorescent light – for example, TRITC in the case of FITC (Both FITC and TRITC images are visible.)
and TRITC. This will result in very dark TRITC fluorescent
images. For such cases, we recommend using multiband
filters.

BP filter (bandpass filter)

Bandpass filters pass only light of a certain wavelength range, BA520-560 (BP type)
blocking all other light.
transmission

BP filters are used for microscopy of fluorescent images involving

Waveband Waveband
Spectral

a specific fluorophore in multiple-labeled specimens. (For emitted by emitted by

example, in a double-labeled specimen of FITC and TRITC, the FITC TRITC
BA520-560 filter enables microscopy of just the FITC fluorescent
image.)
However, BP filters will not separate autofluorescence, if any Wavelength
(because the fluorescent image in the above combination is
green only). LP filters are better suited for making fine separation Band-pass filter
of autofluorescence based on slight color differences. (Only the FITC fluorescent image is visible.)

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 Chapter 2 Individual Operations

16 Capturing Images

When the DS-U3 or DS-L3 DS Camera Control Unit is connected

to the microscope, you can use the Capture button on the base of Camera head
the microscope to capture digital images with ease.
When using a tube that allows for light distribution to the
binocular section and the camera port, images can also be
captured while in an observation posture from the binocular
section.

Capture

TOR
QUE
button

MP
CLA
ND4 ND8
OUT

IN

DS camera
control unit
(DS-L3 in the figure)

Photomicroscopy

16.1 Photomicroscopy

The photomicroscopy procedure is described below. Also refer to the instruction manual provided with the DS-U3, DS-L3,
or the camera's control software for the details including the camera settings.
In addition, when using the DS-L3, you must configure at least the following information:
• Folder for data storage. Camera cable
• File name for saving the file connector
• File format and file size Camera head
• Date and destination of data
(1) Adjust the illumination of the microscope correctly, and C-mount
adjust the focus onto the specimen image.
Camera fine
(2) Adjust the installation position of the camera head focus adjustment
based on the stage movement direction. ring
Chapter 2 Camera
Loosen the attachment guide fixing screw on the C mount,
centering screws
and adjust the camera position and rotation. The movement
Attachment guide (two)
Individual Operations

on the monitor should be in the opposite direction of the fixing screw

stage. (When the stage is moved in the direction from left to
right, the image on the monitor should move from right to Adjusting the camera head
left.) After making the appropriate adjustments, tighten the attachment position
screws firmly.
(3) Focus the image.
If the image viewed through the eyepiece appears to be in
focus but the image on the monitor is out of focus, turn the
camera fine focus adjustment ring on the C mount until the
image on the monitor is in focus. Note that such out of focus
situations may also indicate incorrect diopter adjustment.
Make sure you have made diopter adjustments as well.
(→Chapter 2 “4 Adjusting the Diopter”)
(4) Center the camera.
Turn the right and left camera centering screws to align the image through the eyepiece with the image on the
monitor.
(5) Select the camera scene mode suitable for the microscopy method.
(6) Adjust the camera’s white balance.
To adjust the white balance, press the WB button while capturing an image of a clear section of the specimen slide.
(For fluorescent photomicrography, it is recommended that you adjust the white balance under normal bright-field
microscopy conditions before capturing images.)

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 Chapter 2 Individual Operations

NCB filter (for Ci-S only)

Ci-S has a built-in NCB filter for the color temperature conversion.

(7) Align the specimen.

(8) Readjust the focus onto the target.
(9) Adjust the image brightness using the camera exposure compensation function.
(10) Check the image using the Freeze button.

(11) If the image is acceptable, press the Capture button to save the image.
(The operating procedure differs if the DF/FL scene mode is selected. For details, refer to the operating manual provided
with the camera.)

Number of images captured

Seriography is not available because the Capture button is designed to capture only one image.

16.2 Tips on Microscope Settings for Photomicroscopy

Adjusting the light intensity

Lamp/LED: When Ci-S is used in applications for which accurate color reproduction is critical, turn the brightness control
knob to the mark and use ND filters for brightness adjustments.
Filter: Place a commercially available color compensation filter on the field lens on the the microscope base, inside
or on the filter cassette holder as necessary.

Adjusting the condenser

• Always focus and center the condenser.

• For phase contrast microscopy, center the annular diaphragm.
• The diaphragm aperture should generally be adjusted to 70 to 80% of the numerical aperture of the objective.

Confirming the photomicrographic range Chapter 2

The image on the monitor represents the photomicrographic range.

Individual Operations
Confirming the focus

Check the focus both through the eyepiece and on the monitor. If the focal positions for the two images differ, adjust the
camera fine focus adjustment ring at the camera port.

Adjusting to keep out extraneous light

Field diaphragm: Stop down the diaphragm to a setting just slightly wider than the area shown on the monitor.
Eyepiece: Cover the eyepiece with a piece of cloth or similar.

Protecting fluorescent images from decoloration

The fluorescence of specimens may fade during exposure. To prevent this, do the following:

■ Use a brighter optical system combination

Even if the overall magnification is the same on the monitor, the combination of objective and camera zoom can
result in significant variations in exposure time. Nikon recommends increasing the magnification of the objective
rather than of the zoom. (Generally, the numerical aperture of the objective increases with magnification. The larger
the numerical aperture, the brighter the resulting image.)

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 Chapter 2 Individual Operations

■ Adjusting the excitation light

Excessively bright excitation light will accelerate the decoloration of the specimen, making it more difficult to acquire
suitable fluorescent images. Insert ND filters into the optical path to adjust the brightness.

■ Specimen
Photomicrography of faded specimen sections requires prolonged exposure time and results in poor color
reproduction and low-quality images. Move the specimen to obtain images from a fresh section of the specimen
previously unexposed to excitation light. For best results, use the phase contrast method to select a specimen
section for photomicrography, and then switch to the fluorescent method to capture images.

