Zeiss DirectFRAP User manual

DirectFRAP
Operating Manual

Carl Zeiss Copyright DirectFRAP
2 423635-9001-701 05/2014
Knowledge of this manual is required for the operation of the instrument. You should therefore
familiarize yourself with the contents of this manual and pay special attention to instructions concerning
the safe operation of the instrument.
Subject to alterations in the interest of technical progress. The manual is not covered by an update
service.
© Unless explicitly authorized, dissemination or duplication of this document and commercial
exploitation or communication of its contents are not permitted. Persons in contravention of this
copyright shall be liable to pay compensation for damages.
All rights reserved in the event of a patent grant or registration of a utility patent.
Published by: Carl Zeiss Microscopy GmbH
Carl-Zeiss-Promenade 10
07745 Jena, Germany
[e-mail]
www.zeiss.com/microscopy
Carl Zeiss Microscopy GmbH
Königsallee 9-21
37081 Göttingen, Germany
Order number: 423635-9000-701
Date of publication: Version 7, 22/05/2014

DirectFRAP Contents Carl Zeiss
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Contents
Page
1Notes on Instrument Safety ............................................................................................ 5
1.1 General Notes .................................................................................................................... 5
1.2 Safety Instructions .............................................................................................................. 7
1.2.1 Notes on Laser Radiation .................................................................................................... 7
1.2.2 Installation Notes................................................................................................................ 8
1.2.3 Notes on Computer and Software..................................................................................... 10
1.2.4 Service Interfaces.............................................................................................................. 11
1.2.5Warning and Information Labels on the Instrument ........................................................... 17
1.3 Notes on Warranty ........................................................................................................... 20
2INSTRUMENT DESCRIPTION........................................................................................... 21
2.1 Intended Use.................................................................................................................... 21
2.2 Functional Principle........................................................................................................... 22
2.3 Instrument Description and Main Features......................................................................... 23
2.3.1 Laser Manipulation Slider.................................................................................................. 24
2.3.2 Laser Bench Tower or Laser Bench Rack ............................................................................ 25
2.3.3 Signal Distribution Box...................................................................................................... 26
2.3.4 Basic Laser Safety Kit for Axio Observer.Z1 Stand............................................................... 27
2.3.5 Remote Control Switch for Laser Operation....................................................................... 28
2.4 System Overview - DirectFRAP Imaging System.................................................................. 29
2.5 Diaphragm Size and Objective Magnification..................................................................... 30
2.6 Optical Fibers for the Application of the DirectFRAP Laser Manipulation Slider .................... 31
2.7 Suitability of Lasers for the Application of the DirectFRAP Imaging System.......................... 31
2.8 Laser Safety Concept ........................................................................................................ 32
2.9 Technical Data.................................................................................................................. 33
3START-UP ....................................................................................................................... 37
3.1 Installation Requirements.................................................................................................. 37
3.1.1 Space Requirements ......................................................................................................... 37
3.1.2 Power Requirements......................................................................................................... 38
3.1.3 Ambient Conditions ......................................................................................................... 39
3.2 Requirements for the Microscope Stand ............................................................................ 40
3.3 Basic Microscope Settings ................................................................................................. 40
3.4 Connecting the Fiber Oscillator ......................................................................................... 41
3.5 Inserting the Laser Manipulation Slider .............................................................................. 42
4OPERATION .................................................................................................................... 43
4.1 Control and Functional Elements....................................................................................... 43
4.1.1 DirectFRAP Imaging System............................................................................................... 43
4.1.2 Laser Bench Tower ........................................................................................................... 44
4.1.3 Laser Bench Rack/Laser Bench Rack 3F .............................................................................. 44

