Axiovert 200 & Axiovert 200 M Inverted Microscope Operating Manual

INTRODUCTION Copyright

Carl Zeiss

Axiovert 200

Knowledge of this manual is required for the operation of the instrument. Would you please therefore make yourself familiar with the contents of this manual and pay special attention to hints concerning the safe operation of the instrument. The specifications are subject to change; the manual is not covered by an update service. ©

Unless expressly authorized, forwarding and duplication of this document, and the utilization and communication of its contents are not permitted. Violations will entail an obligation to pay compensation. All rights reserved in the event of granting of patents or registration of a utility model.

Issued by:

Carl Zeiss Light Microscopy P.O.B. 4041 37030 Göttingen GERMANY Phone : ++49 551 5060 660 Telefax: ++49 551 5060 464 Internet : www.zeiss.de/micro E-Mail: [email protected]

Number of this manual: B 40-080 e Date of issue: 30.03.2001

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INTRODUCTION Contents

Axiovert 200

Carl Zeiss

CONTENTS page INTRODUCTION ... 0-2 Copyright ...0-2 Contents... 0-3 Notes on instrument safety... 0-7 Notes on warranty ... 0-10 Overall view of the Axiovert 200... 0-11 Overall view of the Axiovert 200 M... 0-11 Microscopy in transmitted-light brightfield in a few steps ... 0-12

CHAPTER 1

INSTRUMENT DESCRIPTION ... 1-3

1.1

Name and intended application ...1-3

1.2

Instrument description and main features ...1-5

1.3

Microscope configurations and modules ...1-6

1.4

Objectives ... 1-10

1.5

Eyepieces ... 1-12

1.6

Condensers... 1-12

1.7

Specimen stages and mounting frames... 1-13

1.8

Binocular tubes ... 1-13

1.9

Technical Data ... 1-14

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Carl Zeiss

INTRODUCTION Contents

Axiovert 200

page CHAPTER 2

START-UP... 2-5

2.1

Unpacking and installation of the microscope... 2-5

2.2 2.2.1 2.2.2

Attachment of binocular (photo) tube ... 2-6 Inserting the eyepieces and the centering telescope... 2-6 Inserting the eyepiece reticle ... 2-7

2.3 2.3.1 2.3.2

Attachment of transmitted-light illumination ... 2-8 Attachment of carrier for transmitted-light illumination (100 W) ... 2-8 Attachment of carrier for transmitted-light illumination (30 W) ... 2-8

2.4

Screw in objectives ... 2-9

2.5 2.5.1

2.5.4 2.5.5

Attachment of microscope stages ... 2-10 Attachment of mechanical stage 130x85 R/L and mounting frame for mechanical stage (K)... 2-10 Attachment of scanning stage... 2-10 Attachment of specimen stage 250x230, object guide and mounting frame for object guide (M) ... 2-11 Attachment of heating stage ... 2-12 Attachment of gliding stage Z ... 2-12

2.6 2.6.1 2.6.2 2.6.3

Attachment of condensers ... 2-13 Condensers for the Axiovert 200... 2-13 Condensers from the Axioplan 2 imaging / Axioskop 2 product line ... 2-13 Changing the DIC prism in the condenser turret ... 2-14

2.7 2.7.1 2.7.2 2.7.3 2.7.4

Reflector turret ... 2-15 Attachment of reflector turret... 2-15 Equipment of reflector turret ... 2-15 Changing the filter set in the FL reflector module ... 2-16 Changing the beam splitter in the FL reflector module ... 2-16

2.8

Connection to the line ... 2-18

2.9

Interfaces of the Axiovert 200 M ... 2-19

2.10

Switch microscope and ebq 100 dc power supply on and off... 2-19

2.11

Equipotential bonding terminals ... 2-19

2.12 2.12.1 2.12.2 2.12.3 2.12.4

HAL 100 halogen illuminator... 2-20 Change / attachment of the HAL 100 halogen lamp ... 2-20 Coarse alignment of halogen illuminator ... 2-21 Attachment of halogen illuminator... 2-21 Fine alignment of halogen illuminator ... 2-22

