Upright Microscope for Routine and Entry-level Research
Page 2
ZEI
SS Copyright / Trademarks Axioscope 5/7/Vario
Knowledge of this manual is essential for the operation of the instrument. Please familiarize yourself with
the contents of this manual and pay special attention to instructions concerning safe operation of the
instrument.
The specifications are subject to change; the manual is not covered by an update service.
commercial exploitation and communication of its contents are not permitted. Persons in
contravention of this copyright are liable to pay compensation for damages.
All rights reserved, including rights created by patent grant or registration of a utility model.
ll names of companies and products mentioned in this manual may be trademarks or registered
A
trademarks. Third party products are cited for information purposes only and this does not represent
approval or recommendation of these products.
Carl Zeiss Microscopy GmbH assumes no liability for the performance or use of such products.
authorized dealer:
Pulch + Lorenz microscopy
Am Untergrün 23, D-79232 March
tel:07665 9272-0
fax:07665 9272-20
mail: kontakt@pulchlorenz.de
web: pulchlorenz.de
Published by Carl Zeiss Microscopy GmbH
Carl-Zeiss-Promenade 10
07745 Jena, Germany
microscopy@zeiss.com
www.zeiss.com/microscopy
Carl Zeiss Suzhou Co., Ltd.
Modern Industrial Square 3-B, No.333
XingPu Road SIP
215126 Suzhou, China
O
rder No.: 430035-7344-001
Issue date: Version 3 – 15 March 2018
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A
xioscope 5/7/Vario Contents / List of Illustrations ZEISS
Fig. 4-19 Components for reflected light polarization ..................................................................... 121
Fig. 4-20 Components for reflected light fluorescence .................................................................... 123
Fig. 5-1 Replacing the fuses in the stand ...................................................................................... 126
Fig. 5-2 Replacing fuses in the power supply unit .......................................................................... 127
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INTRODUCTION
Axioscope 5/7/Vario
Notes on instrument safetyZEISS
1 INTRODUCTION
1.1 Notes on instrument safety
The Axioscope 5/7/Vario microscopes were engineered, manufactured and tested in accordance with the
DIN EN 61010-1 (IEC 61010-1) and IEC 61010-2-101 safety standards for electrical measuring, control
and laboratory equipment.
The microscopes fulfill the requirements as stated in Directive 98/79/EC on in vitro diagnostic medical
devices (IVDD) and bear the marking.
The present manual contains information and safety warnings with which the operator must comply.
The following warning, instructional and informational symbols are used in the manual:
CAUTION
Warns the operator of a possible danger.
CAUTION
Warns the operator of hot surfaces.
CAUTION
Warns the operator of the emission of UV radiation.
CAUTION
Warns the operator of the emission of LED radiation.
LED class 3B, max. 30 mW, 365 - 625 nm,
Avoid direct exposure to the beam. Avoid contact with the skin.
CAUTION
Warns the operator of crushing hazard.
CAUTION
Warns the operator to disconnect the instrument from the power supply before opening or
doing any work on the instrument.
ATTENTION
This symbol indicates the risk of damage to the instrument or the system.
NOTE
This symbol indicates information or instructions which must be followed especially carefully.
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INTRODUCTION
out of order, the
Microscopy Service before
100 covers a voltage range from
Hz. It
ake sure that the fuses are suitable for the applied nominal current. Never use any
hazardous to your
ZEISS Notes on instrument safety Axioscope 5/7/Vario
The Axioscope 5/7/Variomicroscopes and their original accessories may be used only for the microscopy
procedures described in this operating manual.
The following guidelines are especially important:
CAUTION
The microscope may only be plugged into an electrical outlet equipped with a safety contact.
The safety feature must not be disabled by using an extension cord which does not have a
protective ground conductor.
CAUTION
Whenever it becomes apparent that any of the safety mechanisms are
microscope must be switched off and protected from any inadvertent use.
Please contact the ZEISS Service Department or the Carl Zeiss
switching the microscope on again.
CAUTION
The stands are equipped with integrated power supply units. These units adapt to a voltage
range of 100 to 240 V and a frequency range of 50 to 60 Hz. No further voltage adjustment
is necessary on the microscope itself.
The 12 V DC 100 W external power supply unit for HAL
100 to 127 V AC and 200 to 240 V AC ±10%, and a frequency range of 50 to 60
adapts automatically to the applied voltage.
The 24 V DC 30 W external power supply unit for Axioscope 5 Vario covers a voltage range
from 100 V to 240 V and a frequency range of 50 to 60 Hz.
The power supply units for the HBO 100 adapt to a voltage range from 100 to 240 V AC and
a frequency range of 50 to 60 Hz. They adapt automatically to the applied voltage. No further
voltage adjustment is necessary.
Before you switch on the instrument, please verify that the voltage you are about to use is
suitable.
CAUTION
Always disconnect the microscope from the power supply before you open it or change a
fuse!
M
makeshift fuses and do not short-circuit the fuse holders.
CAUTION
The microscopes do not have any special safety devices to protect users from acid or from
potentially infectious, toxic, radioactive or other samples that may be
health. Compliance with all statutory requirements, especially national accident prevention
regulations, is required when handling such samples.
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INTRODUCTION
using the microscope. The
can easily be damaged or even
to the beam. Never operate the microscope without the necessary safety equipment (e.g.
ilters or the fluorescence protection screen). Gas discharge lamps have a
disconnect the
white light source coupled by a fluid light conductor is used, ultraviolet radiation may leak
he light and avoid
with or without optical
even if you just want to look at a specimen. Irreparable eye damage may
Axioscope 5/7/Vario
Notes on instrument safetyZEISS
CAUTION
Only authorized personnel are permitted to operate the instruments. The personnel must have
been instructed about and be aware of the risks involved in
Axioscope is a precision instrument whose functionality
destroyed when handled improperly.
CAUTION
Operating the instrument in an area with a potentially explosive atmosphere is prohibited.
It may be operated only on stable, non-flammable surfaces.
Specimens must be disposed of appropriately in accordance with applicable statutory
regulations and internal work instructions.
CAUTION
Gas discharge lamps, such as the HBO 100, emit ultraviolet radiation which may cause injuries
to your eyes and skin. Never look directly into the beam and avoid exposing your skin directly
special damping f
high internal pressure when hot. Never change the lamps while they are still hot and always
use protective gloves and masks when changing them.
CAUTION
Avoid any contact with the hot lamp case. Before you change a lamp,
instrument from the power supply and wait 15 minutes for the lamp to cool down.
CAUTION
When the lamp module (e.g. Colibri 3) is equipped with appropriate LED modules, or when a
which can cause injury to your eyes and skin. Never look directly into t
exposing your skin directly to the light. Always use the appropriate safety equipment when
operating the microscope.
CAUTION
Never look into the light beam of the illumination equipment instruments - not
occur!
CAUTION
Do not hold any flammable or easily combustible materials into the light beam.
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INTRODUCTION
immediately for at least 5 minutes. Seek medical
appropriately: Do not allow to contaminate
assemblies or
Moreover, the warranty and any claims
are always kept open,
ZEISS Notes on instrument safety Axioscope 5/7/Vario
CAUTION
The immersion fluid Immersol 518 N is irritating to the skin. Avoid any contact with skin,
eyes and clothes. Read the safety data sheets on Immersol 518 N, Immersol 518 F and
Immersol W carefully. If skin contact occurs, wash off with lots of water and soap.
If eye contact occurs, flush with water
assistance if irritation continues.
CAUTION
Dispose of immersion fluid Immersol 518 N
surface water or get into drains.
CAUTION
For stands equipped with motorized focusing drives, there is a risk of fingers being crushed in
the working area when the microscope stage is lowered.
Do not reach into the area between the microscope stage/stage carrier/condenser carrier and
the base of the stand while the stage is being lowered.
ATTENTION
The manufacturer assumes no liability for any use of the microscope, its
individual parts if these are used in in a manner or for any purpose other than those described
in this operating manual. This also applies to any maintenance or repair work not performed
by authorized maintenance and repair personnel.
thereunder will be voided.
ATTENTION
Microscope lamps radiate a lot of heat which could damage the heat-sensitive fluorescence
filters. Do not remove the heat protection filter when using a fluorescence filter; otherwise the
fluorescence filter may not function properly.
ATTENTION
Dirt and dust may affect the microscope’s performance. Protect it by using a dust cover when
it is not in use. Always make sure that the instrument is switched off before covering it up.
ATTENTION
Closing or covering the ventilation slits may result in heat accumulation which could damage
the instrument and even start a fire. Ensure that the ventilation slits
uncovered and clear. Do not insert or drop anything into them or let anything fall into them.
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INTRODUCTION
in accordance with applicable statutory
Axioscope 5/7/Vario
Warning labels on the microscopesZEISS
ATTENTION
Always position equipment in a manner that allows easy disconnection from the power
supply.
ATTENTION
Do not use a power supply cord with an inadequate rating.
ATTENTION
When using the HAL or HBO illuminators, a base plate for the Axioscope (000000-2202-526)
is mandatory.
ATTENTION
Do not dispose of defective microscopes in the household waste. Comply with the applicable
statutory regulations for their disposal.
Specimens must also be disposed of appropriately
regulations and internal work instructions.
1.2 Warning labels on the microscopes
Fig. 1-1 Warning labels on microscopes with LED light source for transmitted light
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INTRODUCTION
ZEISS Warning labels on the microscopes Axioscope 5/7/Vario
Fig. 1-2 Warning labels on microscopes with HAL 100 for transmitted light and HBO 100 for reflected
light
Fig. 1-3 Warning label on microscopes with LED light source for reflected light
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INTRODUCTION
Axioscope 5/7/Vario
Warning labels on the microscopesZEISS
Fig. 1-4 Warning label on microscopes with Colibri 3 light source for reflected light
Fig. 1-5 Warning label on Axioscope 7 stand using the motorized 80x60 mechanical stage 80x60
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INTRODUCTION
ZEISS Notes on the warranty Axioscope 5/7/Vario
1.3 Notes on the warranty
The manufacturer guarantees that the instrument is free of any material and workmanship defects upon
delivery. If you become aware of any deficiencies, please contact us immediately and take all necessary
precautions in order to avoid further damage. Upon notice of deficiencies it is up to the manufacturer’s
discretion to correct the deficiencies or to deliver a defect-free instrument. Defects due to ordinary wear
and tear (especially on wearing parts) and to improper handling are not covered by our warranty.
The manufacturer is not liable for damage to the instrument due to incorrect operation, negligence or
any other manipulation of the instrument, in particular due to the removal or replacement of instrument
parts or due to the use of accessories from other manufacturers. This will immediately void the warranty.
No maintenance or repair work, except for the instances mentioned in the manual, may be performed on
the microscopes. Only ZEISS service personnel or personnel authorized especially by ZEISS may perform
repair work on the microscopes. In the event of malfunction of your instrument, please contact the ZEISS
Microscopy Service Department (see page 131) or the ZEISS agency assigned to your country.
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DESCRIPTION OF THE INSTRUMENT
Transmitted light
Reflected light
−
−
−
−
−
−
−
−
−
−
−
−
Axioscope 5/7/Vario
Intended useZEISS
2 DESCRIPTION OF THE INSTRUMENT
2.1 Intended use
The Axioscope 5/7/Vario microscopes are universal microscopes designed especially for applications in
biological and medical fields and for materials analysis.
Depending on the configuration of the microscope stand, they may be used with transmitted light only or
with a combination of transmitted light and reflected light.
Typical areas of use for the Axioscope 5/7/Vario in biomedical applications include:
− medical analysis in laboratories, hospitals, physicians’ practices,
− science and research (colleges, universities) in medical and biological fields,
The binocular photo tubes can be connected via a special adapter with a microscopy camera, a reflex
camera or a digital/video camera for documentation purposes. An Infinity Space Camera Port (ISCP)
camera can be used when working with transmitted light.
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DESCRIPTION OF THE INSTRUMENT
ZEISS Technical data Axioscope 5/7/Vario
2.2 Technical data
Dimensions (length x width x height)
Microscope stand for Axioscope 5/7 ......................................... approx. 293.5 mm x 240 mm x 367.5 mm
Microscope stand for Axioscope Vario ......................................... approx. 429 mm x 458.5 mm x 700 mm
Weight
Microscope stand for Axioscope 5/7 ........................................................................................................