Carl Zeiss Contents DirectFRAP
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4.2 Switching on.....................................................................................................................45
4.3 During Operation ..............................................................................................................45
4.4 Switching off ....................................................................................................................45
4.5 Adjusting the Laser Beam with the Laser Manipulation Slider..............................................46
4.6 Operation via ZEN Software (Blue Edition) ..........................................................................47
4.7 Experiment Procedure .......................................................................................................47
5CARE, MAINTENANCE, TROUBLESHOOTING AND SERVICE ..........................................49
5.1 Instrument Care................................................................................................................49
5.2 Instrument Maintenance ...................................................................................................49
5.3Troubleshooting................................................................................................................50
5.4 Changing the Beam Combiner ..........................................................................................51
5.5 Changing the Fuses...........................................................................................................52
5.5.1 Signal Distribution Box ......................................................................................................52
5.5.2 Laser Bench Tower ............................................................................................................52
5.5.3 Laser Bench Tower Coupling Module.................................................................................53
5.6 Mounting the Camera.......................................................................................................54
5.7 Connect HXP 120 Illumination Unit....................................................................................55
5.8 Inserting Filters in the Excitation Filter Wheel 8x mot. .........................................................56
5.9 Mounting the Excitation Filter Wheel 8x mot......................................................................57
5.10 Changing the HAL Halogen Lamp......................................................................................57
5.11 Service Request .................................................................................................................58
6APPENDIX .......................................................................................................................59
6.1 List of Abbreviations..........................................................................................................59
6.2 Index ................................................................................................................................60

Notes on Instrument Safety
DirectFRAP General Notes Carl Zeiss
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1NOTES ON INSTRUMENT SAFETY
1.1 General Notes
The DirectFRAP imaging system has been designed, produced and tested in compliance with the
standards DIN EN 61010-1 (IEC 61010-1) "Safety Requirements for Electrical Measuring, Control and
Laboratory Instruments", DIN EN 60825-1 (IEC 60825-1) "Safety of Laser Instruments", and DIN EN
61326-1 "Safety Requirements for Electrical Measurement, Control and Laboratory Equipment – EMC
Requirements".
In combination with the Axio Observer, the instrument meets the requirements of the European Directive
98/79/EC (In- Vitro Diagnostics Directive) and carries the marking.
The DirectFRAP imaging system has to be disposed of in compliance with the WEEE Directive 2012/19/EU.
The warning and information symbols explained below will help you recognize at a glance the type and
purpose of the notes included in this operating manual:
CAUTION
This symbol indicates a potential hazard to the user.
CAUTION
Disconnect the instrument from the power supply before opening!
CAUTION Laser radiation
Laser class 3B, max. 500 mW on Laser Bench Tower, Rack or microscope outputs.
Do not expose yourself to the beam. Avoid radiation exposure to the skin!
CAUTION high energy UV radiation!
Do not expose yourself to the beam. Avoid eye or skin exposure to the laser beam!
CAUTION
Keep combustible and highly flammable materials away from the laser beam.
CAUTION
Hot surface!
CAUTION
Dangerous electrical potential!
ATTENTION
This symbol indicates a potential hazard to the instrument or system.
NOTE
This symbol indicates a note containing special instructions.
Always remember: Time invested in familiarizing yourself with the equipment will pay off
many times in your work.

Notes on Instrument Safety
Carl Zeiss General Notes DirectFRAP
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In addition to this manual of the DirectFRAP imaging system, please observe the detailed information
given in the following operating manuals:
−Operating manual for the Axio Observer microscope (B 46-0111)
−ZEN software description (blue edition) (Online version)
−Operating manual for the installation of peripherals (B 40-650)
−Operating manual for the computer system
−Operating manual for the incubation system S1
−Operating manual for the SVB 1 signal distribution box (M60-2-0032)
−Operating manual for Definite Focus (M60-2-0006)
−Operating manual for Laser-TIRF 3 (M60-2-0022)
−Operating manual for Cell Observer SD (M60-2-0021)
−Operating manual for HXP 120 V