Enhancing the contrast of a simple or sensitive tint plate image

With Ci-S, since the infrared light from the illuminator reduces the
contrast of an image, insert an optional IR cut filter in the field lens
C-SP Polarizer Unit
before attaching the polarizer unit.
for Simple
Cover the IR cut filter for the simple polarizer over the field lens. Polarization

Screw the IR cut filter for a sensitive tint plate polarizer into the
bottom of the polarizer unit for first-order red compensation. [2]
IR cut
filter for C-SP OUT
ND4 ND8

[1] IN

Field lens

C-TP Polarizer Unit for

First-order Red
Compensation

[1]
IR cut
filter for C-TP
ND4 ND8

[2]
OUT

Chapter 2 IN
Individual Operations

Field lens

Attaching an infrared ray blocking filter

Adjusting the brightness of the image on the monitor

When observing images captured by the camera and displayed on the monitor, you can adjust the brightness by varying
camera adjustment parameters, such as display mode, exposure mode, photometry mode, exposure compensation, and
image level adjustment.
See the instruction manual provided with the DS-U3, DS-L3, or the camera's control software for details.

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 3 Assembly

1 ECLIPSE Ci-S/Ci-L System Configuration

ENG mount camera

C mount camera
photomicrography
device

TV adapter

Centering telescope CFI eyepiece

Eyelevel riser

Camera head
Analyzer unit

Binocular tube Trinocular tube Ergonomic tube DSC port

Filter cube

CI-FL
epi-fluorescence
attachment
Spacer for Camera
the nosepiece
cable

Analyzer Sextuple
slider Sextuple Ci-S/Ci-L main body
nosepiece nosepiece
with DIGITAL SIGHT CAPTURE

analyzer slot
DS-U3/L3 DS
camera control unit
Camera
trigger cable
CFI objective

HG precentered
fiber illuminator

Achr-Apl N.A=1.4
Chapter 3
1.4 1.2 1.0 0.8 0.6 0.4 0.2

Stage

Assembly
Condenser

Filter Polarizer
cassette holder unit

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 Chapter 3 Assembly

2 Assembly for Bright-field Microscopy

[Tool for assembly: hex driver]

1 Checking the input voltage

Check the input voltage indicated on the back of the
microscope. Use the microscope only if the indicated
input voltage matches the power supply voltage for the
area in which the microscope will be used.
NIKON
CORPOR
TOKYO,
JAPAN ATION MODE

Input voltage
100–240 L ECLIP
0.9A
V~ SE Ci-S
50/60Hz
9400
This
device MADE 01
Rules. complies IN CHINA
Operation with Part
conditions is subject 15 of
the FCC
(1) this : to the
4N75 device following
INSPECT and (2) may not two
this cause
EQUIPMEION received, device
must harmful
NT undesired including accept interferen
interferen any interferen ce,

label
operation ce that
This
Class
. may causece
Canadian A digital
apparatus
Cet appareilICES-003 complies
.
confirme numériqu with
à la norme e de la
classe
NMB-003 A est
du Canada.

WARNING 80
90
0

10
If the indicated voltage and the supplied voltage

70

20
60
30

50
40

differ, do not attempt to use the microscope. Contact DSC

your nearest Nikon representative for advice.

Checking the input voltage

2 Attaching the stage

Stage clamp screw
(1) Turn the coarse focus knob to remove the
cushioning material from the substage section.
(2) Turn the coarse focus knob until the elevating
section is brought to the lowermost position.
(3) Place the stage on the substage and fix into place
with the stage clamp screw.
TO RQ
UE

P
CL AM
ND4 ND8
OUT

IN

Attaching the stage

Using the filter cassette holder

Insert the filter cassette holder on the field lens of
Chapter 3 the microscope after the stage has been raised up
to the upper limit.
TO RQ
Assembly

UE

P
CL AM

ND4 ND8
OUT

IN

Filter cassette holder

Attaching a filter cassette holder

Exclusivity of the filter cassette holder

and the spacer for the nosepiece
The filter cassette holder cannot be used
simultaneously with the spacer for the nosepiece.

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 Chapter 3 Assembly

3 Attaching the condenser

(1) Turn the coarse focus knob until the elevating
Condenser focus knob
section is brought to the upper limit.
(2) Turn the condenser focus knob until the condenser
holder reaches the bottom.
(3) Insert the condenser, adjust it to face the front, then
secure it in place with the tool.
(4) Turn the condenser focus knob until the condenser

P
CL AM
80

holder reaches the top.

TO RQ
70 90

UE
60

0
50

10
40

20
30

POWER

Condenser clamp screw

(use a tool)

Attaching the condenser

4 Attaching the tube

Place the tube on the microscope arm and secure it by
tightening the clamp screw on the arm.

Tube clamp
screw

Attaching the tube

Using the eyelevel riser

Place the eyelevel riser on the microscope arm and
secure it by tightening the clamp screw on the arm.
Attach the tube on the eyelevel riser and secure it
by tightening the clamp screw on the eyelevel riser Tube clamp screw
using a tool. (use a tool)
Chapter 3

Eyelevel riser Assembly

Eyelevel riser
clamp screw

Attaching the eyelevel riser

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 Chapter 3 Assembly

5 Attaching eyepieces
Make sure the notch on the eyepiece side and the Protrusion Notch

protrusion of the eyepiece sleeve are aligned, then insert

the eyepieces.

Notch on eyepiece
Eyepiece has a notch to prevent rotation. When
attaching, match the notch with the protrusion on
the eyepiece sleeve. Otherwise, eyepiece is not
attached to the correct position.
Attaching eyepieces

6 Attaching the nosepiece

Place the nosepiece under the fitting part, at a position
slightly toward yourself. (Continue sliding the nosepiece
until its front position is aligned with that of the fitting
part.) Secure it in place with the tool provided with the
microscope. Nosepiece clamp screw
(use a tool)

Attaching the nosepiece

Using the spacer for the nosepiece

Follow the procedure below to attach the spacer for
the nosepiece.
(1) Slide the spacer for the nosepiece along the Spacer for
groove to attach it to the nosepiece, and the nosepiece
secure it in place with the provided tool. clamp screw
(use a tool)
(2) Lower the stage to the limit.
Spacer for
(3) Place the nosepiece with the spacer on the
the nosepiece
microscope following the procedure described
in “Attaching the nosepiece” on the previous Nosepiece
page. clamp screw
Chapter 3
(use a tool)
Attaching the spacer for the
Assembly

nosepiece

Exclusivity of the spacer for the nosepiece and the filter cassette holder
The spacer for the nosepiece cannot be used simultaneously with the filter cassette holder placed on the field
lens.