(depending on stand configurations and accessories) .................................................. approx. 14 to 20 kg
Microscope stand for Axioscope Vario ................................................................................. approx. 32 kg
Ambient conditions
Shipping (in packaging):
Permissible ambient temperature .................................................................................... -40 to +70 °C
Permissible humidity (no condensation) ................................................................. max. 75% at 35 °C
Storage:
Permissible ambient temperature ................................................................................... +10 to +40 °C
Permissible humidity (no condensation) ................................................................. max. 75% at 35 °C
Operation:
Permissible ambient temperature ................................................................................... +10 to +40 °C
Permissible relative humidity (no condensation) ...................................................... max. 75% at 35 °C
Highest permitted altitude of use .................................................................................... max. 2000 m
Air pressure ........................................................................................................ 800 hPa to 1060 hPa
Degree of pollution ........................................................................................................................... 2
Operational specifications
Operational area ................................................................................................................ Closed rooms
Protective class ....................................................................................................................................... I
Protection type ................................................................................................................................ IP 20
Electrical safety............................................................ in accordance with DIN EN 61010-1 (IEC 61010-1)
............................................................................................ in conjunction with CSA and UL regulations
Overvoltage category............................................................................................................................. II
RFI suppression....................................................................................... conforming to EN 55011 Class B
Noise immunity ..................................................................................... conforming to DIN EN 61326/A1
Mains voltage for the Axioscope 5/7
with internal power supply unit ....................................................................................... 100 to 240 V
Mains voltage for the Axioscope Vario
with external power supply unit ...................................................................................... 100 to 240 V
Mains frequency .................................................................................................................... 50 to 60 Hz
Power consumption of the Axioscope 5
with internal power supply unit ................................................................................................. 60 VA
Power consumption of the Axioscope 7
with internal power supply unit ............................................................................................... 100 VA
Power consumption of the Axioscope Vario
with external power supply unit ................................................................................................ 30 VA
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DESCRIPTION OF THE INSTRUMENT
Axioscope 5/7/Vario
Technical dataZEISS
HBO 100 W power supply unit
Range of application ..................................................................................................................... indoor
Protection class .......................................................................................................................................I
Protection type ................................................................................................................................ IP 20
Mains voltage ........................................................................................................ 100 VAC ... 240 VAC
............................................................................................................... No voltage adjustment required
Mains frequency ................................................................................................................... 50 to 60 Hz
Power consumption with HBO 103 in operation ........................................................................... 155 VA
Fuses in accordance with IEC 127
Axioscope 5/7 microscope stand for LED illumination in transmitted light: ............. 2 T 3.15 A/H, 5x20 mm
Axioscope 5/7 microscope stand for HAL 50 illumination in transmitted light: ....... 2 T 3.15 A/H, 5x20 mm
HBO 100 W power supply unit ................................................................................. T 2.0 A/H, 5x20 mm
12 V DC 100 W external power supply unit ............................................................ 2 T 5.0 A/H, 5x20 mm
Light sources
LED illumination transmitted light/reflected light
Power consumption ........................................................................................................... max. 10 W
Adjustment of light source .............................................................. continuous approx. 10 to 800 mA
Halogen lamp ......................................................................................................................... 12 V/50 W
Adjustment of light source .................................................... infinitely variable from approx. 3 to 12 V
Halogen lamp ....................................................................................................................... 12 V/100 W
Adjustment of light source .................................................... infinitely variable from approx. 3 to 12 V
Mercury vapor short arc lamp............................................................................................. HBO 103 W/2
Power consumption for HBO 103 W/2 ...................................................................................... 100 W
Colibri 3 illumination system
Power consumption ................................................................................................................... 15 W
modulator disk ...................................................................................... for bright-field, dark-field and
........................................................................................................ phase contrast 1, 2, 3 or PlasDIC
Manual objective change ........................................................................ via nosepiece, 6-positions, M27
Manual reflector module change .................................................... via reflector slider, 2-positions coded,
............................................................................. reflector turret 4-positions coded or 6-positions coded
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DESCRIPTION OF THE INSTRUMENT
ZEISS Interface diagram Axioscope 5/7/Vario
2.3 Interface diagram
The following figure shows a diagram of the microscope’s hardware interfaces. The Axioscope 5 stand
Bio-TL/RL with LED/HAL illumination serves as the example.
Controls and functional elements on the microscopeZEISS
Key to 284HFig. 2-1:
1Reflector turret
2Upper part of the stand
3Camera interface 60N on the photo tube
4Tube/photo tube
5Filter slider for reflected light
6Adjustment tool
7Lower part of the stand
8Filter slider for transmitted light
9Filter wheel for transmitted light
10Condenser carrier
11Condenser
12Stage carrier
13Stage
2.4 Controls and functional elements on the microscope
2.4.1 Stand models
Eight stand models are available in the delivery program:
8. Axioscope 5 Vario material stand (430035-9150-000)
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DESCRIPTION OF THE INSTRUMENT
ZEISS Controls and functional elements on the microscope Axioscope 5/7/Vario
2.4.2 Axioscope 5 stands, Bio-TL
Transmitted light stands for bioscience equipped with the following main components:
1. Version 1 (430035-9032-000): HAL 50 halogen illumination
Version 2 (430035-9201-000): LED 10 W-TL illumination or optionally
HAL 100 halogen illumination using an external power supply
7. Coded changeable reflector turrets and reflector slider
Key to Fig. 2-2:
1Eyepieces
2Binocular tube part
3Binocular tube/photo tube (with mounted Axiocam 208, optionally)
4Transmitted light stand
5Transmitted light illuminator
6Intensity/LM knob for light intensity and Light Manager function (LM)
7Indicator light for transmitted light
8Permanent/ECO mode switch
9Filter slider transmitted light 10Focusing drive – fine adjustment (left & right)
11Focusing drive – coarse adjustment (left & right)
12Coaxial knurled knob for adjusting the mechanical stage in direction Y
13Coaxial knurled knob for adjusting the mechanical stage in direction X
14Snap buttons
15Nosepiece 166-position filter wheel (operable from left & right)
17Field diaphragm
18Condenser with aperture diaphragm and centering screws (with optional modulator disk)
19Knurled knob for adjusting the condenser vertically (left & right)
20Stage carrier
21Condenser carrier
22Release lever for height stop on focusing drive
23Main switch I/O
24Mechanical stage with dual specimen holder
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DESCRIPTION OF THE INSTRUMENT
Axioscope 5/7/Vario
Controls and functional elements on the microscopeZEISS
Fig. 2-2 Controls and functional elements of the Axioscope 5 stands, Bio-TL
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DESCRIPTION OF THE INSTRUMENT
ZEISS Controls and functional elements on the microscope Axioscope 5/7/Vario
2.4.3 Axioscope 5 stand, Bio-TL/RL
Transmitted and reflected light fluorescence stand for bioscience equipped with the following main
components:
1. LED 10 W-TL illumination or optionally with HAL 100 halogen
illumination using an external power supply for transmitted light
applications
6. Coded changeable reflector turret or reflector slider
7. Stand column 560 mm with crank for vertical adjustment
8. Stage carrier and base plate
Key to Fig. 2-7:
1Eyepieces
2Binocular tube part
3Binocular tube/photo tube (with mounted Axiocam 208, optionally)
4Release lever for vertical adjustment
5Stand column Axioscope Vario 560 mm
6Hand wheel for vertical adjustment
7Slot for polarizer slider A 60x30 mm
8Field diaphragm (as slider)
9Aperture diaphragm (as slider)
10Filter slider for reflected light
11Reflected light illuminator
12Intensity/LM knob for light intensity and Light Manager function (LM)
13Focusing drive – coarse adjustment
14Focusing drive – fine adjustment
15Snap button
16Indicator light for reflected light
17Permanent/ECO mode switch
18Base plate
19Stage carrier, H=140 mm
20Nosepiece
21Knurled wheel for switching the reflector turret position
22Reflector turret (for replaceable reflector inserts)
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DESCRIPTION OF THE INSTRUMENT
Axioscope 5/7/Vario
Controls and functional elements on the microscopeZEISS
Fig. 2-7 Controls and functional elements of the Axioscope 5 Vario material stand
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DESCRIPTION OF THE INSTRUMENT
Axioscope 7
mot. Axioscope 5 Axioscope
Vario
ZEISS Controls and functional elements on the microscope Axioscope 5/7/Vario
2.4.8 Functions of stands keys and display elements
For the location of the keys and display elements on your stand, refer to sections 2.4.2 - 2.4.7.
NOTE
Each time the microscope is switched on, the first setting of parfocal value for any nosepiece
position will clear the parfocal settings for ALL nosepiece positions.
Availability
Key
Main switch I/O
(see Fig. 2-6/29)
Permanent/
ECO switch
(see Fig. 2-6/16)
Indicator light
(see Fig. 2-6/12
and 15)
RL/TL button
(see Fig. 2-6/13
and 14)
Intensity/
LM knob
(see Fig. 2-6/10)
* Short press: hold less than 1 second, then release.
** Long press: hold at least 1.5 seconds.
*** BLINK: the indicator light alternately goes on/off at 500 ms intervals.
Yes Yes Yes I = on; O = off Switches the microscope on/off
Yes Yes Yes Switch:
Yes Yes Yes Blinking ***: Indicates microscope function (normal/setting)
Yes Option No Short press *:
Yes Yes Yes
5
Action Functionality/Description
Switches between Permanent (continuous) mode and
ECO mode of the microscope illumination:
- Permanent mode active: illumination is
continuously switched on.
- ECO mode active: illumination switches off after
15 minutes without action.
Switches RL/TL illumination alternately on/off.
The respective indicator light is continuously
illuminated. Second press to the RL/TL button to turn
off/on the illumination (the indicator light not affected).
Turn: Controls the light intensity of the active light source
Repeated short pressing switches a single LED or all
Short press *:
Long press **:
Long press for
20 s:
LEDs of the Colibri 3 together on or off (if Colibri 3 is
connected).
Saves the set light intensity. During this action, the
indicator light blinks twice in GREEN and the image
background appears BLACK for 300 ms (this does not
apply to halogen illumination).
Activates the factory default settings (enables Light
Manager, sets light intensity to the initial value, clears
all saved parfocal positions).
When the knob is pressed, the indicator light starts
blinking in RED after 3 s until 20 s is reached. After
20 s, the indicator light blinks in GREEN. Then release
the knob. The indicator light turns to GREEN
permanently if the system reset is done.
After factory default reset, need re-power on the
system.
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DESCRIPTION OF THE INSTRUMENT
Axioscope 7
mot. Axioscope 5 Axioscope
Vario
Axioscope 5/7/Vario
Controls and functional elements on the microscopeZEISS
Availability
Key
Left Snap
button (see
Fig. 2-5/22)
(if Axiocam
attached)
Right Snap
button (see
Fig. 2-6/20)
(if Axiocam
attached)
Snap button +
Intensity/LM
knob (see Fig. 2-6/20+10)
Stage control
button on the
left
(see Fig. 2-6/28)
Right snap
button + Stage
control button
on left (see
Fig. 2-6/20+28)
* Short press: hold less than 1 second, then release.
** Long press: hold at least 1.5 seconds.
*** BLINK: the indicator light alternately turns on/off at 500 ms intervals
No Yes No
Yes Yes Yes
Yes Yes Yes Long press **:
Yes No No
Yes No No
5
Action Functionality/Description
Short press *: Snaps an image
Long press **:
Short press:
Long press **:
Short press *:
Long press **:
Press
simultaneously
Press
simultaneously
again
Starts video recording; another short press is required
to stop recording.
Snaps an image; when the snap is completed, the
monitor display turns BLACK for 50 ms.
Starts video recording; another short press is required
to stop recording.
Enables/disables the Light Manager:
- Disabling: The indicator light blinks
GREEN / ORANGE / GREEN in sequence.
- Enabling: The indicator light blinks
GREEN / GREEN / GREEN in sequence.
Switches between XY stage control and Z axis control
via focus drives
If Z axis control is active:
- the indicator light lights in GREEN
- the left or right fine focusing drive knobs are
controlled for slow Z-movement (focusing)
- the left and right coarse focusing drive knobs
are controlled for fast Z-movement (focusing)
If XY stage control is active:
- the indicator light blinksORANGE
- the left focus drive knobs (fine or coarse)
control Y movement (slow or fast) of the
stage
- the right focus drive knobs (fine or coarse)
controls X movement of (slow or fast)
the stage
Records the parfocal position.
1. the LED shuts off for 300 ms (darkness indicates
action to user)
For halogen illumination: Do NOT shut off
the light.
2. the indicator light blinks twice in GREEN
Load/unload
Load/unload
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DESCRIPTION OF THE INSTRUMENT
ZEISS Controls and functional elements on microscope components Axioscope 5/7/Vario
2.5 Controls and functional elements
on microscope components
2.5.1 Binocular photo tube 30°/23
(100:0/0:100)
The photo port (
710HFig. 2-8/2) of the binocular tube
can be used to mount a reflex camera, a
microscope camera or a video camera with the aid
of appropriate adapters.
Using the shift knob (or the push-pull rod on other
photo tubes), the light can be directed to the
eyepieces or to the mounted camera.
Fig. 2-8 Binocular photo tube
− Shift knob (
(eye symbol):
100% light to the eyepieces
− Shift knob (
(camera symbol):
711HFig. 2-8/3) in forward position
712HFig. 2-8/3) in back position
100% light to the camera
− Push-pull rod (
713HFig. 2-8/1) pushed in:
Eyepiece shutter is closed.
− Push-pull rod (
714HFig. 2-8/1) pulled out:
Eyepiece shutter is open.
− Whenever you want to take photos with a long exposure time, we recommend that you block any
ambient light from entering the eyepieces by using the tube shutter or the eyepiece cover (included
in the dust protection kit). If neither is available, remove the eyepiece and place the included dust
protection caps over the eyepiece barrels.
− Vary eyepiece distance by moving the eyepiece barrels apart or together.
NOTE
For polarization microscopy, we recommend using the Pol photo tube with upright image and
one eyepiece reticle (graticule).
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DESCRIPTION OF THE INSTRUMENT
Axioscope 5/7/Vario
Controls and functional elements on microscope componentsZEISS
2.5.2 Binocular ergo tube/ergo photo
tube 20°/23 and ergo photo tube
15°/23, each with continuous
vertical adjustment
These ergonomic tubes are designed for a field of
view of 23 mm. The viewing angle is 20° or 15°.
The ergo photo tubes are equipped with one
camera port each (Fig. 2-9/1 and Fig. 2-10/2).
The camera ports can be used to mount a reflex
camera, a microscope camera or a video camera
with the aid of appropriate adapters.
The camera port can carry a maximum weight of
2.5 kg (camera plus cable). For a fee, the ZEISS
service personnel will equip your instrument with a
higher maximum load, if needed.
Fig. 2-9 Binocular ergo photo tube 20°/23
with vertical adjustment
The ergo tubes permit a height adjustment range
of 44 mm.
A larger range can be obtained by swiveling the
binocular section from the lower to the upper
observation position (depending on the interpupillary distance).
− Continuous vertical adjustment by means of
the rotary knobs (Fig. 2-9/3 and Fig. 2-10/1)
− The adjustment range of the ergo photo
tube 20°/23 can be read off the vertical
scale (Fig. 2-9/4).
−The binocular part of the ergo photo tube
15°/23 is also continuously horizontally
retractable up to 50 mm.
Fig. 2-10 Binocular ergo photo tube 15°/23,
telescopic with vertical adjustment
The ergo photo tube 20°/23 has two switch positions (scale: 100:0/0:100):
− Push-pull rod (Fig. 2-9/2) pushed in: 100% to the eyepieces
− Push-pull rod (Fig. 2-9/2) pulled out: 100% to camera port
The ergo photo tube 15°/23 (Fig. 2-10) is available only with an upright image and fixed scale (50:50).