Notes on Instrument Safety
DirectFRAP Safety Instructions Carl Zeiss
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1.2 Safety Instructions
1.2.1 Notes on Laser Radiation
The DirectFRAP imaging system is a class 3B laser system and labeled accordingly (total output
power on the microscope: max. 500 mW). If you use laser instruments, adhere to the provisions
concerning health protection and industrial safety. Users should also ensure that they observe
all relevant legal requirements.
If operated correctly, the DirectFRAP imaging system does not create any risk of radiation exposure for
operating personnel. Nevertheless, you should observe the following warnings:
Inform the laser safety officer before you begin with procedures for initial start-
up of the
DirectFRAP imaging system.
•The system is equipped with a safety key switch.
•Always keep the key
for the laser key switch and, where applicable, the keys for any other
laser starter switches in places which are not accessible to persons who are not authorized to
handle the laser.
•Do not expose yourself to the laser beam!
•Do not place any reflecting objects into the beam path!
•Never open any covers on the microscope system while the laser is in operation!
•
Never look into the laser beam, either with or without optical instrument. Nor should you do
so if you just want to view the specimen. Non-
observance involves the risk of eye
damage!
•Cover all currently not used objective nosepiece mounts and camera ports with the dust-
protective caps provided!
•
Take suitable precautions if the laser radiation produces noxious gases, dust and vapors,
secondary radiation or explosive substances on the specimen.
Observe the additional national laser protection requirements!
Keep combustible and highly flammable materials away from the laser beam.

Notes on Instrument Safety
Carl Zeiss Safety Instructions DirectFRAP
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1.2.2 Installation Notes
The DirectFRAP imaging system must be installed and commissioned by technicians trained in
laser safety and authorized by Carl Zeiss. The client is liable to pay the costs of services
rendered by Carl Zeiss Microscopy GmbH.
To install the DirectFRAP imaging
system, ensure a minimum distance of 0.5 m between the
wall and the rear of the system. This distance is necessary for adjustment and maintenance.
Do not set up the equipment near heat sources, such as heaters or direct sunlight. Do not
cover ventilation slots on the microscope system to avoid heat build-up.
The power plug must be inserted in outlets with a grounding (earth) contact. The protective
capacity must not be rendered ineffective by using an extension cable without a ground wire.
Before connecting the power cables, verify that the available line voltage conforms to the
voltage specified on the type label of the laser bench tower or rack.
Always disconnect the power cable before opening the device and replacing fuses.
Only use fuses
which comply with the specifications on the fuse carriers and in this manual.
The use of makeshift fuses and short-circuiting of fuse holders are not permitted.
For reasons of laser safety, the C mount outputs of the microscope must be fitted either with
a camera or closed by a protective cap.
If any protective measures are no longer effective, the device must be taken out of service and
secured against inadvertent operation. Please contact a Carl Zeiss service agency or the Carl
Zeiss Microscopy Service to repair the instrument.
Immersol 518 F® immersion oil irritates the skin. Avoid any contact with your skin, eyes or
clothing.
In case of actual skin contact, wash off with plenty of water and soap. After eye contact,
immediately rinse the eye wi
th plenty of water for at least five minutes. If irritation persists,
consult a physician.
Proper disposal of Immersol 518 F®: Ensure that immersion oil does not enter surface water or
the sewage system.
Do not touch the hot lamp housings and adhere to a cooling time of about 15 minutes.
The device contains electronic components. At the end of its service life, it has to be disposed
of in compliance with the relevant national legislation.