7 Attaching the objective

Screw the objective into the nosepiece. When attaching the objective in this way, make sure that the magnification
of the objective increases when the nosepiece is turned clockwise (clockwise when viewed from above the
eyepiece).

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 Chapter 3 Assembly

8 Connecting the power cord

WARNING
Be sure to use the specified power cord. Use of other power cords may result in malfunction or fire. This
product is classified as having Class I protection against electric shock. Make sure this product is connected
to an appropriate protective earth terminal.
Refer to Chapter 6, “2 Performance Properties” for the specified power cords.
To prevent electric shock, always turn off the power switch (press to the “O” position) for the microscope
before connecting or disconnecting the power cord.

(1) Check that the microscope power switch is OFF. Power switch OFF
(press to “O” position)
(2) Plug the power cord into the AC inlet at the back of ND4 ND
OUT
8

the microscope. IN

(3) Plug the other end of the power cord into a wall
outlet.

AC inlet

Assembly of the system configuration required for bright-field DS C

microscopy is now completed.

Connecting the Power Cord

Chapter 3

Assembly

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 Chapter 3 Assembly

3 Assembly for Phase Contrast Microscopy

Follow the procedure described in Section 2 “Assembly for Bright-field Microscopy” to perform assembly.
Note the following:

■ Attaching a condenser for phase contrast microscopy

Use a condenser for the phase contrast microscopy in the step 3, “Attaching the condenser.”
The procedure is the same.

■ Attaching a Ph objective
A Ph objective must be attached in the step 7 “Attaching the objective”.
The procedure is the same.

4 Assembly for the Simple Polarizing Microscopy

Follow the procedure described in Section 2 “Assembly for Bright-field Microscopy” to perform assembly.
Note the following:

■ Attaching an analyzer tube for simple polarization

Attach an analyzer tube for simple polarization to the

microscope arm prior to the step 4, “Attaching the tube.”
Analyzer
Place the analyzer unit (analyzer tube for simple IN/OUT knob
polarization) on the arm so that the analyzer IN/OUT knob Tube clamp
is positioned to the right, and secure the analyzer unit in screw
place with the analyzer unit clamp screw. (use a tool)

Place the tube on the analyzer unit and tighten the tube
Analyzer tube for
clamp screws of the analyzer unit using the tool provided simple polarization
with the microscope to secure the tube.

Slider-type analyzer for simple polarization Analyzer unit

clamp screw
When using the D-SA Analyzer Slider for Simple
Polarization instead of an analyzer tube for simple
TO RQ

polarization, insert it into the analyzer slot of the nosepiece.

UE

P
CL AM

(To use the D-SA Analyzer Slider for Simple Polarization, OUT
ND4 ND8

Chapter 3 the C-NA sextuple nosepiece with analyzer slot is IN

required.)
Assembly

Polarizer unit
Polarizer unit for clamp screw (use a tool)
simple polarization

Attaching an analyzer tube for simple polarization,

polarizer unit for simple polarization

■ Attaching a polarizer unit for simple polarization

Set the polarizer unit for simple polarization over the field lens on the base of the microscope. Make sure the
orientation mark on the polarizer comes to the front at this point and tighten the polarizer unit clamp screw.
The actual securing will take place after the orientation of the analyzer and polarizer has been adjusted in Step 13 of
the simple polarizing microscopy procedure.

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 Chapter 3 Assembly

5 Assembly for Sensitive Polarization Microscopy

Follow the procedure described in Section 2 “Assembly for Bright-field Microscopy” to perform assembly.
Note the following:

■ Attaching an analyzer tube for first-order red compensation

Attach an analyzer tube for first-order red compensation to

the microscope arm prior to the step 4, “Attaching the tube.”
Place the analyzer unit (analyzer tube for first-order red Analyzer
Tube clamp screw IN/OUT knob
compensation) on the arm so that the analyzer IN/OUT (use a tool)
knob is positioned to the right, and secure the analyzer unit
in place with the analyzer unit clamp screw. Analyzer tube for
first-order
Place the tube on the analyzer unit and tighten the tube red compensation
clamp screws of the analyzer unit using the tool provided
with the microscope to secure the tube. Analyzer unit
clamp screw

Slider-type analyzer for first-order red

compensation
When using the C-AS Analyzer Slider for First-order Red Polarizer unit for

TO RQ
UE
first-order
Compensation instead of an analyzer tube for first-order

P
CL AM
red compensation
ND4 ND8
OUT

red compensation, insert it into the analyzer slot of the IN

nosepiece. (To use the C-AS Analyzer Slider for First-order

Red Compensation, the C-NA sextuple nosepiece with
analyzer slot is required.) Polarizer unit
clamp screw (use a tool)

Attaching an analyzer tube for first-order red

compensation,
polarizer unit for first-order red compensation

■ Attaching a polarizer unit for first-order red compensation

Set the polarizer unit for first-order red compensation over the field lens on the base of the microscope. Make sure
the orientation mark on the polarizer comes to the front at this point and tighten the polarizer unit clamp screw.
The actual securing will take place after the orientation of the analyzer and polarizer has been adjusted in Step 13 of
the sensitive polarization microscopy procedure.

Chapter 3

Assembly

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 Chapter 3 Assembly

6 Assembly for Epi-fluorescence Microscopy

Follow the procedure described in Section 2 “Assembly for Bright-field Microscopy” to perform assembly.
Note the following:

■ Attaching the epi-fluorescence attachment

Attach the epi-fluorescence attachment to the microscope arm prior to the step 4, “Attaching the tube.”

(1) Place the epi-fluorescence attachment on the Epi-fl attachment clamp bolt
microscope arm. (use a tool)

(2) Secure it by tightening the clamp screw on the arm and

HG adapter
two fixing bolts at the rear of the epi-fluorescence
attachment. Use a hex wrench provided with the
Tube clamp
epi-fluorescence attachment to tighten the fixing bolts. screw
(3) Attach the HG adapter of the mercury lamp illuminator (use a tool) A
1-2-
B 3-4
1-2 /
C 3-4

4
1-2-
3-4

to be used to the bayonet mount on the back. (For CUBE

1

detailed information, refer to the instruction manual

1 2 3 4

provided with the mercury lamp illuminator.)