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DESCRIPTION OF THE INSTRUMENT
ZEISS Controls and functional elements on microscope components Axioscope 5/7/Vario
2.5.3 Nosepiece with objectives
Fig. 2-11 Nosepiece with DIN slot for
compensators
− Equipped with DIN slot (
compensators or fluorescence protection
shield
− M27 mounting thread for six objectives
− Equipped with three, six or no DIC positions
(
704HFig. 2-11/1) depending on the configuration
− Turning the knurled wheel of the nosepiece
(
705HFig. 2-11/2) enables you to change the
objectives quickly
2.5.4 Mechanical stage 75x50
− Mechanical stage (
holder (
718HFig. 2-12/2) for holding and
717HFig. 2-12/1) with specimen
positioning the samples
703HFig. 2-11/3) for
Fig. 2-12 Mechanical stage with specimen
holder
− Two clamping screws (
717HFig. 2-12/3) for
attaching and removing the specimen holder
− Coaxial knurled knobs for X (
Y adjustment (
720HFig. 2-12/5)
719HFig. 2-12/4) and
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DESCRIPTION OF THE INSTRUMENT
Axioscope 5/7/Vario
Controls and functional elements on microscope componentsZEISS
2.5.5 Low-power system for 2.5x/4x
objectives mounted on the
condenser carrier
The low-power system enables full illumination of
the field of view when using a low-magnification
objective (2.5x-4x) in combination with the
condenser 0.9/1.25 H.
It can be centered and remains in the beam path
throughout the use of the corresponding objective.
• Swing the low-power system (
721HFig. 2-13/2) in or
out of the beam path using the handle
722HFig. 2-13/1). Make sure the module locks in
(
securely.
• Do not use the low-power system with LED
illumination for transmitted light.
ATTENTION
If the condenser carrier with mounted low-power system is moved too far down, the lowpower system may collide with the field diaphragm and damage it!
2.5.6 0.9/1.25 BF condenser and 0.9 Pol
condenser
The 0.9/1.25 BF condenser (
aperture diaphragm (
701HFig. 2-14/2) can be optionally
equipped with a modulator disk (
700HFig. 2-14/1) with
702HFig. 2-14/3) for:
Fig. 2-13 Low-power system mounted on the
condenser carrier
− bright-field BF
− dark-field DF
− phase contrast PhC 1, PhC 2, PhC 3
− PlasDIC (optional: To use PlasDIC, the ring
diaphragm must be replaced with a 2 mm or
Fig. 2-14 0.9/1.25 BF condenser with
modulator disk
4.5 mm slit diaphragm for PlasDIC)
ATTENTION
Do not use acetone to clean the lens of the polarizing condensers.
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DESCRIPTION OF THE INSTRUMENT
installed) on the level of the aperture diaphragm in order to attenuate the fluorescence
ZEISS Controls and functional elements on microscope components Axioscope 5/7/Vario
2.5.7 14x40 mm stop sliders for aperture
and field diaphragms
To adjust the reflected light beam path using the
KÖHLER method, one 14x40 mm stop slider
723HFig. 2-15/4) mounted in the upper part of the
(
stand is required to function as a field diaphragm
(F) and another to function as the aperture
diaphragm (A).
• To open the diaphragm, turn the knurled wheel
(
724HFig. 2-15/1) up.
• To close the diaphragm, turn the knurled wheel
(
Fig. 2-15 14x40 mm stop slider
724HFig. 2-15/1) down.
• To center the diaphragm to the beam path,
adjust the centering screws (
725HFig. 2-15/2 and 3)
with an Allen wrench (AF 3).
NOTE
When applying fluorescent light, you may want to mount an FL attenuator (if not pre-
radiation.
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DESCRIPTION OF THE INSTRUMENT
Axioscope 5/7/Vario
Controls and functional elements on microscope componentsZEISS
2.5.8 Reflector turrets with 4 or 6 coded
positions
These reflector turrets are equipped with four or
six coded push-and-click (P&C) reflector positions.
Turn the knurled wheel to adjust the reflector
position. The marking indicates which reflector
position is currently in the beam path.
For easy identification we advise that you write
down the appropriate data for each position of the
mounted reflector modules on the adhesive labels
provided with the instrument. Then attach the
labels at the appropriate points.
1Display showing the number of the active
reflector position
2Spaces for labels with position data
3Knurled wheel with position marks
2.5.9 Reflector slider with 2 coded
positions
The reflector slider with two coded positions is
equipped with two individually loadable reflector
positions for P&C modules which can be slid into
the beam path.
Fig. 2-16 Reflector turret with 6 coded
positions
1 Slider
Fig. 2-17 Reflector slider 2 coded positions
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ZEISS Mounting standard components Axioscope 5/7/Vario
3 START-UP
The Axioscope microscopes can be installed, modified and started up by the customer himself.
On request, the ZEISS Service installs or modifies the microscope for an extra charge.
NOTE
Before installing and starting-up the microscope, read the Notes on instrument safety (see
section 1.1) carefully and thoroughly.
The assembly activities described on the following section are illustrated for the most part by way of
examples for one microscope stand type. However, they apply analogously to other microscope stand
models. Special features are described separately.
The same activities apply when assembling a microscope stand with an upper stand part and the stand
column Vario – special activities for the stand column Vario are described separately.
Fig. 3-1 Setting up the microscope
3.1 Mounting standard components
3.1.1 Unpacking and setting up
microscope stand
• Remove all components from the packaging
and check that all components described on the
delivery note are present.
• Set up the microscope stand (
vibration-free, level, hard and non-combustible
surface.
• Keep the original packaging for storage or for
returning the instrument to the manufacturer,
or dispose of it properly.
286HFig. 3-1/1) on a
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Mounting standard componentsZEISS
3.1.2 Mounting the Axioscope 5 Vario material upper stand part on the stand column
ATTENTION
Equipment or parts having a mass of 18 kg or more shall be provided with a means for careful
lifting and carrying.
If you are going to use the Axioscope 5 Vario material stand consisting of the Vario upper stand part
(288HFig. 3-2/2) combined with gear box (288HFig. 3-2/3) and the Vario stand column (288HFig. 3-2/1), proceed as
follows:
• Unpack the Vario upper stand part with gear box and the Vario stand column.
• Position the Vario upper stand part (
column and tighten it with the 6 Allen screws (
• In order to adjust the microscope vertically, loosen the release lever (
287HFig. 3-2/2) on the mounting plate (288HFig. 3-2/5) of the Vario stand
287HFig. 3-2/4).
292HFig. 3-2/6) on the stand column
and adjust the height (according to the size of the specimen) with the hand wheel (
• Then tighten the release lever again.
292HFig. 3-2/7).
Fig. 3-2 Mounting the Vario upper stand part on the stand column
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ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.3 Mounting the binocular
tube/photo tube
Fig. 3-3 Attaching the binocular tube
• Loosen the screws (
295HFig. 3-3/3) with the Allen
wrench (AF 3).
• Remove the dust cover caps (
296HFig. 3-3/2 and 5)
from the bottom of the tube and the dovetail
ring socket on the stand.
• Hold the binocular tube (
297HFig. 3-3/1) or the
binocular photo tube at an angle and insert it
into the stand socket with the dovetail ring
(
298HFig. 3-3/4).
• Then push it into a horizontal position. Turn the
binocular tube into the desired viewing position
and tighten the Allen screws with the Allen
wrench.
3.1.4 Inserting eyepieces or auxiliary
microscope or pinhole diaphragm
Fig. 3-4 Inserting eyepieces
• Remove both dust cover caps (
299HFig. 3-4/1 and 5)
from the binocular tube.
• Remove both eyepieces (
300HFig. 3-4/2) from the
box and insert them into the binocular tube to
the stop.
• Instead of an eyepiece you may insert an
auxiliary microscope (
301HFig. 3-4/3) into one of the
binocular eyepiece tubes in order to observe
aperture, phase and darkfield diaphragms and
to center phase and darkfield diaphragms.
These diaphragms can be focused with the
adjustable eye lens of the auxiliary microscope.
• The auxiliary microscope (Fig. 3-4/3) or the
pinhole diaphragm (Fig. 3-4/4) can be used to
observe conoscopic images.
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Axioscope 5/7/Vario
Mounting standard componentsZEISS
Inserting the PL 10x/23 GW focusable Pol
eyepieces (444038-9900-000) with reticle into
the binocular photo tube with upright image
(425520-9100-000)
The PL 10x/23 GW foc. Pol eyepiece contains a
reticle firmly glued into it (cannot be changed),
which is of defined orientation. When changing
the interpu-pillary distance on the binocular photo
tube, the two eyepiece tubes follow this rotary
motion synchronously, so that the position of the
orientation grooves in the eyepiece tubes remains
unchanged.
• Insert the eyepiece into the binocular photo
tube, fitting the locating screw (Fig. 3-5/2) into
Fig. 3-5 Inserting the E-PL 10x/23 GW foc.
Pol eyepieces into the binocular
photo tube with upright image
the tube’s orientation groove (Fig. 3-5/1).
The PL 10x/23 GW foc. Pol eyepiece (444038-9900-000) can be combined with a PL 10x/23 GW foc.
eyepiece (444036-9000-000).
Inserting the eyepiece reticle
Eyepiece reticles (Fig. 3-6/5) can be used with
eyepieces (Fig. 3-6/3) marked with a red dot
(Fig. 3-6/R).
• Unscrew mounting stop (Fig. 3-6/6) from the
eyepiece (Fig. 3-6/3).
• Pull locking ring (Fig. 3-6/4) out.
• Insert eyepiece reticle (Fig. 3-6/5) into the
mounting stop and fix it with the locking ring.
• Screw mounting stop (Fig. 3-6/6) into the
eyepiece (Fig. 3-6/3).
Make sure that the side of the reticle (Fig. 3-6/5)
with the line pattern on it is facing the mounting
stop in the tube (Fig. 3-6/6) (the line pattern occurs
mirror-inverted from the observer’s point of view
before the insertion). After insertion into the beam
Fig. 3-6 Inserting the eyepiece reticle
path of the microscope it is readable true to side.
The slight image shift caused by the additional path through glass is taken into account on the diopter
scale by the fact that the zero point position is indicated not by the white dot (Fig. 3-6/W), but the red
dot (Fig. 3-6/R).
NOTE
The eyepiece reticles must be inserted under dust-free conditions. This should be carried out
only by ZEISS Service.
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ZEISS Mounting standard components Axioscope 5/7/Vario
Inserting reversible eyecups
The eyepieces have rubber protection rings to avoid scratches on eyeglasses. These may be replaced by
reversible eyecups as desired.
• To do this, remove the eyeglass protection rings (Fig. 3-6/2) from the eyepieces and mount the
eyecups (Fig. 3-6/1).
Sometimes the eyeglass protection rings are seated very tightly in the eyepiece groove, so you may need
a blunt object (wooden stick) to prod them off.
3.1.5 Screwing in objectives
• Lower the mechanical stage with the stage
carrier to the bottom stop or lower the stage
carrier with the dovetail guide.
Fig. 3-7 Screwing in objectives
NOTE
Always place dust protection caps over the positions in the nosepiece which are currently not
in use.
• Remove the dust protection caps (
from the appropriate nosepiece openings.
• Take the objective (
mount it on the nosepiece (
311HFig. 3-7/3) out of its box and
312HFig. 3-7/1), starting
with the smallest magnification factor (set up
clockwise).
• Instead of an objective, you may mount an
object marker (
313HFig. 3-7/4) on any position of the
nosepiece with the aid of a W0.8/M27 adapter
314HFig. 3-7/5). Don’t forget to recap the object
(
marker when not in use to prevent it from
drying out.
310HFig. 3-7/2)
NOTE
The W0.8/M27 adapter must be used with the W0.8 objective.
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Mounting standard componentsZEISS
3.1.6 Inserting and removing push-and-click (P&C) reflector modules into the reflector
insert
To install or uninstall the modules, the reflector insert (reflector turret with 4 or 6 coded positions,
reflector slider with 2 coded positions) must be removed from the upper part of the stand.
Inserting a module:
• Remove the reflector insert (
upper part of the stand (section
315HFig. 3-8/1) from the
316H3.1.7) and put
it aside with the top side facing down.
• Insert the module (
317HFig. 3-8/3) (with the top side
facing down) with the aid of its mounting
elements on its left and right (
318HFig. 3-8/4) at a
slant from the top into the lower spring clips
319HFig. 3-8/2) on the reflector socket.
(
• Press the module against the upper spring clips
of the reflector turret until it engages firmly.
Fig. 3-8 Changing the reflector module in
the reflector insert
Removing a module:
• Disengage the module from the upper spring elements by tilting it forward, then lift it off the lower
spring elements and remove the module.
• After installing or uninstalling the reflector module, don’t forget to replace the reflector insert or the
cover cap.
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When the reflector insert is removed,
ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.7 Mounting the reflector insert
Fig. 3-9 Mounting the reflector insert
• First, remove the cover cap (
inserting the Allen wrench (AF 3) into the slot
323HFig. 3-9/2). Turn the locking screw
(
counterclockwise and pull the cover cap
forward.
• Push the reflector insert (
reflector modules P&C (e.g. reflector turret with
6 coded positions) into the upper part of the
stand until it stops, hold it and tighten the
locking screw again.
NOTE
the reflected light path is automatically
blocked so that the glare will not affect
the operator.
322HFig. 3-9/1) by
324HFig. 3-9/3) with the
Fig. 3-10 Changing the mechanical stage,
75x50 R
carrier over.
3.1.8 Mounting the mechanical stage
Depending on the type, the mechanical stages can
either be stable or they can be rotated and
centered. The shift range is 75 mm in the Xdirection and 50 mm in the Y-direction.
Stages are available with a coaxial gear knob
325HFig. 3-10/7) located on the right or the left side of
(
the stage.
To assemble/disassemble a stage, remove the stage
carrier, then reattach the stage carrier with the
mounted stage to the stand (see section 3.1.15 on
page 59).
(1) Stable mechanical stage, 75x50 R
a) How to remove the stage
• Remove the four mounting screws (
on the stage carrier (
wrench (AF 3) (
• Remove the stage (
326HFig. 3-10/6) using the Allen
327HFig. 3-10/4).
328HFig. 3-10/1) from the stage
carrier. This is easier if you turn the stage and its
325HFig. 3-10/5)
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and the stage travel range may be
Axioscope 5/7/Vario
Mounting standard componentsZEISS
b) How to mount the stage
• Fit the stage (
of the stage (
• Insert the four mounting screws (
329HFig. 3-10/1) onto the stage carrier (330HFig. 3-10/6) so that the threaded holes in the bottom
331HFig. 3-10/2) are on top of the corresponding holes in the stage carrier (332HFig. 3-10/3).
333HFig. 3-10/5) from below through the holes in the stage carrier and
screw them into the bottom of the stage.