2.5.2 2.5.3

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Axiovert 200

Carl Zeiss

page 2.13 2.13.1 2.13.2 2.13.3 2.13.4

Illuminator N HBO 103 ... 2-23 Attachment / change of the HBO 103 W/2 mercury pressure short-arc lamp... 2-23 Coarse alignment of illuminator N HBO 103... 2-25 Attachment of illuminator N HBO 103 ... 2-26 Fine adjustment of illuminator N HBO 103 using the adjusting aid... 2-27

2.14

Attachment of Aqua Stop ... 2-28

CHAPTER 3

OPERATION... 3-3

3.1 3.1.1 3.1.2

Axiovert 200 (manual) ...3-4 Operation and function controls on the Axiovert 200 (manual)... 3-4 Switching on and basic settings on the Axiovert 200 (manual)... 3-14

3.2 3.2.1 3.2.2

Axiovert 200 M (motorized)... 3-16 Operation and function controls on the Axiovert 200 (motorized)... 3-16 Switching on and basic settings on the Axiovert 200 M (motorized) ... 3-23

3.3 3.3.1 3.3.2 3.3.3 3.3.4 3.3.5

Illumination and contrasting techniques... 3-28 Setting of transmitted-light brightfield for KÖHLER illumination... 3-28 Setting of transmitted-light phase contrast ... 3-33 Setting of differential interference contrast (DIC) in transmitted light... 3-35 Setting of VAREL contrast in transmitted light... 3-38 Setting of fluorescence contrast in reflected light ... 3-40

3.4 3.4.1 3.4.2 3.4.3

Documentation ... 3-43 Image orientation of camera ports... 3-43 Photomicrography with SLR camera... 3-45 Photomicrography using a digital camera and videomicroscopy ... 3-46

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Carl Zeiss

INTRODUCTION Contents

Axiovert 200

page CHAPTER 4

CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE... 4-3

4.1

Care ... 4-3

4.2 4.2.1 4.2.2 4.2.3

Maintenance ... 4-4 Performing checks ... 4-4 Changing the fuses on the microscope... 4-4 Changing the fuses on the ebq 100 dc power supply ... 4-5

4.3

Service... 4-6

ANNEX ...A-1 List of abbreviations ...A-3 List of key words...A-5 Certification in accordance with DIN ISO 9001 / EN 46001 EC Declaration of Conformity

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Axiovert 200

INTRODUCTION Notes on instrument safety

Carl Zeiss

Notes on instrument safety The Axiovert 200 / Axiovert 200 M microscopes were designed, produced and tested in compliance with DIN 61010-1 (IEC 1010-1), Safety requirements for electrical measuring, control and laboratory instruments, and meet the requirements of appendix I of directive 73/23/EC and the relevant CSA and UL directives. The instruments meet the requirements of the EC directive 89/336/EC and the EMC legislation of November 9th 1992. This operation manual includes information and warnings which must be observed by the user. The following warning and information symbols are used in this manual:

☞

NOTE This symbol is a warning which you must observe under all circumstances. CAUTION This symbol is a warning which indicates a hazard to the instrument or instrument system.

CAUTION This symbol is a warning which indicates a hazard to the user of the instrument.

CAUTION Hot surface!

CAUTION UV radiation is emitted!

CAUTION Disconnect the instrument from the line before opening it!

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Carl Zeiss

INTRODUCTION Notes on instrument safety

Axiovert 200

The Axiovert 200 microscopes, including original accessories, may only be used for the microscope techniques described in this manual. Particular attention must be paid to the following notes: The manufacturer cannot assume any liability for any other applications, possibly also involving individual modules or single parts. This also applies to all service or repair work which is not carried out by authorized service personnel. Furthermore, this forfeits all the claims against warranty The power plug must be inserted in a socket featuring a grounding (earth) contact. The grounding effect must not be made ineffective by an extension cable which does not have a protective ground wire.