(4) Place the tube on the epi-fluorescence attachment and
Epi-fluorescence Epi-fluorescence
tighten the tube clamp screws of the epi-fluorescence attachment attachment
attachment using the tool provided with the microscope clamp screw
to secure the tube. Attaching the epi-fluorescence attachment

■ Attaching a filter cube

WARNING
Always turn off the mercury lamp before inserting or removing a filter cube.

(1) Remove the filter cube replacement cover on the left of

the epi-fluorescence attachment.
(2) Insert a filter cube.
ND4

The filter cubes listed below cannot be inserted directly

ND8
ND16
3-4
1-2-
A 3-4
1-2 /
B 3-4
1-2-
C

into the filter bay of the epi-fluorescence attachment.

Remove an internal spacer or reverse the spacer
before insertion.
Chapter 3 <Filter cubes requiring spacer removal> Filter cube
replacement cover
• UV-2A
Assembly

• UV-2B

<Filter cubes requiring spacer reversal> Filter cube

A

3-4
1-2-
A 3-4

• DAPI
1-2 /
B 3-4
1-2-
C

• FITC
UV-2A EX 330-380
DM 400
BA 420

• GFP-L
• GFP-B
• TRITC
• Tx-Red Attaching a filter cube

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 Chapter 3 Assembly

●Removing/attaching the internal spacer● Filter cube

1) Place the filter cube on a work table with the
excitation filter facing up.
2) Unscrew the ring retaining the excitation filter.
(Be careful to avoid dropping the filter.)
3) Remove the spacer inside the removed ring.
4) Remove or reverse the spacer as appropriate for
the particular filter cube type before its insertion.
5) Reattach the ring.

Spacer Ring

Attaching/removing the internal spacer

(3) Insert a nameplate into the position with the same address as the one indicated on the filter cube switching knob
on the right side of the microscope.
(4) Turn the filter cube switching knob and insert other filter cubes into the remaining open bays.
(5) Restore the slot cover back to its original position.

■ Replacing excitation and barrier filters

Turn approx. 30 degrees to
The excitation filter, barrier filter, and dichroic mirror can be secure in place.
removed from the filter cube for replacement.
Excitation filters are screw-in filters, while barrier filters are Barrier filter
slide-in filters.
Align the projection on the barrier filter with the groove on
the filter cube and turn clockwise by approximately 30 Excitation filter
degrees to secure it in place.

Replacing the excitation and barrier filters

■ Attaching a light shielding plate

Secure a light shielding plate in place with the fixing screws

on the lower front of the epi-fluorescence attachment. To Light shielding plate
remove the plate, loosen the fixing screws and pull it clamp screws
forward. (on the back of the
plate) Chapter 3
A

Assembly
1-2-
B 3-4
1-2 /
C 3-4
4

1-2-
3-4

CUBE
1

1 2 3 4

Attaching a light shielding plate

■ Attaching a shielding tube

Attach a shielding tube using the same procedure for attaching a condenser to the condenser holder.

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 Chapter 3 Assembly

7 Attaching a Camera

■ When attaching a camera head to the ergonomic tube

(1) Screw the camera head into the C mount on the DSC
port.
Camera head
(2) Remove the rear cover of the ergonomic tube and
insert the DSC port. C-mount

(3) Secure the DSC port in place with the tool provided
DSC port
with the microscope.
(4) Connect the camera head to the DS-U3 or DS-L3 DS
DSC port
Camera Control Unit with a camera cable. Camera attached in place fixing screw
(5) Use a camera trigger cable to connect the DSC
connector on the back of the microscope to DS-U3 or Attaching a camera head
DS-L3.

Notes when connecting cables

When connecting a capture cable to the DSC connector,
insert it to the end.

Prior to photomicrography, adjust the camera position as

appropriate. (See Chapter 2, “16.1 Photomicroscopy”.)

■ Installing a camera head to the trinocular tube

Attach a C mount camera, ENG mount camera and a photomicrography device via an adapter to the trinocular tube.

Chapter 3
Assembly

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 4 Troubleshooting
Misuse of this product may adversely affect performance, even if this product is properly functional. If any of the following
problems occurs, be sure to check the following table for possible causes before requesting service.
If you detect problems that are not listed below or the problem still persists after measures are taken, turn off the device
and contact your nearest Nikon representative.

1 Optical System and Operation

1.1 General

Problem Cause Measure

Dirt or dust rotates when eyepiece is
turned.
Eyepiece is dirty. Clean the eyepiece.
(→Chapter 5 “2.1 Lens Cleaning”)

Dirt or dust does not rotate when eyepiece

is turned (1) to (5)
(1) The specimen is dirty if dirt or dust (1) Clean the specimen.
moves when specimen is moved on
stage.
(2) The tip of the condenser lens is dirty if (2) Clean the condenser.
Dirty or dusty field of dirt or dust goes in and out of view (→Chapter 5 “2.1 Lens Cleaning”)
view when looking into when the condenser is moved up and
eyepiece. down while using low magnification
objective.
(3) Objective is dirty if dirt or dust (3) Clean the objective.
disappears when the objective is (→Chapter 5 “2.1 Lens Cleaning”)
switched.
(4) Field diaphragm image is not focused (4) Make sure the condenser is focused and
on the specimen surface. (Condenser centered.
is not correctly adjusted.) (→Chapter 2, “5 Focusing and Centering the
Condenser”.)