• Tighten the screws.
NOTE
After long hours of use, the stage guide may be loose
decreased. However, this is not a malfunction and can be corrected easily as described below.
c) How to adjust the X- and Y-axis knob
tension
• In the X-direction, hold the specimen holder
and move the stage guide to the left and right
until it hits the stoppers (Fig. 3-11/1, 2).
• In Y-direction, hold the upper stage and move it
to the front and rear until it hits the stoppers.
(2) Rotatable mechanical stage 75x50/240° R
NOTE
Rotatable stages can be used only with
the stage carrier for rotatable stages
(430710-9011-000).
a) How to remove the stage
• Loosen the screw cap (334HFig. 3-12/2) of the spring
pin with approx. 3 turns.
• Press the stage forward against the spring pin
(
335HFig. 3-12/1), lift the back side off the stage
carrier (
336HFig. 3-12/3) and remove it by lifting it
upward.
• Tighten the screw cap (
337HFig. 3-12/2).
Fig. 3-11 Adjusting the X- and Y-axis knob
tension
Fig. 3-12 Changing the rotatable mechanical
stage
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ZEISS Mounting standard components Axioscope 5/7/Vario
b) How to mount the stage
• Loosen the screw caps (
• Put the stage with the dovetail ring notch onto the spring pin (
338HFig. 3-12/2) of the spring box with approx. 3 turns.
339HFig. 3-12/1).
• Press the stage forward against the spring pin and lower its back side into the stage carrier
(
340HFig. 3-12/3).
• Tighten the screw caps (
341HFig. 3-12/2).
c) How to center the stage
When using objectives with a high magnification,
only one selected objective can be centered exactly
at a time.
All stages are delivered pre-centered, i.e. when the
stage is turned, the detail of the specimen in the
focus remains in the center of the image. If the
detail shifts out of the center when the stage is
turned (
342HFig. 3-13/5), then it becomes necessary to
re-center the stage.
• Loosen the clamping screws of the stage
(
343HFig. 3-13/4) and the screw cap of the stage
carriers (
344HFig. 3-13/1).
• Determine the maximal shift of the specimen
(
345HFig. 3-13/5, arrow point) to the cross line in the
eyepiece by turning the stage.
Fig. 3-13 Centering the rotatable mechanical
stage
• Shift the specimen detail towards the center of
the cross line by turning each the two centering
• After finishing the centering procedure tighten the screw caps (
screws on the stage carrier (
Allen wrench (AF 1.5) (
346HFig. 3-13/2) with an
347HFig. 3-13/3) half the
length of the arrow. Repeat the procedure if
the specimen detail shifts out of the center
again when turning the stage.
348HFig. 3-13/1) again.
The stage can be turned by 240° within a shifting range of y ≤ 27 mm. No turning of the stage is possible
outside of this range.
(3) Adjusting the drive length on the ergonomic drives
An axial shift of the coaxial knurled knobs by a maximum of 15 mm can extend the drive length of the
X and Y shift on the mechanical stages with ergonomic drives.
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Mounting standard componentsZEISS
(4) Removing and mounting the additional
sleeves
Both coaxial knurled knobs for the stages are
equipped with additional sleeves for an even more
sensitive adjustment of the specimen position.
These sleeves can be removed in those cases when
it is important to shift the specimen more quickly.
• Loosen both clamping screws (
the lower additional sleeve (
349HFig. 3-14/4) on
350HFig. 3-14/3) and
remove them by pulling them down. Then
loosen both clamping screws (
the upper additional sleeves (
351HFig. 3-14/2) on
352HFig. 3-14/1) and
remove them by pulling them down as well.
• Re-mount the additional sleeves on the coaxial
knurled knobs by following the above steps in
reverse order. Tighten the clamping screws
Fig. 3-14 Adjusting ergonometric drive
when you are done.
(5) Adjusting the friction (smoothness of operation) on both coaxial knurled knobs of the
ergonomic drives
The smoothness of ergonomic operation is factory pre-set to a medium level. The operator can change
the friction as follows:
a) How to adjust the X-direction
• Remove the additional sleeves (
353HFig. 3-14/1 and 3) from the coaxial knurled knobs by loosening the
clamping screws.
• Shift the X-coaxial knurled knob (
354HFig. 3-14/5) downward and the Y-coaxial knurled knob (355HFig. 3-14/7)
upward.
• Hold the X-coaxial knurled knob (
357HFig. 3-14/6) to the right (increased smoothness) or left (decreased smoothness) until you reach the
(
356HFig. 3-14/5) and turn the light-colored knurled ring above it
desired level.
b) How to adjust the Y-direction
• Hold the Y-coaxial knurled knob (
359HFig. 3-14/8) to the right (increased smoothness) or the left (decreased smoothness) until you reach the
(
358HFig. 3-14/7) and turn the light-colored knurled sleeve above it
desired level.
• Replace the additional sleeves and tighten the clamping screws.
NOTE
You can increase the service life of the stage by cleaning the specimen holder regularly and
avoiding particulates from the mechanical guide system of the X-motion.
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3.1.9 Mechanical stages with friction adjustment
The drive length and friction for the X- and Y-adjustment can be adjusted individually on the standard
mechanical stage (432035-9002-000) and the mechanical stage 432035-9070-000.
(1) Adjusting the drive length on the stage
drive
The drive length of the X- and Y-drive can be
adjusted by an axial shift of the coaxial knurled
knob (
360HFig. 3-15/4 and 1) within a range of approx.
15 mm.
(2) Adjusting the frictional torque of the
coaxial knurled knobs for the
X-/Y-adjustment of the mechanical stage
The frictional torque of the coaxial knurled knobs is
factory-preset to a medium level which can be
changed as follows if necessary:
a) X-drive
Fig. 3-15 Adjusting the frictional momentum
• Slide the coaxial knurled knob for the
X-adjustment (
• Take the enclosed adjustment pin (
361HFig. 3-15/4) all the way down.
362HFig. 3-15/5)
out of the coaxial knurled knob for the
Y-adjustment (
of the holes on the lower hole nut (
• Hold the coaxial knurled knob for the X-adjustment (
365HFig. 3-15/4) and turn the hole nut with the
363HFig. 3-15/1) and stick it into one
364HFig. 3-15/3).
adjustment pin clockwise (lower frictional torque: –) or counterclockwise (higher frictional torque: +)
until you reach the desired level of friction (see
366HFig. 3-15).
• The adjustment should not be more than one rotation.
b) Y-drive
• Slide the coaxial knurled knob for the Y-adjustment (
• Stick the enclosed adjustment pin (
368HFig. 3-15/5) into the hole on the upper hole nut (369HFig. 3-15/2).
367HFig. 3-15/1) all the way up.
• Hold the coaxial knurled knob for the Y-adjustment (
370HFig. 3-15/1) and turn the hole nut with the
adjustment pin clockwise (lower frictional torque: –) or counterclockwise (higher frictional torque: +)
until you reach the desired level of friction.
• The adjustment should not be more than one rotation.
• Replace the adjustment pin in the coaxial knurled knob for the Y-adjustment (
371HFig. 3-15/1).
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000), the click stop must point to the
Axioscope 5/7/Vario
Mounting standard componentsZEISS
3.1.10 Attaching the Pol rotary stage
Two rotary stage models are available. The
mounting procedure is the same for each model.
3.1.10.1 Removing the Pol rotary stage
• Loosen the screw cap (Fig. 3-16/8) of spring pin
with about 3 turns.
• Push the Pol rotary stage (Fig. 3-16/5) to the
front against the spring-loaded pin (Fig. 3-16/9),
lift it off the stage carrier (Fig. 3-16/7) at the
back and remove it upwards.
• Retighten the screw cap (Fig. 3-16/8).
3.1.10.2 Attaching the Pol rotary stage
• If necessary, loosen the screw cap (Fig. 3-16/8)
of the spring pin with about 3 turns.
• Place the Pol rotary stage with the groove of the
dovetail (stage bottom) on the spring-loaded
pin (Fig. 3-16/9).
Fig. 3-16 Changing the Pol rotary stage with
click stop, attachable Pol object
guide and stage clips
• Attach the rotary stage (432035-9191-000) with the clamping screw (Fig. 3-16/10) pointing to the
front right.
• Push the Pol rotary stage to the front against the spring-loaded pin and lower it at the back into the
stage carrier (Fig. 3-16/7), then release it.
• Retighten the screw cap (Fig. 3-16/8).
NOTE
When you are using the rotary stage (432035-9131front.
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3.1.10.3 Removing the attachable Pol object guide and mounting stage clips
• Turn the control wheel (Fig. 3-16/3) of the Pol object guide (Fig. 3-16/1) until the clamping screw
becomes visible in the mounting hole (Fig. 3-16/2).
• Use the Allen wrench to loosen the clamping screw. Remove the Pol object guide Pol upwards.
• Insert the stage clips (Fig. 3-16/4) into the holes on the stage provided for this purpose.
• Attach the Pol object guide (453560-0000-000) for the large rotary stage (453550-0000-000) in the
same way.
• Attach the D Pol specimen holder for transmitted light specimens or the A Pol specimen holder for
reflected light specimens and mount the balance weight on the stage.
3.1.10.4 Removing the stage clips and attaching the Pol attachable object guide
• Remove stage clips (Fig. 3-16/4) from rotary stage Pol.
• Insert the Pol object guide (Fig. 3-16/1), introducing the two cylindrical pins on its underside into the
respective holes (Fig. 3-16/6) and tighten the clamping screw firmly (Fig. 3-16/2). Where required, turn
the control first to make the clamping screw accessible.
3.1.10.5 Centering the Pol rotary stage
With high-power objectives, stage centricity can be
exact only for one selected objective.
All stages are factory-pre-centered, i.e. when
rotating the stage the specimen feature set to the
center of the field of view will remain in the center.
If the specimen feature moves off the center of the
field of view (Fig. 3-17/5) while you are rotating
the stage, the stage should be re-centered as
follows.
• Before centering the stage, you must adjust the
KÖHLER illumination on the microscope (see
section 4.2.1).
• For stage centering, use a contrast-rich
Fig. 3-17 Centering the Pol rotary stage
specimen and an eyepiece with reticle.
• Loosen the stage click-stop (Fig. 3-17/4) or
clamp (on the Pol rotary stage with clamping
device) and unscrew the stage carrier cap
(Fig. 3-17/1).
• Rotate the stage to determine the position of maximum offset of the specimen feature (Fig. 3-17/5,
origin of arrow) from the center of the eyepiece reticle.
• Turn the two centering screws on the stage carrier (Fig. 3-17/2) with one Allen wrench (AF 1.5)
Fig. 3-17/3) each to move the specimen feature by half the arrow length towards the reticle center.
(
Rotate the stage again to check if the specimen feature moves off. Repeat the centering procedure, if
necessary.
• After centering, retighten the screw cap (Fig. 3-17/1).
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Mounting standard componentsZEISS
3.1.10.6 Centering objectives of polarizing
microscope stands
The nosepiece with 5-positions HF/DF/Pol and
1-position HF/DF/DIC is equipped with five
centerable objective mounts (without DIC slots) as
well as one objective mount with DIC slot (noncenterable). Accordingly, all objectives can be
centered relative to the rotary stage.
Stage centering is necessary to ensure that a
specimen feature located in the center of the field
of view does not move out while you are rotating
the stage. Centering all objectives ensures that the
specimen feature remains in the center of the field
of view even when the objective is changed.
• Before centering the stage, you must adjust the
KÖHLER illumination on the microscope (see
section 4.2.1).
Fig. 3-18 Centering objectives
• For stage centering, use a contrast-rich
specimen and an eyepiece with reticle.
• First, turn the nosepiece to swing the non-centerable objective mount (mount with DIC slot) into the
light path. Center the rotary stage for the non-centerable objective mount as described
under 3.1.10.5.
• Turn the nosepiece to move a centerable objective mount into the light path.
• Rotate the stage to determine the position of maximum offset of the specimen feature (Fig. 3-18/3,
origin of arrow) from the center of the eyepiece reticle.
• Turn the two centering screws on the nosepiece (Fig. 3-18/1) using one AF 1.5 Allen wrench
(Fig. 3-18/2) each to move the specimen feature by half the arrow length towards the reticle center.
Rotate the stage again to check whether the specimen feature moves off. Repeat the centering
procedure, if necessary.
• Center the other four objectives in the same manner.
NOTE
To preserve this centered state it is imperative to change the objectives only by turning the
nosepiece by its knurled ring, without touching the objectives.
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stage
ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.11 Mounting/removing the 80x60
motorized mechanical stage on the
Axioscope 7 motorized material
stand
The motorized stage can only be mounted
on/removed from the stand as an entire unit with
the stage carrier (see section 3.1.15 on page 59).
How to remove the stage
• Unplug the power cable connector (Fig. 3-20/1)
of the stage from the socket on the stand
(Fig. 3-20/2).
• Turn the stage (Fig. 3-19/1) and stage carrier
326HFig. 3-19/6) upside down and remove the four
(
mounting screws (Fig. 3-19/5) with the Allen
327HFig. 3-19/4).
Fig. 3-19 Changing the motorized mechanical
wrench (AF 3) (
• Carefully remove the stage from the carrier.
How to mount the stage
Fig. 3-20 Plugging in the power cable of the motorized mechanical stage
• Turn stage (329HFig. 3-19/1) and stage carrier (330HFig. 3-19/6) upside down carefully and match the threaded
holes in the bottom of the stage (Fig. 3-19/2) to the corresponding holes in the carrier (
• Insert the four mounting screws (
333HFig. 3-19/5) into the holes.
332HFig. 3-19/3).
• Align the stage in XY-direction and tighten the screws with the Allen wrench (AF 3).
• Plug the power cable connectors (
stand (
330HFig. 3-20/2).
330HFig. 3-20/1) into the sockets on the stage (330HFig. 3-20/3) and on the
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Mounting standard componentsZEISS
3.1.12 Mounting the condenser carrier
• Adjust the guide of the condenser carrier
391HFig. 3-21/3) using the coaxial knurled knob
(
392HFig. 3-21/2) until both screws (393HFig. 3-21/4)
(
become accessible.
• Mount the condenser carrier on the stage
carrier (
394HFig. 3-21/1), slide it firmly and straight
up to the upper stop and tighten both screws.