Notes on Instrument Safety
DirectFRAP Safety Instructions Carl Zeiss
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Defective instruments are not to be disposed of as ordinary domestic waste. They should be
disposed of in accordance with the relevant regulations.
Samples should also be disposed of in compliance with the relevant legal requirements and
internal operating procedures.
The DirectFRAP imaging system must be connected to the power supply using several multiple
sockets connected separately to installation sockets. Connection via a single multiple socket
plugged into a single installation socket would lead to inadmissibly high contact current levels.
Maintenance or repair work as well as the modification, removal or replacement of
components not described in this manual, or any other interventions in the equipment may
only be carried out by the manufacturer Carl Zeiss Microscopy GmbH or by persons expressly
authorized by this company. This applies in particular to the microscope system, the laser
manipulation slider, the Laser Bench Tower or Laser Bench Rack, the control PC system and
the cable connections.
Only trained specialists or service staff are permitted to open the instrument.
Operation of the DirectFRAP imaging system is permitted in closed rooms only.
Please consider that the DirectFRAP imaging system is a high-precision opto-electronic
instrument. Improper handling of the instrument may
easily affect its performance or even
damage it and will void all warranty claims.
Microscope tube openings or other ports not used must always be protected against dust and
humidity by mounting the corresponding instrument components or dust caps.
Dirt and dust may impair the performance of the devices. They must therefore be protected
from such influences to the greatest possible extent.
Ensure that the generation of electrostatic charges is avoided by a suitable design of the
workplace environment.
The instruments are not equipped with any special devices for protection against substances
that are corrosive, potentially infectious, toxic or radioactive, or other substances that could be
hazardous to health. All legal regulations must be observed when handling such substances,
particularly the relevant national accident prevention regulations.
Do not operate the supplied equipment in areas in which there is a danger of explosion, in the
presence of volatile anesthetics or flammable solvents, such as alcohol, benzene, etc.
Keep combustible and highly flammable materials away from the laser beam.
Do not replace detachable power cables by power cables with inadequate specifications.
Only the specified power cables should be used.

Notes on Instrument Safety
Carl Zeiss Safety Instructions DirectFRAP
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Portable multiple sockets may not be used. Cables are to be laid such that there is no risk of
tripping, or they are to be covered.
Never disconnect the power cable while the device is in use. Use the power switch to turn the
devices off.
To ensure proper function of the DirectFRAP imaging system, it should be subjected to an
annual safety inspection. The safety checks must be carried out by an authorized Carl Zeiss
service technician. All national safety checks should be carried out.
Health and safety protection regulations should be observed when handling the HXP 120 V
UV radiation source. Relevant national laws should also be complied with.
−
Follow the operating manual for the HXP 120V illumination unit provided by the
manufacturer.
−
Follow the manufacturer's instructions for changing the lamp. Otherwise lamp changes
carry a risk of burns or explosion.
−
Users should not expose themselves to the UV radiation of the HXP 120 V illumination
unit.
−Never look directly into the optical
waveguide when the HXP 120 V illumination unit is
turned on. Non-observance carries the risk of irreversible damage to the eye.
In order to prevent vibrations during operation, the DirectFRAP imaging system should be used
in connection with the Axio Observer anti-vibration plate.
1.2.3 Notes on Computer and Software
Laser manipulation requires that a monitor is attached to the system. That means that you need a PC
with a monitor connected to it.
Additionally, a software program for the digital camera, for image recording and image display on the
monitor must be installed on the control PC.

Notes on Instrument Safety
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1.2.4 Service Interfaces
Fig. 1-1 Service interfaces between Laser Bench Tower and laser manipulation slider
CAUTION
The fiber mounts (Fig. 1-1/1 and 2) may be detached by Carl Zeiss Service staff only.
If one fiber end is not firmly connected to the fiber mount, laser class 3B radiation will emerge!
Non-observance involves the risk of eye damage!

Notes on Instrument Safety
Carl Zeiss Safety Instructions DirectFRAP
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Fig. 1-2 Service interfaces between Laser Bench Rack 3F and laser manipulation slider (DirectFRAP),
Laser TIRF slider (optional) and Cell Observer SD (optional)
CAUTION
The fiber mountings in the Laser Bench Rack (Fig. 1-2/1
), on the laser manipulation slider
(Fig. 1-2/3) and on the optional Laser TIRF slider (Fig. 1-2/2) and Cell Observer SD (Fig. 1-2/4)
units may only be loosened by Carl Zeiss service technicians.
If one fiber end is not firmly connected to the fiber mount, laser class 3B radiation will emerge!
Non-observance involves the risk of eye damage!