(5) An aperture diaphragm is stopped (5) Open it to proper size.

down too far. (→Chapter 2 “6 Adjusting the Aperture Diaphragm”)

Dirt or dust on the monitor moves when

camera is turned
Lenses or specimen are dirty or Check and clean by following the procedure in
dusty. “If dirt or dust does not turn by rotating the
Dirt or dust is displayed eyepiece” in “When looking into the eyepiece,
on the monitor. dirty or dusty field of view”.
Chapter 4
Dirt or dust on the monitor does not move
when camera is turned
Troubleshooting

Camera is dirty. Detach the camera and clean it by following

the instruction manual of the camera.
No cover glass is attached. Attach a cover glass of the specified thickness
The thickness of the cover glass is (0.17 mm). (However, no cover glass is
Image quality is poor. required for an NCG objective.)
inadequate.
Contrast is poor.
Resolution is poor. Objective correction ring does not match
the thickness of the cover glass. (for the Correct the ring as appropriate.
objective with a correction ring)

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 Chapter 4 Troubleshooting

Problem Cause Measure

Check and clean by following the procedure in
“If dirt or dust does not turn by rotating the
Lens and specimen are dirty or dusty.
eyepiece” in “When looking into the eyepiece,
dirty or dusty field of view”.
An aperture diaphragm is stopped down Open it to proper size.
too far. Otherwise, it is open too much. (→Chapter 2 “6 Adjusting the Aperture Diaphragm”)

Make sure the condenser is focused and

Field diaphragm image is not focused on centered.
Image quality is poor. the specimen surface.
(→Chapter 2, “5 Focusing and Centering the
Contrast is poor. (Condenser is not correctly adjusted.)
Condenser”.)
Resolution is poor.
No immersion oil is applied to the tip of an Apply our designated non-fluorescent
oil-immersion objective. immersion oil.
The designated immersion oil is not used. (→Chapter 2 “12 Oil Immersion”)

Remove the air bubbles.

The immersion oil contains air bubbles.
(→Chapter 2 “12 Oil Immersion”)

Immersion oil adhering to the tip of the Clean it as appropriate.

dry-type objective. (→Chapter 2 “12 Oil Immersion”)

Place ND filters in the optical path.

ND filters are out of the optical path.
(→Chapter 2 “1.2 Adjustment with ND Filters”)

Field of view is too Turn the brightness control knob to the

bright. mark position and adjust the brightness with
The lamp voltage is too high. ND filters.
(→Chapter 2, “1 Adjusting the Brightness of a
Diascopic Image”)

Turn the brightness control knob to the

mark position and adjust the brightness with
Lamp voltage is too low. ND filters.
(→Chapter 2, “1 Adjusting the Brightness of a
Diascopic Image”)

This should normally be adjusted to 70 to 80%

Condenser aperture diaphragm is stopped of numerical aperture of the objective.
down too far.
(→Chapter 2 “6 Adjusting the Aperture Diaphragm”)

Field of view is too dark. Make sure the condenser is focused and
Field diaphragm image is not focused on centered.
the specimen surface. (→Chapter 2, “5 Focusing and Centering the
Condenser”.)

Set to binocular 100%.

Optical path is not switched to binocular
(→Chapter 2, “9 Switching the Optical Path of the
100%.
Tube”)

Chapter 4 The lamp has reached the end of its Replace the lamp.
product life (for Ci-S). (→Chapter 5 “1 Replacing the Lamp (for Ci-S)”)
Troubleshooting

Turn the brightness control knob to the

mark position and adjust the brightness with
Image is yellowish or Lamp voltage is too low or too high. (for ND filters.
very bluish. Ci-S)
(→Chapter 2, “1 Adjusting the Brightness of a
Diascopic Image”)

Color of the image

visible to the naked eye
White balance of the camera is not set Set the white balance according to camera's
is different from that of
properly. instruction manual.
the image displayed on
the monitor.

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 Chapter 4 Troubleshooting

Problem Cause Measure

The entire field of view is Filter cube is in the optical path when non
Remove the filter cube from the optical path.
bluish or yellowish. epi-fluorescence observation.
Confirm that parts (nosepiece, condenser, etc.)
are correctly attached.
Parts are attached incorrectly.
(→Chapter 3, “2 Assembly for Bright-field
Lack of visibility around Microscopy”)
periphery of field of view. Correctly set the optical path switching lever,
Illumination is uneven nosepiece, filter cube switchover turret,
Movable parts are not seated correctly.
across the field of view. condenser turret, and slider, etc. (Move the
Field of view is not part until it clicks.)
visible. Make sure the condenser is focused and
Field diaphragm image is not focused on centered.
the specimen surface. (→Chapter 2, “5 Focusing and Centering the
Condenser”.)

Open the field diaphragm slightly wider than

Field diaphragm is stopped down too far. the field of view.
(→Chapter 2 “8 Adjusting the Field Diaphragm”)

Lack of visibility around Incorrect combination of the objective with Adopt an appropriate combination.
periphery of field of view. the condenser. (→Chapter 2 “7 Selecting a Condenser”)
Illumination is uneven
In low magnification microscopy, required Perform the operation such as swinging out or
across the field of view.
condenser operation was not performed. sliding the condenser.
Field of view is not
visible. Attach it correctly.
A lamp is attached incorrectly.
(→Chapter 5 “1 Replacing the Lamp (for Ci-S)”)

Clean them as appropriate.

Lens and specimen are dirty or dusty.
(→Chapter 5 “2.1 Lense Cleaning”)

Turn up the cover glass and attach it to the

stage.
The specimen is upside down. (→Chapter 2 “2.1 Bright-field Microscopy Procedure
- 5 Place a specimen on the stage, and move the
stage to bring the target into view”)

Out of focus with an The thickness of the cover glass is Attach a cover glass of the specified thickness
objective of high inadequate. (0.17 mm).
magnification. Some objective has a stopper to keep the
pushed in state. Turn the tip of the object to
release.
Fail-safe device for specimen damage
Tip of the objectives without stopper can not
protection of the objective is pushed in.
be rotated. Do not use force to draw the
stopper. Contact your nearest Nikon
representative.
Screw the objective all the way in.
A focal deviation is high An objective is attached incorrectly. (→Chapter 3, “2 Assembly for Bright-field
Chapter 4
when switching over Microscopy ― 7 Attaching the objective”)
objectives.
Troubleshooting

Diopter adjustment has not been Perform diopter adjustment.

performed. (→Chapter 2 “4 Adjusting the Diopter”)

Attach it correctly.
Image is not in focus The stage is attached incorrectly. (→Chapter 3, “2 Assembly for Bright-field
although the stage is Microscopy ― 2 Attaching the stage”)
raised to the highest
position. Refocusing position is set lower than the Check and reset the settings.
focusing position. (→Chapter 2 “2.4 Refocusing”)

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 Chapter 4 Troubleshooting

Problem Cause Measure

Attach it correctly and rotate to the click stop
The nosepiece is not attached correctly, or position.
has not been fully rotated to the click stop
(→Chapter 3, “2 Assembly for Bright-field
position.
Microscopy ― 6 Attaching the nosepiece”)

One side of the field of Position the specimen in place on the stage.
view (up, down, right, or The specimen is tilted relative to the stage (→Chapter 2 “2.1 Bright-field Microscopy Procedure
left) is not focused. surface. - 5 Place a specimen on the stage, and move the
The image flows (i.e. stage to bring the target into view”)
becomes asymmetrically
defocused when moving Attach the stage correctly.
the focal point) The stage is tilted. (→Chapter 3, “2 Assembly for Bright-field
Microscopy ― 2 Attaching the stage”)

Attach the condenser correctly.