3.1.13 Mounting the condenser
• Slide the stage carrier with the focusing drive to
the upper stop.
• Unscrew both centering screws (
396HFig. 3-22/5) on
the condenser carrier until you cannot see the
end parts anymore.
• Unscrew the clamping screw (
398HFig. 3-22/9) on
the right side of the condenser carrier until the
maximum vertical adjusting range is usable.
• Slide the condenser carrier (
397HFig. 3-22/3) with the
knurled knob for vertical adjustment
398HFig. 3-22/2) all the way down.
(
• Open the front lens (if switchable) on the
condenser using the lever (
395HFig. 3-22/7).
Fig. 3-21 Mounting the condenser carrier
• Insert the condenser (
condenser carrier (
carrier (
401HFig. 3-22/1). Adjust the guide pin on the
bottom of the condenser in the direction of the
402HFig. 3-22/6).
slot (
• Push the condenser with the dovetail ring against the spring box (
399HFig. 3-22/8) between the
400HFig. 3-22/3) and the stage
Fig. 3-22 Mounting the condenser
403HFig. 3-22/4) of the condenser carrier
until the condenser can be placed on the condenser carrier horizontally.
• Release the condenser and its pin screw will slide into the front slot (
404HFig. 3-22/6).
• Tighten the centering screws until they engage in the condenser’s dovetail ring.
• Unscrew the clamping screw (
398HFig. 3-22/9) without clamping the vertical drive.
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the condenser
required to place darkfield condensers
ZEISS Mounting standard components Axioscope 5/7/Vario
NOTE
Follow the same procedure when mounting other condensers.
NOTE
If an additional component, e.g. a polarizer, has been mounted beneath
carrier, the stage carrier should be removed.
NOTE
If the Abbe condenser is used in combination with a rotary stage, ensure that the position of
the condenser head is centered as exactly as possible relative to the stage opening.
3.1.14 Mounting the darkfield condenser
Fig. 3-23 Mounting the darkfield condenser
NOTE
The condenser holder Z for darkfield is
in the condenser carrier.
• Insert the darkfield condenser (Fig. 3-23/6) in
the condenser holder Z (Fig. 3-23/5) and screw
on the fastening ring (Fig. 3-23/7).
• Push the condenser holder Z (Fig. 3-23/5)
against the spring box (Fig. 3-23/2) into the
condenser carrier (Fig. 3-23/1).
• Tighten both centering screws (Fig. 3-23/3) on
the condenser carrier (Fig. 3-23/1) until they
grip the dovetail ring and hold the condenser
holder Z (Fig. 3-23/5) in place.
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Axioscope 5/7/Vario
Mounting standard componentsZEISS
3.1.15 Mounting the stage carrier
• Screw the shoulder screw (
405HFig. 3-24/3) into the
appropriate opening:
− Opening with mark 0 (
406HFig. 3-24/3):
No sample space extension is mounted.
− Opening with mark 30 (
407HFig. 3-24/2):
The sample space extension for 30 mm is
mounted.
− Opening with mark 60 (
408HFig. 3-24/1):
The sample space extension for 60 mm
(430030-9111-000) is mounted.
• Insert the stage carrier (
409HFig. 3-24/5) at a slight
angle (beneath the shoulder screw) from the
left into the guide, then push it in straight and
tighten the wing screw (
410HFig. 3-24/4) slightly.
• Push the stage carrier along the guide upward
until it engages in the shoulder screw. Tighten
the wing screw.
• Check to ensure that the guide is accurately
positioned.
Fig. 3-24 Mounting the stage carrier
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ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.16 Changing the 12 V, 50 W halogen
lamp of the HAL 50 halogen
illuminator
• Switch off the microscope and disconnect it
from the power supply. Wait at least 15
minutes for the halogen lamp to cool down.
• Remove the HAL 50 halogen illuminator
(
411HFig. 3-25/2) from the back of the stand
412HFig. 3-25/1) and put it down with the open side
(
facing up.
• Remove the used 12 V, 50 W halogen lamp (if
Fig. 3-25 Removing the HAL 50 halogen
illuminator from the stand
necessary) by pulling it out of the cap
(
413HFig. 3-26/2) in an upward motion.
• Push the new lamp (
414HFig. 3-26/1) with its two
cap pins gently and carefully into the lamp cap
415HFig. 3-26/2). Be careful not to touch the
(
protective glass of the lamp. To avoid bending
its cap pins or damaging the lamp holder, the
lamp must not be tilted.
Fig. 3-26 Inserting halogen lamp 12 V, 50 W
• Place the HAL 50 halogen illuminator with the
connecting pins into the back side of the
microscope and push until the lamp engages
securely.
• Reconnect the microscope to the power supply.
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Mounting standard componentsZEISS
3.1.17 Mounting the illuminator TL LED10 CRI90/RL LED10 CRI90
• Using the Allen wrench (AF 3) (Fig. 3-27/7), loosen the clamping screw (Fig. 3-27/6) of the TL
illumination connector (Fig. 3-27/1) or the RL illumination connector (Fig. 3-27/2) at the lower back of
the stand.
• Connect the illuminator’s power plug (Fig. 3-27/4) to the socket (Fig. 3-27/3) on the lower part of the
stand (or the upper part of the stand).
• Match the illuminator’s (
475HFig. 3-33/5) dovetail ring to the connection piece on the stand. Ensure that
the illumination cable is not being pinched or clamped.
• Tighten the clamping screw (
477HFig. 3-33/6 or 2) with the Allen wrench (AF 3).
Fig. 3-27 Mounting the TL LED10 illuminator or RL LED10 illuminator
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ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.18 Mounting the Axioscope base plate on the stand
NOTE
When using the HAL or HBO illuminators, a base plate for Axioscope (000000-2202-526) is
mandatory.
Fig. 3-28 Mounting the base plate
• Remove the four rubber feet (Fig. 3-28/1) from the bottom of the stand.
• Align the base plate (Fig. 3-28/2) with the stand and insert a hexagonal socket screw (M6) (Fig. 3-28/3)
into each of the four holes in the base plate.
• Tighten the screws (M6) with an Allen wrench to fix the base plate to the stand.
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on the halogen
, remove the tools
the toggle
for reflected light (see
Axioscope 5/7/Vario
Mounting standard componentsZEISS
3.1.19 Mounting and adjusting the
HAL 100 halogen illuminator
The HAL 100 halogen illuminator may serve as a
light source for transmitted light and reflected light
procedures (except fluorescence light) as long as
either the lower or upper part of the stand is
equipped with the appropriate connection piece.
When mounting the halogen illuminator, no
difference is made between reflected light and
transmitted light procedures.
3.1.19.1 Mounting the HAL 100
ATTENTION
Before switching
illuminator
(437HFig. 3-31/7) included in the illuminator
case to prevent them from being
damaged by heat.
• With the Allen wrench (AF 3) loosen the
clamping screw of the respective illumination
connector (lower for TL (
420HFig. 3-29/1)) on the back of the stand.
RL (
419HFig. 3-29/8), upper for
Fig. 3-29 Mounting the HAL 100
• Insert the lamp case (Fig. 3-29/7) with the dovetail ring into the connector (
Now fixate it by tightening the clamping screw (Fig. 3-29/1 or
NOTE
422HFig. 3-29/8) with the Allen wrench.
419HFig. 3-29/2 or 420HFig. 3-29/3).
Due to the lack of ground clearance it is important to avoid tilting the illuminator when
inserting it into the lower stand part. Check for straight and correct fit.
• Connect the 3-pin lamp plug (Fig. 3-29/6) to the respective socket on the back of the external power
supply unit (suited for two lamps) (Fig. 3-29/5): plug the TL illuminator into the lower TL socket, plug
the RL illuminator into the upper RL socket.
•Connect the cable (Fig. 3-29/4) for illumination intensity control (see section 3.3.2).
NOTE
If this cable is connected to the stand, the illumination intensity of the HAL 100 in use can be
varied by rotating the Intensity/LM knob (Fig. 2-2/6).
If this cable is not connected to the stand, the illumination intensity of the HAL 100 in use can
be varied by rotating the adjusting wheel of the external power supply unit.
To switch between the illuminators for reflected and for transmitted light, flip
switch of the power supply unit to TL for transmitted light or to RL
section 3.3).
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ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.19.2 Adjusting the HAL 100
Coarse setting
Fig. 3-30 Adjusting the HAL 100
• Adjust the adjustment screws (
the gaps of the reflected image (
• After loosening the clamping screw (
429HFig. 3-29/9), remove the ready-to-operate
or
halogen illuminator (
430HFig. 3-30/2) from the
428HFig. 3-29/1
microscope stand.
• Switch on the external power supply unit of the
HAL 100 (see section 3.3.3)
431H.
• Direct the light beam towards a projecting
surface (wall) which is at least 3 m away.
CAUTION
Never look directly into the lightemitting aperture of the illuminator!
• Adjust the adjustment screw (432HFig. 3-30/3) using
an Allen wrench (AF 3) until both images of the
lamp filament appear as sharp as possible on
the projection surface.
433HFig. 3-30/4 and 5) so that the lamp filament of one image exactly fills
434HFig. 3-30/1).
Fine setting
• Mount the illuminator on the microscope stand again and tighten the clamping screw to lock it.
• If applicable, remove the filter from the beam path.
• Focus the objective ≤ 40x on the flat specular surface of a sample and go to a free area of the
specimen.
• Remove the eyepiece and center the lamp filament and its reflection in the eye pupil image with the
adjustment screws (
• Optimize the even illumination of the pupil image with the adjustment screws (
435HFig. 3-30/4 and 5).
436HFig. 3-30/3).
• If applicable, replace the filter in the beam path.
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It is not necessary to remove the
case from the stand when
) included in the
. The replacement lamp
Axioscope 5/7/Vario
Mounting standard componentsZEISS
3.1.19.3 Changing the halogen lamp 12 V,
100 W
CAUTION
Hot surface!
Danger of burns!
Always let the illuminator case cool off
for at least 15 min before touching it!
NOTE
illuminator
replacing the halogen lamp.
• Switch off the external power supply unit of the
HAL 100 (see section 439H3.3.3). Disconnect the 3pin plug from the HAL 100.
• Press the release button (
halogen illuminator HAL 100 (
the lamp carrier (
442HFig. 3-31/2) out completely
440HFig. 3-31/3) of the
441HFig. 3-31/1). Pull
and put it aside.
• Press both spring levers downwards
(
443HFig. 3-31/5) and pull out the old halogen lamp
444HFig. 3-31/6) in an upward motion.
(
• Press both spring levers downwards, insert the
new lamp into the lamp cap (
445HFig. 3-31/4) and
release the spring levers. Use only the included
special tools (
grabbing the halogen lamp. Traces of skin
grease on the halogen lamp can impair its life
446HFig. 3-31/7) when holding/
Fig. 3-31 Changing the HAL 100
span.
• Press the spring levers down again briefly. The lamp is now centered.
• Replace the lamp carrier and slide it in until it engages fully.
• Re-connect the 3-pin plug of HAL 100.
ATTENTION
Before switching on the halogen illuminator, remove the tools (437HFig. 3-31/7
illuminator case to prevent them from being damaged by heat
(438HFig. 3-31/8) may remain in the illuminator case.
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Please read the manual included with the unit before inserting or replacing the HBO 103 W/2
ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.20 Inserting the adjustment tool of
HBO 100 into the stand Bio-TL/RL
• Remove the cover (
mounting aperture of the adjustment tool.
• Insert the adjustment tool (
tighten the three included screws (
• Attach the cap to the opening (
Make sure it locks.
• Slide in the movable connecting piece of the
adjustment tool.
NOTE
Fig. 3-32 Inserting the adjustment tool
The adjustment tool is used to adjust
the HBO 100 illuminator; see
section 3.1.21.3.
3.1.21 Mounting and adjusting the HBO 100 illuminator
447HFig. 3-32/1) from the
448HFig. 3-32/4) and
449HFig. 3-32/3).
450HFig. 3-32/2).
3.1.21.1 Inserting the HBO 103 W/2 mercury vapor short arc lamp
For safety reasons, the HBO 100 illuminator and the HBO 103 W/2 mercury vapor short arc lamp are
packed separately.
NOTE
lamp.
• Insert the HBO 103 W/2 lamp into the illuminator as described in the accompanying manual.
ATTENTION
Please use an FL attenuator (if it is not installed continuously) to modify the transmission. Gray
filters are not resistant enough on the long run.
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Axioscope 5/7/Vario
Mounting standard components ZEISS
3.1.21.2 Mounting the HBO 100
illuminator
• Remove the cover from the reflected light
connection piece (
474HFig. 3-33/1).
• Insert the illuminator case (475HFig. 3-33/3) with the
dovetail ring in the reflected light connection
piece (
Tighten the clamping screw (
476HFig. 3-33/1) on the back of the unit.
477HFig. 3-33/2) with
the Allen wrench (AF 3).
• Connect the HBO 100 lamp cable to the power
supply unit (
478HFig. 3-34/1). Secure with retaining
ring.
• First, plug the cord into the outlet (479HFig. 3-34/2)
of the power supply unit, then insert the power
plug into the electrical outlet.
Fig. 3-33 Mounting the HBO 100
Fig. 3-34 Power supply unit for HBO 100
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If the FL attenuator is in the reflected
focal points of the HBO 103 W/2 lamp should be close to each other in the
ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.21.3 Adjusting the HBO 100
illuminator
The HBO 100 illuminator is available in two
versions (manual and automatic adjustment).
The self-adjusting HBO 100 (423011-9901-000)
adjusts automatically when the power supply unit
is turned on.
Fig. 3-35 Adjustment tool
Follow the instructions below to adjust the
HBO 100 illuminator manually (423010-0000-000).
NOTE
light beam path, make sure that it is set
to 100% transmission.
• Turn on the HBO 100 illuminator (480HFig. 3-36/1)
on the power supply unit for the HBO 100
481HFig. 3-55/1) and wait until it reaches its
(
operational temperature.
Fig. 3-36 Adjusting the HBO 100
• Use the adjustment screws (
for the focal point image (
NOTE
484HFig. 3-36/2 and 3) to focus the darker focal point (reflection) as you would
485HFig. 3-35/2) in the inscribed adjustment circle.
The two
adjustment circle of the adjustment tool!
• Replace the adjustment tool in its original position.