If it is established that the protection measures are no longer effective, the instrument must be switched off and safeguarded against inadvertent operation. For repair of the instrument, contact the Carl Zeiss microscope service in Germany (see page 4-6) or your local Carl Zeiss agency.

The Axiovert 200 microscopes are not equipped with any special devices for protection from substances which are corrosive, toxic, radioactive or otherwise hazardous to health. All the legal regulations for accident prevention, particularly those in the respective countries, must be observed when handling such substances.

Before switching on the power unit for the HBO 50/100, check whether it is suitable for the line voltage present. Always disconnect the instrument from the line before opening the instrument and before changing the fuses. Make sure to use only fuses of the rated power required. The use of makeshift fuses and the short-circuiting of the fuse holders are not permitted.

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Axiovert 200

INTRODUCTION Notes on instrument safety

Carl Zeiss

Gas discharge lamps, e.g. HBO 100, emit ultraviolet radiation which can cause burns on the eyes and skin. Therefore, never look directly into the light of these lamps and avoid direct, unprotected incidence of their light on your skin. When using the microscope, always use the protective devices belonging to the instrument (e.g. special attenuation filters). When hot, gas discharge lamps are under high internal pressure and must therefore be changed when cooled down by using protective gloves and goggles (for detailed information, please see the operating instructions B 40-065 e).

When fluorescence filters are used, the filter protecting from heat emitted by the microscope illuminator must not be removed, since fluorescence filters are sensitive to heat and their function might therefore be impaired.

Placing objects against or covering ventilation slats can lead to a build-up of heat which will damage the instrument and, in extreme cases, cause a fire. Always keep the ventilation slats clear and make sure that no objects enter the instrument through the ventilation slats.

Avoid touching the hot lamp housings. Always pull the power plug before changing the lamps and allow the instrument to cool down for approx. 15 mins.

Dust and dirt can impair the performance of the instrument. Therefore, the instrument must be protected against these influences as far as possible, and covered with the dust cover if it is not used for longer periods of time. Always check whether the instrument is switched off before you cover it.

The instruments may only be operated by trained personnel who must be aware of the possible danger involved with microscopy and the relevant application. The Axiovert 200 microscopes are opto-mechanical precision instruments which can be impaired in their performance or damaged when handled improperly.

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Carl Zeiss

INTRODUCTION Notes on warranty

Axiovert 200

Notes on warranty The manufacturer guarantees that the instrument has no material and production defects when delivered. You must inform us of any defects immediately and we must do anything to minimize the damage. If the manufacturer is informed of such a defect, he is obliged to remove it; it is his decision whether he does this by repairing the instrument or by delivering an instrument free of any defect. No guarantee is provided for defects caused by natural wear (wearing parts in particular) and improper use. The instrument manufacturer is not liable for damage caused by faulty operation, negligence or any other meddling with the instrument, particularly the removal or replacement of instrument components, or the use of accessories from other manufacturers. This forfeits all the claims against warranty. With the exception of the work specified in this manual, no maintenance or repair of the microscopes may be performed. Repairs may only be performed by Carl Zeiss service staff or specially authorized personnel. Should any defect occur with the instrument, please get in touch with the Carl Zeiss microscopy service in Germany (see page 4-6) or your local Zeiss agency.

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Axiovert 200

B 40-080 e 03/01

INTRODUCTION Overall view of the Axiovert 200 and Axiovert 200 M

Carl Zeiss

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Carl Zeiss

INTRODUCTION Microscopy in transmitted-light brightfield in a few steps

Axiovert 200

Microscopy in transmitted-light brightfield in a few steps

☞

Before starting to use the Axiovert 200, make sure to read the notes on instrument safety and the chapters entitled "Instrument Description" (Chapter 1) and "Start-up" (Chapter 2).