The condenser is tilted. (→Chapter 3, “2 Assembly for Bright-field
Microscopy ― 3 Attaching the condenser”)

Perform interpupillary adjustment.

Interpupillary adjustment has not been
(→Chapter 2 “2.1 Bright-Field Microscopy
Images in left and right performed.
Procedure ― 8 Adjust the interpupillary distance”)
eyepieces are not
coincident. Perform diopter adjustment.
Diopter adjustment has not been
(→See Chapter 2 “4 Adjusting the Diopter” in the
performed.
“Operation”.)

Perform diopter adjustment.

Diopter adjustment has not been
(→See Chapter 2 “4 Adjusting the Diopter” in the
performed.
“Operation”.)

Eyes become fatigued. Adjust the brightness using the brightness

control knob or ND filters to attain a suitable
Brightness is inadequate. brightness.
(→Chapter 2, “1 Adjusting the Brightness of a
Diascopic Image”)

Fix the holder securely.

The specimen holder is not securely-fixed (→Chapter 2 “2.1 Bright-field Microscopy Procedure
on the stage. - 5 Place a specimen on the stage, and move the
The specimen does not stage to bring the target into view”)
move smoothly.
Adjust to appropriate torque weight.
Rotating torque of the stage knob is set too
(→Chapter 3 “3.3 Adjusting the Knob Rotation
heavy.
Torque”)

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Troubleshooting

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 Chapter 4 Troubleshooting

1.2 Epi-fluorescence Microscopy

Problem Cause Measure

Lack of visibility around
periphery of field of view.
Push the cube in to the limit.
Illumination is uneven
The filter cube is misaligned. (→Chapter 3, “6 Assembly for Epi-fluorescence
across the field of view.
Microscopy ― ■ Attaching a filter cube”)
Field of view is not
visible.
Open the shutter.
(→Chapter 2, “15 Tips for Epi-fluorescence
A fluorescent image is The shutter is closed. Microscopy ― Protecting the specimen and
not visible (when the preventing it from decoloration (shutter for the
lamp is ON). epi-fluorescence attachment”)

Use a correct filter cube.

The selection of the filter cube is incorrect.
(→Chapter 2, “15.2 Selecting Filters”)

Remove the ND filters from the optical path as

necessary.
ND filters of the epi-fluorescence (→Chapter 2, “15 Tips for Epi-fluorescence
attachment are in the optical path. Microscopy ― Adjusting the brightness of the
fluorescent image (adjusting the ND) ― ■ ND filters
in the epi-fluorescence attachment”)

Light intensity is set to too low in the

Adjust it.
setting of the ND on the HG precentered
(→Check your illuminator's manual.)
fiber illuminator.
Change the light source to a mercury lamp.
A halogen light source is used for a dark (→Chapter 2, “15 Tips for Epi-fluorescence
The fluorescent image is Microscopy ― Protecting the specimen and
very dark (when the specimen.
preventing it from decoloration (shutter for the
lamp is ON). epi-fluorescence attachment”)
Mercury lamp on the HG precentered fiber Replace the lamp.
illuminator has reached the end of its
(→Check your illuminator's manual.)
product life.
A designated objective is not used at UV or
Use a designated objective.
V excitation.
The room is bright. Make it darker.

The optical path switching lever is not set Switch the lever position to 100% light
distribution for the binocular section.
to 100% light distribution for the binocular
(→Chapter 2, “9 Switching the Optical Path of the
section. Tube”)

The dia-illumination lamp is on. Turn off the dia-illumination lamp.

The fluorescent image
quality is poor. The filter cube being used is not suitable Use a filter cube suitable for the specimen.
for the specimen. (→Chapter 2, “15.2 Selecting Filters”)
Chapter 4
Clean it as appropriate.
The objective or cover glass is dirty.
Troubleshooting

(→Chapter 5 “2.1 Lense Cleaning”)

Use our designated non-fluorescent

immersion oil.
The immersion oil is fluorescent.
The contrast of the (→Chapter 2, “15 Tips for Epi-fluorescence
fluorescent image is Microscopy”)
poor. Use a non-fluorescent slide.
The slide is fluorescent. (→Chapter 2, “15 Tips for Epi-fluorescence
Microscopy”)

Lower the condenser, or remove the

Stray light is entering from the condenser.
condenser and attach a shielding tube.

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 Chapter 4 Troubleshooting

1.3 Phase Contrast Microscopy

Problem Cause Measure

The Ph annular diaphragm of the Adjust so that they match.
condenser does not match the phase plate (→Chapter 1, “2.2 Phase Contrast Microscopy
image of the objective. Procedure ― 12 Center the Ph annular diaphragm”)

Put the Ph annular diaphragm with the same

The Ph annular diaphragm of the Ph code as the objective into the optical path.
condenser and the objective selected do (→Chapter 1, “2.2 Phase Contrast Microscopy
not match. Procedure ― 14 Adjust the Ph annular diaphragm in
Poor contrast. the condenser with the Ph objective to be used.”)

Change the mounting agent or thickness of

The phase contrast of the specimen is too the specimen when preparing the specimen.
large. (→Chapter 2, “14 Tips for Phase Contrast
Microscopy”)

Use a Ph objective suitable for the specimen.