• Pull the adjustment tool (
482HFig. 3-35/1) out of the
microscope stand. The lighter colored focal
point of the lamp HBO 103 W/2 and its slightly
darker reflection become visible in the black
glass window of the adjustment tool.
• Use the knurled button (
483HFig. 3-36/4) for
collector adjustment to focus the brighter focal
point.
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Axioscope 5/7/Vario
Mounting standard componentsZEISS
3.1.22 Mounting the Colibri 3 illumination system and changing the LED modules
3.1.22.1 Mounting the Colibri 3 illumination system on the upper part of the Axioscope
stand
CAUTION
Before removing the Colibri 3 from the microscope stand, please make sure the main power is
switched off. Never look directly into the light-emitting aperture of the lamp.
• With the Allen wrench (AF 3) (Fig. 3-37/6) loosen the clamping screw (Fig. 3-37/5) of the RL
illumination connector on the upper back of the stand.
• Align the illuminator’s (Fig. 3-37/1) dovetail ring with the connection piece (
• Tighten the clamping screw (Fig. 3-37/5) with the Allen wrench (AF 3).
• Connect the illuminator plug (Fig. 3-37/4) to the stand socket (Fig. 3-37/3).
• Tighten the fixing screws on the plug.
475HFig. 3-37/2) at the stand.
Fig. 3-37 Mounting the illumination system Colibri 3
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ZEISS Mounting standard components Axioscope 5/7/Vario
3.1.22.2 Changing the LED modules in the Colibri 3 illumination system
How to change an LED module, i.e. the LED module for LED slot 1 (Fig. 3-38
/):
• Remove the Colibri 3 from the stand: Follow the instructions of section 3.1.22.1 in reverse order.
• Loosen the four captive clamping screws (Fig. 3-38/5) on the front side with an Allen wrench (AF3).
• Remove the back cover (Fig. 3-38/7).
• Disconnect the LED module power cable connector (Fig. 3-38/1.1) from the PCBA (Fig. 3-38/1.6).
• Loosen the knurled knob (Fig. 3-38/1.3) by hand and remove the old LED module (Fig. 3-38/1.4).
• Select the right LED module with matched LED-specific labels (Fig. 3-38/1.2 and 1.5) and insert it in
the correct slot (Fig. 3-38
NOTE
/
.
)
One label on the LED slot (Fig. 3-38/1.5) indicates the wavelength range for this position; the
other label on the LED module housing (Fig. 3-38/1.2) indicates the wavelength of the LED
module. See the table below as well.
• Tighten the knurled knob (Fig. 3-38/1.3) again.
• Push the LED module power cable connector (Fig. 3-38/1.1) back to the PCBA (Fig. 3-38/1.6).
• If required, replace the LED modules of LED slots 2, 3 and 4 (Fig. 3-38
/, and ) in the same
way.
• Mount the back cover (Fig. 3-38/7) and fasten the four anti-loss clamping screws (Fig. 3-38/5) with an
Allen wrench (AF3).
• Re-mount the Colibri 3 on the stand again (see section 3.1.22.1).
Fig. 3-38 Changing the LED modules in the Colibri 3 illumination system
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Axioscope 5/7/Vario
Mounting standard componentsZEISS
Usability of LED modules for Colibri 3
Position
Slot 1
Slot 2
Slot 3
Slot 4
O Usable
X Not usable
Wavelength
range
450~480 nm X O X X X
350~415 nm O X X X X
594~660 nm X X X X O
508~565 nm X X O O X
385 nm LED
423052-9593-000
470 nm LED
423052-9573-000
505 nm LED
423052-9562-000
565 nm LED
423052-9602-000
625 nm LED
423052-9522-000
3.1.23 Mounting the external illumination fixture HXP 120
Please refer to the separate operation manual of HXP 120 for instructions on installing and operating this
system.
Always disconnect the stand and the external power supply units
before beginning any mounting work.
3.2.1 Mounting the intermediate plate
for Analyzer Slider
Fig. 3-39 Mounting the binocular tube with
intermediate plate
• Loosen the screw (
wrench (AF 3) and lift off the tube (
512HFig. 3-39/5) with the Allen
513HFig. 3-39/1).
• Use the included ring tool to unscrew the tube
lens (accessible from the bottom) and keep it
free of dust.
• Screw the tube lens included with the
intermediate plate into the tube.
• Insert the intermediate plate (
514HFig. 3-39/2) with
the dovetail ring into the stand socket
515HFig. 3-39/6), adjust it, then fasten it with the
(
Allen screw (
516HFig. 3-39/4).
• Insert the analyzer slider into the slider slot
(
517HFig. 3-39/3) if applicable.
• Slightly incline the binocular tube (
when inserting it into the intermediate plate,
adjust it and fasten it with the Allen screw
(
519HFig. 3-39/4).
518HFig. 3-39/1)
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Axioscope 5/7/Vario
Mounting optional componentsZEISS
3.2.2 Mounting the tube lens turret
Mounting the tube lens turret is done similarly to mounting the intermediate plate for analyzer sliders.
See section 3.2.1.
NOTE
Before using the tube lens turret, please remove the tube lens from the used tube.
3.2.3 Mounting the magnification
changer
• Remove the camera adapter or the dust
protection cover from the camera port
521HFig. 3-40/2) of the photo tube.
(
• Mount the magnification changer (
522HFig. 3-40/1)
on the camera port, adjust it and fasten it with
the retainer nut (
523HFig. 3-40/3).
• Mount the camera with the appropriate adapter
on the camera port of the magnification
changer.
The locking power for the magnification modules
stop position can be adjusted with the screw
located on the bottom of the magnification
changer housing.
3.2.4 Changing the filters in the reflector
module FL P&C
• Remove the reflector module FL P&C
524HFig. 3-41/3) from the reflector turret (see also
(
section
525H3.1.6).
Fig. 3-41 Changing the filters in the reflector
module FL P&C
NOTE
When LED modules (423052-9593-000 for 385 nm, 423052-9573-000 for 470 nm, 4230529562-000 for 505 nm, 423052-9602-000 for 565 nm and 423052-9522-000 for 625 nm) are
used for fluorescence, the respective excitation filter is pre-installed in the LED module instead
of the reflector module.
Fig. 3-42 Mounting the filters and the color
splitter
The reflective coating (
537HFig. 3-42/4) of the color splitter (538HFig. 3-42/3) faces down when the splitter is
mounted.
The arrows (
539HFig. 3-42/7) in the figure mark the path of the illumination beam or the imaging beam.
• Unscrew the adapter ring (
526HFig. 3-41/1) with the
mounting plate included with the tools
527HFig. 3-41/6).
(
• Turn over the reflector module and let the filter
528HFig. 3-41/2 or 5) drop onto a soft surface.
(
• Insert the emission filter (
excitation filter (
position shown in
530HFig. 3-41/5) respectively at the
530HFig. 3-41.
529HFig. 3-41/2) or the
• Fix the filters with the corresponding adapter
531HFig. 3-41/1).
ring (
The emission filter and excitation filter usually have
a name and an arrow on their rims. The arrow
indicates the direction of insertion and must always
point to the inside of the reflector module (see
arrows in
532HFig. 3-41). To minimize the image offset
in multiple fluorescence images, the emission filter
may have another mark to level the point angle.
When mounting the appropriate emission filter in
the according reflector module, the mark should
be aligned with the orientation groove
(
533HFig. 3-41/4). This guarantees that the point angle
in the reflector modules acts equally and
compensates or minimizes the image offset –
already low in ZEISS filter sets – even further.
When mounting a filter without any orientation
marks (arrow), we recommend proceeding as
follows:
Filters with reflective, di-electrical coatings are
mounted with the reflective coating (
534HFig. 3-42/6)
facing outward (in relation to the reflector module)
in excitation filters (
in emission filters (
535HFig. 3-42/5) and facing inward
536HFig. 3-42/2).
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Mounting optional componentsZEISS
3.2.5 Changing the color splitter in the reflector module FL P&C
ATTENTION
Be extremely careful when installing the filters and the color splitter to avoid damaging and
soiling the optical components.
We recommend ordering completely equipped
reflector modules FL P&C, as changing the color
splitter is very challenging.
When changing the color filter, please proceed as
follows:
• Remove the reflector module FL P&C from the
reflector turret (see also section
540H3.1.6).
• Loosen both slit screws (
541HFig. 3-43/1) with a
screwdriver.
• Hold both parts of the reflector module (the emission part (
543HFig. 3-43/3) together, turn them against
part (
the mounting position and put them down.
• Now tilt the excitation part lying on top
(
544HFig. 3-44/1) upward and lift the emission part
545HFig. 3-44/4) from the holding fixtures
(
546HFig. 3-44/5b).
(
• Remove color splitter (
548HFig. 3-44/3) from the bottom part of the
box (
542HFig. 3-43/2) and the excitation
547HFig. 3-44/2) and spring
Fig. 3-43 Opening the module
module.
• Remove the old color splitter and carefully put
the new splitter with the reflective coating
facing upwards onto the spring box
(
549HFig. 3-44/3). Put both pieces together into the
bottom part of the module. Make sure that the
side latch of the spring box is in the appropriate
recess of the bottom part of the module.
• Remove the condenser (
condenser carrier (see section
580HFig. 3-48/1) from the
581H3.1.13).
If the condenser cannot be lowered sufficiently,
e.g. with mounted low-power system, then it
might become necessary to remove it with the
stage carrier, then lower it to the stop and
remove the condenser.
• Loosen the clamping screw (
condenser’s dial segment (
582HFig. 3-48/5) of the
583HFig. 3-48/3) with the
Allen wrench (AF 3) and pull out the dial
segment.
• Slide the modulator disk (
584HFig. 3-48/4) with its
two-pronged forked opening pointing forward
into the condenser. Make sure that the disk
engages in the guide on both inner sides of the
condenser. The guide serves as a stop for the
modulator disk. The pin of the disk’s clamping
screw must slide into the orientation groove of
the condenser.
• Tighten the disk’s clamping screw with the
Allen wrench (AF 3).
• Replace the condenser into its carrier (see
section
585H3.1.13).
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Mounting optional componentsZEISS
3.2.9 Changing the PlasDIC diaphragm
How to mount the slit diaphragm (426715-9000-
000):
• Turn the PhC diaphragm that needs to be
replaced into the free aperture of the modulator
disk.
• Screw the modulator disk’s centering screws
(
586HFig. 3-49/5) with the aid of an Allen wrench
back until they stop.
• Unscrew the adapter ring (
587HFig. 3-49/2) of the
PhC diaphragm with the included tool
588HFig. 3-49/1).
(
• Remove the PhC diaphragm (
spring (
590HFig. 3-49/3).
• With the aid of the tool (
slitdiaphragm (
the orientation grooves (
592HFig. 3-49/7) with the cams into
593HFig. 3-49/6).
• Screw in the adapter ring (
589HFig. 3-49/4) and
591HFig. 3-49/1), insert the
594HFig. 3-49/8) included
with the slit diaphragm with the aid of the
appropriate tool (
595HFig. 3-49/1).
3.2.10 Changing the PhC-DIC-PlasDIC
diaphragm on the
0.9 BF DF PhC DIC achromaticaplanatic condenser
Fig. 3-49 Mounting the slit-diaphragm
The diaphragm is changed according to the
directions given in sections
596H3.2.8 and 3.2.9.
ATTENTION
After inserting the DIC module, do not
tighten the condenser’s centering screw
Fig. 3-50 Changing the
PhC-DIC-PlasDIC diaphragm
again.
How to deactivate the pre-set diaphragm mechanism which automatically opens the diaphragm when
phase contrast occurs:
• Remove the cover (
597HFig. 3-50/1). Turn the screw (598HFig. 3-49/5) with the Allen wrench (AF 1.5) counter-
clockwise to the stop. You can now close the aperture diaphragm for contrasting procedures in DIC.
3.2.11 Changing the filter in the filter
wheel transmitted light
Fig. 3-51 Changing the filter in the filter
wheel transmitted light
• Remove the stage carrier (see section
• Unscrew three screws (
diaphragm socket (
600HFig. 3-51/3) of the field
601HFig. 3-51/4) and pull the
socket out of the stand base.
• If necessary, pull the filter to be changed
(
602HFig. 3-51/1) out of the corresponding position
of the filter wheel (
603HFig. 3-51/2).
• Put the new filter into the position.
NOTE
One filter position is always designed as
firmly installed shutter position.
• Repeat the procedure for all filter wheel
positions.
• Put in the field diaphragm socket and screw it in.
• Mount the stage carrier (see section
604H3.1.12).
599H3.1.12).
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Axioscope 5/7/Vario Connecting to power supply and … ZEISS
3.3 Connecting to the power supply and switching the microscope on/off
3.3.1 Microscope stand
• Connect the microscope’s power supply (
NOTE
486HFig. 3-52/2) with its power cord to an electrical outlet.
The microscope can adjust to power, voltage and frequency ranges of 100 to 240 V AC,
50-60 Hz. The power supply unit adjusts automatically to the available voltage.
• Switch the microscope on or off with the main switch (486HFig. 3-52/1) on the left side of the stand.
Fig. 3-52 Lower stand part for HAL 50 illumination (proceed similarly for LED illumination)
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ZEISS Connecting to power supply and … Axioscope 5/7/Vario
3.3.2 Connecting the HAL 100 halogen
illuminator for transmitted light
NOTE
When using the HAL illuminator, a base
plate for Axioscope (000000-2202-526)
is mandatory.
• Mount the HAL 100 (487HFig. 3-53/4) on the
transmitted light connector of the stand (see
also section
488H3.1.19.1).
• Connect the 3-pole lamp plug of the HAL 100
to the port for transmitted light TL (Fig. 3-54/3)
of the separate power supply unit for two
lamps (Fig. 3-53/2).
• Connect the cable for illumination intensity
Fig. 3-53 Lower stand part for HAL 100
illumination
control (Fig. 3-53/1) to the corresponding port
(Remote) on the back of the stand or to that of
the power supply unit (Fig. 3-54/4).
• Switch the toggle switch (Fig. 3-53/3) for
reflected or transmitted light to the TL position
(transmitted light).
Fig. 3-54 External power supply unit for two
lamps (front and back)
3.3.3 Connecting the HAL 100 halogen
illuminator for reflected light
• Mount the HAL 100 on the reflected light
connector of the stand (see section
492H3.1.19.1).