• Make the microscope ready for operation as described in chapter 2 and switch it on via the On/Off switch (0-2/1). • Select the objective with the lowest magnification (e.g. 10x) on the nosepiece (0-2/2). Set factor 1x on the setting wheel (0-2/4) of the Optovar turret. • Open the luminous-field diaphragm or the aperture diaphragm completely by pulling lever (0-2/16) to the front until stop or by turning the setting wheel (0-2/20) to the front until stop. • Turn the setting ring (0-2/19) to move the condenser turret in position H for brightfield (or DIC). • Move reflector turret (0-2/5, if available) into the position without filter combination via the setting ring. • If required, remove analyzer slider (0-2/3) or switch to free light path. • Turn setting wheel for Sideport right / left / vis (0-2/22) to position 100 % vis (visual

).

• Turn setting knob for Frontport / Baseport / vis (0-2/23) to position 100 % vis ( VIS ). • Set beam splitting ratio to 100 % vis (0-2/10) on the tube. Switch off the Bertrand lens (if available). ). Move combined rotary / slider knob (0-2/9) to position 100 % vis ( • Place a high-contrast specimen on the microscope stage (0-2/21). Adjust the binocular component. • Use the coarse / fine focusing drive (0-2/6) to focus on the selected detail of the specimen. Should no light be visible in the eyepieces, switch on the halogen illuminator via the HAL on / off switch (0-2/7). • Use the toggle switch (0-2/8) to set the light intensity to comfortable brightness.

Fig. 0-1

Diaphragm settings in transmittedlight brightfield according to KÖHLER

• Close luminous-field diaphragm (0-2/16) until it is visible in the field of view, even if not in focus (0-1/A). • Focus on the edge of the luminous-field diaphragm (0-1/B) by moving the condenser (0-2/17) vertically.

• Center (0-1/C) luminous-field diaphragm via the centering screws (0-2/15 and 18) and open it until the edge of the diaphragm just disappears from the field of view (0-1/D). • Remove one eyepiece from the eyepiece tube (or swing in Bertrand lens) and set aperture diaphragm (0-2/20) to approx. 2/3 of the diameter of the objective exit pupil (0-1/E). Optimum contrast setting is dependent on the respective specimen. • Insert the eyepiece again (or swing out Bertrand lens) and refocus, if required, via the fine drive. • After the microscope has been set to transmitted-light brightfield in this way, changing to this special contrasting technique is now possible (see chapter 3 of this manual).

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Axiovert 200

INTRODUCTION Microscopy in transmitted-light brightfield in a few steps

1 On / Off switch 2 Objective nosepiece 3 Compartment for slider Analyzer 4 Setting wheel for Optovar turret 5 Reflector turret 6 Focusing drive coarse / fine 7 HAL on / off switch 8 Toggle switch for illumination intensity 9 Turning or sliding knob for vis / doc beam splitting

Fig. 0-2

10 Turning or sliding knob for Bertrand lens and manual shutter 11 Binocular tube component 12 Setting ring of the eyepiece 13 Eyepiece 14 Polarizer D with 2-position filter changer 15 Centering screw for condenser 16 Setting lever for luminous-field diaphragm 17 Setting knob for vertical adjustment of the condenser

Carl Zeiss

18 Centering screw for condenser 19 Turret disk of condenser 20 Setting wheel for aperture diaphragm on the condenser 21 Microscope stage 22 Compartment for aperture diaphragm slider 23 Setting wheel for Sideport 24 Setting knob for Frontport / Baseport

Axiovert 200

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Carl Zeiss

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INTRODUCTION Microscopy in transmitted-light brightfield in a few steps

Axiovert 200

B 40-080 e 03/01

Axiovert 200

INSTRUMENT DESCRIPTION Contents / List of illustrations

Carl Zeiss

INSTRUMENT DESCRIPTION Contents 1

INSTRUMENT DESCRIPTION ... 1-3

1.1

Name and intended application...1-3

1.2

Instrument description and main features...1-5

1.3

Microscope configurations and modules...1-6

1.4

Objectives... 1-10

1.5

Eyepieces... 1-12

1.6

Condensers ... 1-12

1.7

Specimen stages and mounting frames ... 1-13

1.8

Binocular tubes... 1-13

1.9

Technical Data ... 1-14

List of illustrations Fig. 1-1 Fig. 1-2 Fig. 1-3 Fig. 1-4

Microscope configurations and modules (sheet 1) ...1-6 Microscope configurations and modules (sheet 2) ...1-7 Microscope configurations and modules (sheet 3) ...1-8 Microscope configurations and modules (sheet 4) ...1-9