The type of the Ph objective is not suitable
(→Chapter 2, “14 Tips for Phase Contrast
for phase contrast of the specimen.
Microscopy”)

Chapter 4
Troubleshooting

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 Chapter 4 Troubleshooting

2 Electrical requirements

2.1 General

■ Power supply
Problem Cause Measure
There is no power even Connect this product correctly to the network.
The power cord is not connected, or is
though the power switch (→Chapter 3, “2 Assembly for Bright-field
connected improperly.
is on. Microscopy ― 8 Connecting the Power Cord”)

■ Illumination
Problem Cause Measure
Plug in the power cord.
There is no power supplied. (→Chapter 3, “2 Assembly for Bright-field
Microscopy ― 8 Connecting the Power Cord”)

Lamp does not light. Replace the lamp with the specified type.
The lamp has burned out (for Ci-S)
(→Chapter 5 “1 Replacing the Lamp (for Ci-S)”)

Attach a designated lamp.

The lamp is not attached.(for Ci-S)
(→Chapter 5 “1 Replacing the Lamp (for Ci-S)”)

■ Capture button
Problem Cause Measure
The capture button does Camera trigger cable is not properly Attach it correctly.
not function. connected. (→Chapter 3, “7 Attaching a Camera”)

2.2 Epi-fluorescence Microscopy

Problem Cause Measure

Plug in the power cord.
There is no power supplied.
(→Check your illuminator's manual)

Replace the lamp with the specified type.

Lamp has burned out.
The mercury lamp does (→Check your illuminator's manual)
not work. Attach a designated lamp.
The mercury lamp is not attached.
(→Check your illuminator's manual)

The mercury lamp's connector is not Connect it to the illuminator.

connected to the illuminator. (→Check your illuminator's manual)
Chapter 4
Replace the lamp with the specified type.
Troubleshooting

The mercury lamp burns (→Check your illuminator's manual)

The lamp type is incorrect.
out soon after it is turned If the lamp burns out immediately after the
Lamp is at end of its life.
on. replacement, please contact your nearest
Nikon representative.

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Chapter 4
Troubleshooting

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 5 Maintenance and Storage

1 Replacing the Lamp (for Ci-S)

CAUTION
• Beware of burns: Wait until the lamp and nearby parts have cooled before replacing the lamp.
• Beware of electrical shock: Turn off the power switch and unplug the power cord from the wall outlet.
• Beware of abnormal heat generation: Use only the designated lamp.
• Beware of soiling: Avoid touching the glass surface of the lamp with bare hands. Soiling may reduce the service life
of the lamp.
• Lamphouse cover: Make sure the lamphouse cover is securely fitted to the lamphouse after lamp replacement.
• Used lamps: Do not break used lamps. It should be disposed of as industrial waste, according to local regulations
and rules.

(1) Remove the lamphouse cover on the back of the

microscope. Remove the old lamp. Slot
(2) Attach a new lamp. Avoid touching the glass surface of the Cover hook
lamp with your bare hands. Use only the specified lamp
(PHILIPS 5761).
(3) Restore the cover back to its original position. Engage the
cover hook in the slot on the rear of the unit in the opposite
DSC

direction indicated by the arrow in the figure.

Lamp
Lamphouse cover

Replacing the lamp

Chapter 5
Maintenance and Storage

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 Chapter 5 Maintenance and Storage

2 Cleaning

Follow these procedures when cleaning or decontaminating lenses or other parts.

■ Tools used for cleaning

・Blower
・Soft brush
・Soft cotton cloth, lens tissue, or gauze etc.
・Absolute alcohol (ethyl or methyl alcohol), medicinal alcohol
・Petroleum benzine (used only for cleaning immersion oil)

CAUTION
• Petroleum benzine and absolute alcohol used for cleaning are highly flammable. Be careful when handling these
materials, particularly around open flames or when turning the power switch on or off.
• Follow the instructions provided by the manufacturer when using petroleum benzine or absolute alcohol.
• When cleaning the product, do not use organic solvents (alcohol, ether, thinner, etc.) for coated, plastic, or printed
areas. It will result in discoloration or peeling of printed characters.
• Petroleum benzine should be used only to wipe off immersion oil from the objective, and never to clean the entrance
lens at the bottom of the eyepiece tube, prism surface of the eyepiece tube, or the filters.

2.1 Cleaning Lenses

Keep the lens free of dust, fingerprints, etc. If there is contamination on the lenses or filters, image quality decreases. If
any of the lenses
become dirty, clean them by following the procedure given below.

■ Cleaning light dirt (such as dust)

(1) Blow dust off using an air blower.
(2) If this is insufficient, brush away dust with a soft brush or wipe away gently with a piece of gauze.

■ Cleaning tough dirt (such as fingerprint or grease)

Moisten lightly a piece of soft, clean cotton cloth, lens tissue, or gauze with absolute alcohol (ethyl or methyl alcohol)
and wipe the dirt off.

Tips on wiping dirt

Do not reuse cotton cloth, lens tissue, or gauze that have already been used.

2.2 Cleaning Parts Other than Lenses

■ Cleaning light dirt (such as dust)

Wipe with a silicon cloth.

Chapter 5 ■ Cleaning tough dirt (such as fingerprint or grease)

Dampen a piece of gauze with neutral detergent and wipe the dirt gently.
Maintenance and Storage

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 Chapter 5 Maintenance and Storage

2.3 Cleaning the Immersion Oil

(1) Wipe with petroleum benzine.

(2) Finish off the cleaning with absolute alcohol (ethyl or methyl alcohol) after cleaning with petroleum benzine.

If petroleum benzine is not available

If petroleum benzine is unavailable, you may use methyl alcohol. However, typically wipe three or four times
because the detergency is weak.

2.4 Decontaminating the Product

For routine disinfection of this product, Nikon recommends using 70% medical alcohol.
Use of organic solvents on plastic parts may result in discoloration.

Note on disposal
If contact occurs between a sample and this product, determine whether the sample is hazardous. If the sample
is hazardous, follow the standard procedures for your facility.

3 Storage

• Store this product in a dry location where mold is unlikely to form.

Storage conditions are as follows: temperature (-20°C to +60°C), humidity (90% RH max., no condensation)
• Store the objectives and eyepieces in a desiccator or similar container with a drying agent.
• Place a cover over this product to protect it from dust.
• Switch off the microscope (press the switch to the “O” position) and wait for the lamphouse to cool before covering this
product with a cover.

4 Regular Inspections (Charged)

To maintain the performance of this product, Nikon recommends periodic inspections (chargeable service). Contact your
nearest Nikon representative for details.