• Connect the 3-pole lamp plug of the HAL 100
to the port for reflected light RL (Fig. 3-54/2) of
the separate power supply unit for two lamps.
• Connect the power cord of the external power
supply unit (
494HFig. 3-54/5) to an electrical outlet.
• Switch the toggle switch for reflected or
transmitted light to the RL position (reflected
light).
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) to the stand, the illumination
Axioscope 5/7/Vario Connecting to power supply and … ZEISS
3.3.4 Switching the illumination on/off
• Turn the power switch (
497HFig. 3-52/1) of the
microscope with the lower part of the stand for
LED or HAL 50 illumination either on or off.
• Switch on the halogen illuminator for
transmitted light (lower stand part for HAL 100
illumination) and reflected light (if in use) at its
separate external power supply unit
(
498HFig. 3-54/1) (also refer to the separate
operation manual of the external power supply
unit). Depending on whether you want to use
the illuminator for reflected light or the
illuminator for transmitted light, switch the
toggle switch (Fig. 3-53/3) for reflected or
transmitted light to the TL position (for
transmitted light) or to the RL position (for
reflected light.
• Switch the HBO 100 illuminator for overhead
fluorescence light (if in use) on or off with the
power switch of the power supply unit for
HBO 100 (
499HFig. 3-55/1).
Fig. 3-55 Power supply unit for HBO 100
(front and back)
NOTE
If the control cable (Remote) is connected (see section 3.3.2
intensity of the HAL 100 illuminator for transmitted or reflected light, depending on which is
switched on, can be varied by rotating the Intensity/LM knob (
Fig. 2-2/6) on the lower part of
the stand.
NOTE
If the control cable is not connected to the stand, the illumination intensity of the HAL 100
illuminator in use can be varied by rotating the knob of the external power supply unit.
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ZEISS Using the Light Manager function Axioscope 5/7/Vario
3.4 Using the Light Manager function
When turned on, the previous setting of the Light Manager will be restored.
The LM function is enabled automatically if the default factory settings are active.
Disable/enable the LM function
To disable the LM function proceed as follows:
• Press one of the Snap buttons and the Intensity/LM knob simultaneously for at least 1.5 seconds.
The indicator light blinks GREEN / ORANGE / GREEN in sequence.
To enable the use of the LM function proceed as follows:
• Press one of the Snap buttons and the Intensity/LM knob simultaneously for at least 1.5 seconds.
The indicator light blinks GREEN / GREEN / GREEN in sequences.
Save the light intensity using the LM function
• Press the Intensity/LM knob for at least 1.5 seconds.
The currently set light intensity value will be saved. After it is saved, the active LED illuminator switches
off for 300 ms. The darkness in the eyepieces indicates the successful action of the Light Manager.
NOTE
If the Colibri 3 light source is connected, the Light Manager function is not available.
3.5 Default factory settings of the microscope
The default factory settings are:
− Light Manager enabled
− Light intensity set to initial minimum value
− all configuration stored will be cleared
To reset the microscope to default factory settings, proceed as follows:
• Press and hold down the Intensity/LM knob for 20 seconds.
While the knob is held down from 3 s to 20 s, the indicator light blinks in RED.
It blinks GREEN after 20 seconds. When the indicator stops blinking and remains GREEN, the reset to the
default factory setting is successful.
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Default setting of the microscopeZEISS
4 OPERATION
4.1 Default setting of the microscope
4.1.1 Setting the inter-pupillary distance on the binocular tube
• Adjust the individual inter-pupillary distance by
symmetrically swiveling of the two eyepiece
tubes towards each other (605HFig. 4-1).
The correct inter-pupillary distance has been set
when the observer sees only one round image
when looking into the eyepieces!
4.1.2 Setting the viewing height
• Adjust the viewing height to your individual
requirements by swiveling the eyepiece tubes
606HFig. 4-2/A) or down (607HFig. 4-2/B).
up (
The binocular ergonomic (photo) tubes (4255110000-000, 425512-0000-000, 445514-0000-000
and 425520-9050-000) are equipped with a
continuous height adjustment in the range from
44 mm to 50 mm.
The binocular piece of the ergo tube 4255209050-000 can also be pulled out horizontally by
50 mm.
Fig. 4-1 Setting the inter-pupillary distance
on the binocular tube
Fig. 4-2 Setting the viewing height on the
binocular tube
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ZEISS Default setting of the microscope Axioscope 5/7/Vario
4.1.3 Adjusting for ametropia (user's visual impairment) when using eyepiece reticles
The prerequisite for the correct use of an eyepiece reticle is two adjustable eyepieces (e.g. PL 10x/23 GW
foc.) to compensate for different degrees of ametropia of the user.
To adjust the eyepieces for different degrees of ametropia, follow the subsequent steps:
• Focus on the line figure of the eyepiece reticle with the focusable eye lens of the adjustable eyepiece.
• Focus on the microscopic image of a loaded specimen with the focusing drive while observing with
the eyepiece containing the eyepiece reticle.
• When both the microscopic image and the eyepiece reticle are in focus, the image for the second eye
is brought into focus with the focusable eye lens of the second eyepiece.
Both microscopic images including the eyepiece reticle are thus in focus. From this point, use only the
focusing drive to focus images.
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Illumination and contrast methods in transmitted light … ZEISS
4.2 Illumination and contrast methods in transmitted light microscopy
4.2.1 Setting up transmitted light brightfield microscopy using the KÖHLER method
(1) General principle of operation
Of all the optical microscopy methods, transmitted light brightfield microscopy is the one most commonly
used. High-contrast or tinted (stained) samples (e.g. a blood smear) can be examined easily and quickly.
For an imaging result which is as true to the specimen as possible, we must not only consider the socalled direct bundled beams but the indirect ones as well, i.e. the beams which diffract and scatter on the
sample details. According to ABBE, the image is truer to the specimen when the fraction of the cone of light is as large as possible.
The best performance of the microscope, and especially its objective, is achieved when the condenser,
field diaphragm and aperture diaphragm are adjusted in accordance with the KÖHLER illumination
principles. These fundamental basic rules for adjusting a microscope are explained in detail in
section
(2) Instrumentation for transmitted light brightfield microscopy
608H4.2.1 (3) "Setting up transmitted light brightfield microscopy using the KÖHLER method".
Every microscope (except the one with the Vario stand column) is configured to work with the
transmitted light brightfield method.
All available condensers (except special condensers like darkfield condensers) can be used for the
transmitted light brightfield method.
(3) Setting up transmitted light brightfield microscopy using the KÖHLER method
− The Axioscope has been put into operation appropriately (section
609H3).
− The Axioscope is turned on.
• Adjust the image brightness using the Intensity/LM knob (
610HFig. 4-3/1) on the microscope stand.
• Put a high-contrast sample into the specimen holder of the mechanical stage.
• If condensers with a swiveling front lens are used, swivel these into the beam path with ≥ 10x
objectives and turn the knurled knob for vertical adjustment of the condenser (
612HFig. 4-4/2) to the top stop. Make sure the stop is adjusted to prevent the condenser from lifting
and
out the sample (adjusting the condenser stop, section
• On condensers with revolver/modulator disks: turn the knurled ring (
613H4.2.1 (4)).
614HFig. 4-4/3) to position H (or B =
611HFig. 4-3/3
brightfield).
• Bring the 10x objective into position on the nosepiece (
focus drive (
616HFig. 4-3/2).
615HFig. 4-3/6) and focus the sample using the
• Close the field diaphragm (
618HFig. 4-3/A).
(
617HFig. 4-3/5) enough to make it visible in field of view (even if blurred)
• Lower the condenser using the knurled knob for vertical adjustment until the edge of the field
diaphragm appears sharp (
619HFig. 4-3/B).
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Each objective changes the size of the object field and the objective aperture. The centering
ZEISS Illumination and contrast methods in transmitted light … Axioscope 5/7/Vario
• Center the field diaphragm image with both
centering screws (620HFig. 4-3/4) on the condenser
carrier (
621HFig. 4-3/C).
• Open the field diaphragm enough to make the
edge of the diaphragm disappear from the field
of view (
622HFig. 4-3/D).
• To adjust the aperture (contrast), take an
eyepiece from the tube barrel. Looking through
the barrel adjust the aperture with the adjusting
lever (
623HFig. 4-4/4) of the aperture diaphragm to
the size of approx. 2/3 - 4/5 of the diameter of
the objective exit pupil (
624HFig. 4-3/E). In most
cases this aperture gives the best contrast at
almost full resolution and is thus the best
compromise for the human eye.
• Replace the eyepiece in the tube barrel.
Fig. 4-3 Microscope adjustment in
transmitted light brightfield
microscopy
NOTE
might also vary slightly from objective to objective, so that an adjustment of field diaphragm
and aperture might become necessary for optimal results.
With objectives < 10x, the front lens of the condenser (if swivelable) must be swiveled out of
the beam path and the aperture opened fully. In order to achieve a better contrast, the field
diaphragm can be pulled closer when working with large object fields. To do this you must
reduce the opening by a certain range. Avoid closing it too much to guarantee a consistent
illumination of the field of view.
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Illumination and contrast methods in transmitted light … ZEISS
(4) Adjusting the height stop on the
condenser carrier
• Loosen the setscrew (
625HFig. 4-4/1) on the height
stop with the Allen wrench (AF 3).
• Focus the sample with the focusing drive.
• Close the field diaphragm and adjust the
condenser vertically (
626HFig. 4-4/2) until you get a
sharp image.
• Perform the vertical adjustment on the
condenser by small increments only to avoid
lifting out the sample.
• Tighten the setscrew (
627HFig. 4-4/1) on the height
stop.
(5) Adjusting the height stop on the focusing
drive
• Turn the clamping lever (
628HFig. 4-5/1) of the stop
towards the pin stop.
Fig. 4-4 Adjusting the height stop on the
condenser carrier
• Move the stage with the aid of the focusing
drive (
629HFig. 4-5/2) to the highest position
allowable without risking colliding with
specimen holder or objectives.
• Now press the clamping lever down until the
stop engages again.
Fig. 4-5 Adjusting the height stop on the
focusing drive
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ZEISS Illumination and contrast methods in transmitted light … Axioscope 5/7/Vario
4.2.2 Setting up transmitted light darkfield microscopy using the KÖHLER method
(1) General principle of operation
Uncolored biological samples, like bacteria or living cell cultures, are sometimes hard to see in transmitted
light brightfield microscopy due to their translucence. Yet it is completely different when you examine
these samples with the transmitted light darkfield method. You basically illuminate the sample with an
illumination aperture which is higher than the one of the objective you are using.
In darkfield microscopy, only the diffracted and scattered light portions which are important for the
imaging procedure get into the objective, whereas the indirect unaffected light beams are directed past
the objective. Thus a resolution of fine structures is achieved which is partially below the resolution
capacity of a light microscope. The fine structures now appear bright and incandescent on a dark
background.
(2) Instrumentation
Condenser with transmitted light stop in position D e.g.:
− Condenser 0.9/1.25 H with modulator disk BF, DF, PhC 1, PhC 2, PhC 3,
− Condenser, achrom.-aplan. 0.9 BF DF PhC DIC,
− Darkfield condenser with dry darkfield (465505-0000-000 applicable aperture from 0.6 – 0.75),
− Ultra-condenser (465500-0000-000 applicable aperture from 0.75 – 1).
(3) Setting up transmitted light darkfield
microscopy
• Adjust the KÖHLER illumination as for
transmitted light brightfield microscopy. Instead
of the 10x objective it is necessary to use an
objective with the highest possible aperture.
The aperture must not exceed the aperture of
the condenser being used.
• Set the revolver/modulator disks to position D
and bring the condenser front lens into position
(if applicable).
• Take the eyepiece out of the tube (or replace it
with the auxiliary microscope) and check the
centering of the darkfield diaphragm in the
objective exit pupil. If the DF central darkfield
Fig. 4-6 Centering the darkfield diaphragm
on the condenser, achromaticaplanatic 0.9 H D PhC DIC
diaphragm in the universal condenser is partially
outside or is not centered to the objective exit
pupil and the pupil does not appear consistently
dark, it is necessary to re-adjust the centering of
the darkfield diaphragm.
• If the darkfield diaphragm needs to be centered, use two Allen wrenches (AF 1.5) (
adjust the two centering screws (
631HFig. 4-6/2 and 3) until the objective exit pupil appears consistently
630HFig. 4-6/1) and
dark. Remove the Allen wrenches (AF 1.5) from the condenser after finishing the centering procedure.
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. Stop the iris aperture down at least until it reaches the border
Axioscope 5/7/Vario
Illumination and contrast methods in transmitted light … ZEISS
NOTE
Objectives with built-in iris apertures have apertures which are too high for the transmitted
light darkfield microscopy
aperture for the darkfield of the condenser in use.
Performance criterion for the darkfield method is always the darkest possible background for the field of
view.
• Replace the eyepiece in the tube.
• Lighter areas which might still be visible in the field of view are reduced when the darkfield condenser
is adjusted correctly and delicately. You now have an approximately sharp eye diaphragm image.
• Now adjust the field diaphragm diameter to the size of the field of view.
Darkfield samples need to be kept impeccably clean, more so than samples for any other method. A
fingerprint, dust or any dirt particle can have a negative effect, as they brighten the background and
reduce the contrast of the object image.
(4) Setting darkfield contrast with a dry darkfield condenser
• If necessary, swivel out the low-power system, polarizer or λ plate.
• Move the condenser carrier down until the end stop.
• Place the dry darkfield condenser in the condenser holder Z (see section 3.1.14).
• Place the condenser holder Z in the condenser carrier and center it roughly, so the condenser fits into
the opening of the mechanical stage without contact, when moving upwards.
• Move the condenser up until the end stop. Place the specimen and adjust the illumination intensity to
bright enough.
• Swivel in an objective with small magnification (e.g. 5x or 10x) and focus the specimen, using the
focusing drive.
• Place a specimen as its details are distributed evenly in the field of view. This will make the image of
the field diaphragm easier to identify.
• Close the field diaphragm until the end stop.
• Lower condenser until edge of field diaphragm appears sharp (luminous-field diaphragm focus level).
There will be an increasing or decreasing light ring visible, when moving the focus upwards or
downwards from the field diaphragm focus level (so called circular "breathing" of the field diaphragm
depiction).