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1-1

Carl Zeiss

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INSTRUMENT DESCRIPTION Contents / List of illustrations

Axiovert 200

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Axiovert 200

INSTRUMENT DESCRIPTION Name and intended application

1

INSTRUMENT DESCRIPTION

1.1

Name and intended application

Manufacturer's name:

Inverted microscope for transmitted light and epifluorescence

Brief name:

Axiovert 200 Axiovert 200 M

Carl Zeiss

(manual version) (motorized version)

The Axiovert 200 microscopes fit in the product family of inverted transmitted-light microscopes as follows: Laboratory microscopes

Research microscopes

− Axiovert 25

− Axiovert 200

− Axiovert 25 C

− Axiovert 200 M

− Axiovert 25 CFL The Axiovert 200 microscopes are universally applicable inverted microscopes and are mainly used for the examination of cell and tissue cultures and of sediments in culture flasks, Petri dishes, microtiter plates, etc. in transmitted and reflected light. The Axiovert 200 microscopes permit the performance of the transmitted-light techniques brightfield, phase contrast, differential interference contrast and VAREL contrast, and the epi-fluorescence technique. The microscopes Axiovert 200 and Axiovert 200 M are the basis for scientific microscopic work on living cells. − The sturdy stand provides attachment possibilities for various tools (micromanipulation), different light sources, temperature control devices, etc. − Ample space is available for the specimens and the relevant handling systems on account of the inverted design, the LD illumination system of the microscope and the use of fixed stages. This permits the performance of experiments which would not be possible with upright microscopes. − The design allows the easy attachment of cameras, lasers, specific stages, etc.

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Carl Zeiss

INSTRUMENT DESCRIPTION Name and intended application

Axiovert 200

Typical fields of application: Observation of intracellular processes in living cell cultures, cell/cell interactions, motility, growth, measurements of potential, drug detection, microinjection, IVF (in-vitro fertilization), toxicity examinations, patch-clamp technique, ion measurements, digital recording, long-time / time lapse examinations in combination with the automation of processes, z-sectioning, deconvolution, visualization of molecular structures, Fura (Ca measurement), GFP, optical tweezers and scissors, single molecule detection, TIRF ... Accessories for temperature control and incubation are described in the separate manual B 40-610 d/e, "Incubation systems on the Axiovert".

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Axiovert 200

1.2

INSTRUMENT DESCRIPTION Instrument description and main features

Carl Zeiss

Instrument description and main features

The Axiovert 200 / Axiovert 200 M is available either as manual or as motorized version. The accessory components are part of a modular system. For documentation purposes, the Axiovert 200 / Axiovert 200 M can be equipped with the maximum of five camera / TV ports in accordance with the customer's requests. Adaptation possibilities are provided for heating stages, incubators and micromanipulators. Major instrument features: − ICS optics for image creation − high thermal and mechanical stability − high flexibility in documentation − improved ergonomic design − LCD display of instrument parameters − 23 mm field of view − Light Manager − modular design for optimum adaptation to the relevant application − 6-position nosepiece, coded − 5-position reflector turret, manual: can be changed on both sides; motorized: can be changed from the right − 5-position or 6-position condenser turret − 3-position Optovar turret − changeable aperture diaphragm and luminous-field diaphragm sliders in reflected light − fluorescence shutter − illuminators: HAL 100 W, HBO 50, N HBO 103, N XBO 75 − Axiovert 200 M, all the major microscope functions are motorized.

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Carl Zeiss

INSTRUMENT DESCRIPTION Microscope configurations and modules

1.3

Microscope configurations and modules

Fig. 1-1

Microscope configurations and modules (sheet 1)

1-6

Axiovert 200

B 40-080 e 03/01