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 Chapter 5 Maintenance and Storage

Chapter 5
Maintenance and Storage

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 6 Specifications and Safety Standards

1 Microscopy (Operation Principle)

Use objectives and eyepieces of the microscope to magnify minute cells and tissue optically, and manipulate levers and
knobs of the microscope unit to adjust the focus or move the observation point. Then observe or take photographs of the
sample fixed on the slide.

■ Intended use of this product

This microscope is intended for use in microscopic experiment and diagnostics of cells and tissues at hospitals or by
doctors in private practice in the field of pathology, anatomy, and cytology.
The microscopy with diascopic/reflected illuminations is used to observe a sample fixed on the slide (cells and
tissue) as the specimen.
The product is classified as an in-vitro diagnostic medical device.
This product is not intended for use for measurement.
The Z axis position display on the display panel at the front of main body, the XY coordinate position display of
motorized XY stage, and the scale on the stage is an indicator to reproduce the position and does not guarantee the
value of the thickness or length of a sample measured using them.

■ Intended user
It is intended for the medical professional and those who work on experimentations in the field of pathology and
cytology.

2 Performance Properties

■ Nikon Microscope ECLIPSE Ci-S

Model ECLIPSE Ci-S

Optical system Infinity-corrected CF optical system
Objective CFI60
Eyepiece: Field number 22 (with ergonomic tube/binocular tube),
25 (with T/F trinocular tube)
Nosepiece: Sextuple
Focus up/down motion Drive system: Manual coarse/fine motion
(calibration markings for fine motion: 1 µm/marking)
Stroke: 2 mm upward, 28 mm downward
With refocusing mechanism

Specifications and Safety Standards

Light source for 6V30W halogen lamp (PHILIPS 5761)
dia-illumination
Average lamp life 100 hours
Light power supply 6V30W integrated (100 to 240V)
Input ratings 100-240VAC ±10%, 50/60Hz, 0.8A
Power consumption 38W
(nominal)
Power cord • When used in 100-120 V regions outside Japan
UL listed detachable power cord set, 3 conductor grounding
(3 conductor grounding Type SVT, No.18 AWG, 3 m long maximum, rated at 125 VAC
minimum)
• When used in 220-240 V regions
Detachable power cord set approved in accordance with EU/EN standard, 3 conductor
grounding
(3 conductor grounding Type H05VV-F, 3 m long maximum, rated at 250 VAC minimum)
• When used inside Japan
PSE approved detachable power cord set, 3 conductor grounding Chapter 6
2
(3 conductor grounding Type VCTF 3 x 0.75 mm , 3 m long maximum, rated at 125 VAC
minimum)

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 Chapter 6 Specifications and Safety Standards

■ Nikon Microscope ECLIPSE Ci-L

Model ECLIPSE Ci-L

Optical system Infinity-corrected CF optical system
Objective CFI60
Eyepiece: Field number 22 (with ergonomic tube/binocular tube),
25 (with T/F trinocular tube)
Nosepiece: Sextuple
Focus up/down motion Drive system: Manual coarse/fine motion
(calibration markings for fine motion: 1 µm/marking)
Stroke: 2 mm upward, 28 mm downward
With refocusing mechanism
Light source for White LED
dia-illumination
Light power supply 5V15W integrated (100 to 240V)
Input ratings 100-240VAC ±10%, 50/60Hz, 0.37A
Power consumption 6W
(nominal)
Power cord • When used in 100-120 V regions outside Japan
UL listed detachable power cord set, 3 conductor grounding
(3 conductor grounding Type SVT, No.18 AWG, 3 m long maximum, rated at 125 VAC
minimum)
• When used in 220-240 V regions
Detachable power cord set approved in accordance with EU/EN standard, 3 conductor
grounding
(3 conductor grounding Type H05VV-F, 3 m long maximum, rated at 250 VAC minimum)
• When used inside Japan
PSE approved detachable power cord set, 3 conductor grounding
2
(3 conductor grounding Type VCTF 3 x 0.75 mm , 3 m long maximum, rated at 125 VAC
minimum)

■ CI-FL Epi-fluorescence Attachment for Nikon Microscope

Model CI-FL epi-fluorescence attachment

Optical system Infinity-corrected CF optical system
Variable intermediate magnification: 1x
Specifications and Safety Standards

Epi-fl filter turret Manual quadruple turret

Field diaphragm Manual adjustment
ND filter Manual, 3 filters (slider type; ND4, ND8, ND16)
Shutter Manual (using a lever at the bottom of the turret)
Supported illuminator HG Precentered Fiber Illuminator

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 Chapter 6 Specifications and Safety Standards

3 Physical Properties

■ Nikon Microscope ECLIPSE Ci-S

Model ECLIPSE Ci-S

Operating conditions Temperature: 0°C to +40°C
Humidity: 60% RH max. (no condensation)
Altitude: 2000 m max.
Pollution degree: Degree 2
Installation: Category II
Electrical shock protection class: Class I
Indoor use only
Transport/storage Temperature: -20°C to +60°C
conditions Humidity: 90% RH max. (no condensation)
External dimensions and External dimensions: 223 (W) x 331.5 (H) x 331 (D) mm
weight (Main body only) (excluding projections)
Weight: Approx. 10 kg

■ Nikon Microscope ECLIPSE Ci-L

Model ECLIPSE Ci-L

Operating conditions Temperature: 0°C to +40°C
Humidity: 60% RH max. (no condensation)
Altitude: 2000 m max.
Pollution degree: Degree 2
Installation: Category II
Electrical shock protection class: Class I
Indoor use only
Transport/storage Temperature: -20°C to +60°C
conditions Humidity: 90% RH max. (no condensation)
External dimensions and External dimensions: 223 (W) x 331.5 (H) x 331 (D) mm
weight (Main body only) (excluding projections)
Weight: Approx. 10 kg

Specifications and Safety Standards

■ CI-FL Epi-fluorescence Attachment for Nikon Microscope

Model CI-FL epi-fluorescence attachment

Operating conditions Temperature: 0°C to +40°C
Humidity: 60% RH max. (no condensation)
Altitude: 2000 m max.
Pollution degree: Degree 2
Installation: Category II
Electrical shock protection class: Class I
Indoor use only
Transport/storage Temperature: -20°C to +60°C
conditions Humidity: 90% RH max. (no condensation)
Mass Approx. 2kg

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