• Center the field diaphragm image with both centering screws on the condenser carrier.
• Swivel in the desired objective.
• Focus the specimen with the focusing drive, if necessary.
• Focus the field diaphragm, using the condenser drive. Open the field diaphragm enough to make the
edge of the diaphragm disappear from the field of view.
• Optimize the contrast with the condenser, if necessary.
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(5) Setting darkfield contrast with immersion oil darkfield condenser
• If necessary, swivel out the low-power system, polarizer or λ plate.
• Move the condenser carrier down until the end stop.
• Place the immersion oil darkfield condenser in the condenser holder Z (see section 3.1.14).
• Place the condenser holder Z in the condenser carrier and center it roughly, so the condenser fits into
the opening of the mechanical stage without contact, when moving upwards.
• Move the condenser up until the end stop.
• Place a drop of immersion oil (without bubbles, if possible) on the center of the condenser front lens.
• Place a specimen. The immersion oil will disperse between condenser and specimen holder.
• Slightly move the mechanical stage back and forth to dissipate any air bubbles in the immersion oil.
• Adjust the illumination intensity to bright enough and open the field diaphragm completely.
• Swivel in an objective with small magnification (e.g. 10x) and focus the specimen, using the focusing
drive.
• Center the field diaphragm on the condenser carrier with the adjustment screws and focus the image
with the condenser drive.
• Place a drop of immersion oil on the specimen holder, swivel in immersion oil objective and focus the
specimen.
• Close the field diaphragm until the end stop.
• Lower condenser until edge of field diaphragm appears sharp.
• Center the field diaphragm image with both centering screws on the condenser carrier.
The luminous field diaphragm appears only as a circle segment on the edge of the viewing field due to
the high magnification of the immersion oil objective. As a result, the focusing and centering of the field
diaphragm must be repeated. If necessary, the luminous-field objective should be opened slightly if the
light intensity is too low.
The field diaphragm is centered properly when the edge of the luminous field diaphragm is centered or
equidistant from the viewing field edge.
• For a sharply focused specimen, open the sharply set field diaphragm enough to make the edge of the
diaphragm disappear from the field of view.
• You can improve the contrast of the microscope image by slightly adjusting the focus level of the
condenser with the condenser drive.
• The background of the eyepiece image should appear equally dark.
• For immersion oil objectives with an iris diaphragm, the contrast can be further optimized by turning
the adjustment of the iris diaphragm.
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Illumination and contrast methods in transmitted light … ZEISS
4.2.3 Setting up transmitted light phase contrast microscopy
(1) General principle of operation
The phase contrast method is ideal for examining thin uncolored samples, e.g. culture cells. Generally,
the human eye cannot detect phase differences (variations in refractive index or thickness) within the
different cell components.
The phase contrast method uses optical modulators such as the “annular phase diaphragm and phase
ring” to convert the small phase differences in intensity and color differences which are visible to the
human eye. The interference of different beams in the intermediate image is important for the
generation of such images.
With the aid of the optically defined ring channel "annular phase diaphragm and phase ring", the highintensity direct light portions are attenuated and provided with a constant phase shift. The indirect light
portions, however, which are diffracted by different cell particles, bypass this optical channel and their
phase is affected by the difference in the sample’s refractive index and thickness.
In the intermediate image plane, the partial beams are thus differently affected and achieve interference
and strengthen or weaken each other (constructive and destructive interference) – depending on their
phase. As a result, these interferences create image contents with intensity differences visible to the
human eye.
(2) Instrumentation
− Phase contrast objectives with the phase rings PhC 1, PhC 2 or PhC 3 for various medium numeral
apertures which can also be used in the brightfield.
− Condenser with revolver/modulator disk which contains centerable ring diaphragms PhC 1, PhC 2
and PhC 3 for various medium numeral apertures.
− The phase annular diaphragm used on the condenser must be consistent with the labeling on the
objective which is used, e.g. PhC 1.
(3) Setting up transmitted light phase contrast microscopy
• Swivel the phase contrast objective, e.g. labeled Ph 1, into the beam path.
• Switch on the annular phase diaphragm on the condenser’s revolver disk which has the same label as
the phase contrast objective, e.g. 1.
• In order to check the centering and the overlap of the lighter annular diaphragm (in the condenser)
with the darker phase ring (in the objective), remove an eyepiece from the tube and replace it with an
auxiliary microscope. Use the adjusting fixture on the auxiliary microscope to focus the annular
diaphragm and the phase ring in the objective exit pupil.
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ZEISS Illumination and contrast methods in transmitted light … Axioscope 5/7/Vario
• If the overlap is not exact (632HFig. 4-7/A), the
lighter annular diaphragm must be re-centered
with the aid of two Allen wrenches (AF 1.5)
(
633HFig. 4-6/1 and 4). Adjust the two centering
screws (
634HFig. 4-6/2 and 3) so that you achieve a
full overlap with the darker phase ring
635HFig. 4-7/B).
(
• Remove the auxiliary microscope from the tube
Fig. 4-7 Centering the annular phase
diaphragm (light-colored, in the
condenser) and the phase ring
(dark-colored, in the object)
and replace it with the eyepiece.
To increase the image contrast, a green 32 x 4
interference broadband filter may be mounted on
the field diaphragm or inserted into the color glass
carrier (if available).
A complete phase contrast can only be achieved
when the light-colored annular diaphragm (in the
condenser) overlaps exactly with the dark-colored
phase ring (in the objective) in the illumination
beam path (
636HFig. 4-7/B).
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Illumination and contrast methods in transmitted light … ZEISS
4.2.4 Setting up transmitted light differential interference contrast (DIC)microscopy
(1) General principle of operation
The transmitted light DIC method allows for a high-contrast vivid display of transparent sample details.
The light is linearly polarized by a polarizer and is separated into two beams in a birefringent prism. The
two beams penetrate two parts of the sample which are closely separated. They experience different path
differences due to the different refractive indices or different sample thickness. Both beams are then
reunited in a second double refractive prism and have after passing the analyzer the same polarization.
Therefore both beams can interfere in the intermediate image and the path differences are thus
converted into intensity differences represented by a gray scale. A compensator or λ-plate may be used
for a consecutive conversion of the gray scale in a color scale.
(2) Instrumentation
− Objectives equipped with DIC fixtures, e.g. EC-Plan-Neofluar objectives
− Nosepiece with slot for DIC slider
− DIC slider, compatible with the objectives in use
− Condenser with turret disk containing DIC prisms (e.g. condenser, achromatic-aplanatic 0.9 H D
PhC DIC)
− Analyzer module ACR P&C for transmitted light in the reflector turret/slider or analyzer slider D/A
fixed or rotatable in conjunction with a mounted intermediate plate for analyzer slider 12x46
− Rotatable mechanical stage, if necessary
(3) Setting transmitted light DIC
• Put the DIC compatible objective (
638HFig. 4-8/1) into the slit of the appropriate objective position. Make sure that the slider engages firmly.
(
• Swing in the analyzer module (
637HFig. 4-8/3) in place on the nosepiece. Slide the according DIC slider
639HFig. 4-8/6) on the reflector turret (640HFig. 4-8/7) (or slide the analyzer slider
into the intermediate plate for analyzer sliders).
• Swing in the DIC prism (position DIC on the condenser) (
641HFig. 4-8/4).
• Adjust field diaphragm and aperture diaphragm according to the KÖHLER method.
• Adjust the optimal contrast on the DIC slider (
642HFig. 4-8/1) with the knurled screw (643HFig. 4-8/2).
Symmetrical adjustment of the DIC slider along its middle position lets the sample details appear as if
they were elevated or deepened.
• If desired, the compensator λ (
644HFig. 4-8/5) can be put into the opening above the nosepiece in order to
create a chromatic DIC contrast.
NOTE
The DIC method functions with polarized light. It is disturbed when birefringent elements, e.g.
foils, are put between polarizer and analyzer, as is sometimes done when doing a histological
incision. The same situation occurs with Petri dishes or specimen holders which have a plastic
base. In these cases we recommend using PlasDIC.
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ZEISS Illumination and contrast methods in transmitted light … Axioscope 5/7/Vario
1DIC slider
2Knurled screw
3Objective on the nosepiece
4Condenser with DIC prism
5Compensator λ6Analyzer module
7Reflector turret
Fig. 4-8 Components for the transmitted light DIC method
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Illumination and contrast methods in transmitted light … ZEISS
4.2.5 Setting up transmitted light PlasDIC contrast microscopy
(1) General principle of operation
PlasDIC can be used independently from the material of the specimen holder.
The contrast method gives a relief-like image and is especially well suited for thicker objects. The contrast
is adjustable. It is possible to contrast the cavities of microtiter plates up to the edge. It is not necessary to
use cultivation holders with a glass base.
(2) Instrumentation
− Abbe condenser with modulator disk and objective-dependent 2 mm slit diaphragm (A-Plan 10x
and LD A-Plan 20x) or 4.5 mm slit diaphragm (in all other cases)
− ACR P&C analyzer module for transmitted light and reflector turret /slider or D/A fixed or rotatable
analyzer slider in connection with mounted intermediate plate for analyzer slider 12x46
(3) Setting PlasDIC
• Fully open the aperture on the condenser.
• Place the sample on the stage.
• Swing the position with the 2 or 4.5 mm slit diaphragm for PlasDIC of the condenser into the beam
path. Increase brightness when switching from brightfield to PlasDIC.
• Swing the analyzer module into the beam path or slide analyzer slider in beam path in intermediate
plate.
• Swing in the objective for PlasDIC.
• Slide the PlasDIC slider on the nosepiece into the DIC slit of the objective in use.
• Adjust the contrast with the knurled screw of the PlasDIC slider. You can now see the structures in
relief or in pseudo-darkfield. The best result is achieved in the relief mode.
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4.2.6 Setting transmitted light polarization
4.2.6.1 Detecting birefringence
(1) Application
The transmitted light polarization method is used for samples which change the polarization of the light.
Such samples are called birefringent. Examples include crystals, minerals or polymers. If such birefringent
substances are observed between crossed polarizers, the birefringent portion of the sample appears
bright while its surroundings remain dark.
A birefringent substance can be recognized by turning the sample by 360° between crossed polarizers.
The sample should show four bright and four dark appearances during the turning procedure. During the
turning procedure, interference colors appear that range from gray (mostly for biological samples)
through white, yellow and red until blue, depending on birefringence, thickness as well as sample
orientation. The interference colors may be of the first or a higher order.
(2) Instrumentation
− Strain-free objectives
− Pol rotary stage
− D Polarizer (rotatable or fixed), if no polarizer is already integrated in the condenser.
− ACR P&C analyzer module for transmitted light in the reflector turret/slider or D analyzer slider
fixed or with lambda-plate.
− Depolarizer for avoiding unwanted polarizing effects
A depolarizer (quartz depolarizer) should be incorporated in all microscopes used to examine
mineralogical/geological specimens.
A depolarizer extinguishes undesirable polarization effects possibly (e.g. false or pseudo-pleochroism)
occurring behind the analyzer (e.g. on prism surfaces in the tube), or shifts them to higher orders.
• Insert the quartz depolarizer with tube lens 115 (428106-9010-000) into the tube
or
• insert the quartz depolarizer with tube lens 130 in combination with intermediate plate (428106-
9000-000) into the tube.
(3) Setting the microscope
• Adjust the microscope as described in section
645H4.2.1 (3) for transmitted light brightfield microscopy
using the KÖHLER method.
• Center the rotary stage Pol (
• Swing the polarizer (
647HFig. 4-9/3) into the beam path and position it to 0° if you are using a rotatable
646HFig. 4-9/1) (see section 3.1.10.5) and objectives (see section 3.1.10.6).
polarizer.
• Put the analyzer (
648HFig. 4-9/2) into the slit for the compensator or swing analyzer module on the
reflector turret/slider. Because of the crossed polarizers the field of view now appears dark.
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Illumination and contrast methods in transmitted light … ZEISS
Fig. 4-9 Components for transmitted light polarization
NOTE
If an analyzer slider is going to be used for working with the Axioscope, either the reflector
module H P&C or an empty position must be swiveled in on the reflector turret/slider.
• Bring the specimen to be examined into the field of view and turn it with the rotary stage. Normally,
birefringent (anisotropic) objects will now show the same color and intensity variations as described
above during rotation between crossed polarizers. Optically anisotropic substances may remain dark
when an isotropic direction, e.g. from optically single-axle or double-axle crystals, is placed parallel to
the observation direction.
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γ
ZEISS Illumination and contrast methods in transmitted light … Axioscope 5/7/Vario
4.2.6.2 Determination of the polarization direction nγ'
(1) Application
The determination of the polarization direction of n
absolute or relative largest index of refraction) and n
or nγ' respectively (polarization direction with the
γ
or nα' respectively (polarization direction with the
α
absolute or relative smallest index of refraction) relative to the morphological directions, e.g. of crystal
surfaces, crystal needles or fibers, provide an important signature of the material. This method is also
used in the diagnosis of bio-crystals (e.g. gout and pseudo-gout).
Fig. 4-10 Determination of the polarization direction n
in a synthetic fiber
'
(2) Instrumentation
− Eyepiece with reticle
− Strain-free objectives
− Rotary stage (
649HFig. 4-9/1)
− D polarizer (rotatable or fixed), if not using a condenser with an integrated polarizer
− ACR P&C analyzer module for transmitted light in the reflector turret/slider or analyzer slider D
fixed or with lambda plate
− Pol adjusting tool sample for polarization microscopy (453679-0000-000)
(3) Setting the microscope
• Adjust the microscope as described in section 4.2.1
Make sure the inter-pupillary distance is adjusted correctly on the binocular tube (see section
• Center the rotary stage Pol (
• Swivel the polarizer (
653HFig. 4-9/3) into the beam path and position it to 0° if you are using a rotatable
652HFig. 4-9/1) and objectives (see sections 3.1.10.5 and 3.1.10.6).
650H(3) for transmitted light brightfield microscopy.
651H4.1.1).
polarizer.
• Swivel in the analyzer module on the reflector turret (
654HFig. 4-9/2) (or insert the analyzer slider into the
intermediate plate). Because of the crossed polarizers the field of view now appears dark.
• Place the Pol adjusting tool sample on the microscope stage and turn until the sample appears dark.
• Remove the analyzer from the beam path and align the reticle along the split cracks of the sample.
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