Notes on Instrument Safety
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Fig. 1-3 Retaining ring - camera adapter (with protective cover) service interface on the sideport of
the microscope
CAUTION
The screws (Fig. 1-3/1) on the retaining ring (Fig. 1-3/2) for the camera adapter should only be
unscrewed by Carl Zeiss service technicians!
Non-observance involves the risk of eye damage!
If no camera is
mounted on the sideport, it should be sealed by the protective cover
(Fig. 1-3/3), otherwise it is not possible to open the laser shutter.

Notes on Instrument Safety
Carl Zeiss Safety Instructions DirectFRAP
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Fig. 1-4 Camera adapter - camera / protective cover customer interface
The camera adapter (Fig. 1-4/1) is equipped with an interlock connection (two reflected light
barriers) which automatically closes the laser shutter as soon as the camera (Fig. 1-4/2) or the
protective cover (Fig. 1-3/3) is removed from the camera adapter.

Notes on Instrument Safety
DirectFRAP Safety Instructions Carl Zeiss
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Fig. 1-5 Microscope illumination connector - HXP 120 coupling adapter service interface;
Service interfaces for HXP 120 illumination unit;
Sensor ring - HXP 120 coupling adapter / protective cap
A) Microscope illumination connector - HXP 120 coupling adapter service interface
CAUTION
The screws (Fig. 1-5/1) on the sensor ring (Fig. 1-5/2) and the screw (Fig. 1-5/6) on the contact
ring (Fig. 1-5/4) may only be unscrewed by Carl Zeiss service technicians!
Only screw (Fig. 1-5/7) may be unscrewed.
Non-observance involves the risk of eye damage!
B) Service interfaces for HXP 120 illumination unit
1) HXP 120 optical waveguide - HXP 120 coupling adapter
2) HXP 120 optical waveguide - HXP 120 illumination unit
CAUTION
The screws (Fig. 1-5/8) of the safety clamp (Fig. 1-5/9
) for the optical waveguide on the
coupling adapter and the same safety clamp on the illumination unit HXP 120 may only be
unscrewed by Carl Zeiss service technicians!
Non-observance involves the risk of eye damage!
C) Microscope sideport – HXP 120 coupling adapter service interface
The sensor ring is equipped with interlock contacts which automatically close the laser shutter
unless the HXP 120 is connected via the coupling adapter (Fig. 1-5/5
) and the contact ring
(Fig. 1-5/4). If the HXP 120 is not used, the sensor ring (Fig. 1-5/2) must be sealed with the
protective cap (Fig. 1-5/3) to prevent the laser from being used. Only the screw (Fig. 1-5/7) on
the illumination connector of the microscope may be tightened or loosened for assembly /
dismantling of the HXP 120.

Notes on Instrument Safety
Carl Zeiss Safety Instructions DirectFRAP
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Fig. 1-6 Transmitted light carrier - HAL lamp service interface
CAUTION
The clamping screw (Fig. 1-6/1) may be detached by Carl Zeiss Service staff only.
If the lamp housing is removed, laser class 3B radiation will emerge!
Non-observance involves the risk of eye damage!
CAUTION
For changing the halogen lamp of the HAL illuminator, the laser must be switched off in any
case!
Non-observance involves the risk of eye damage!

Notes on Instrument Safety
DirectFRAP Safety Instructions Carl Zeiss
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1.2.5 Warning and Information Labels on the Instrument
Observe the warning and information labels affixed to the DirectFRAP imaging system and Axio Observer
microscope. Check that all these labels are attached. If one of the labels is missing, contact Carl Zeiss
Microscopy GmbH for a free replacement. You will receive replacements free of charge.
Fig. 1-7 Warning and information labels on the DirectFRAP imaging system

Notes on Instrument Safety
Carl Zeiss Safety Instructions DirectFRAP
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Fig. 1-8 Warning and information labels on the laser manipulation slider
Fig. 1-9 Warning and information labels on the laser bench rack

Notes on Instrument Safety
DirectFRAP Safety Instructions Carl Zeiss
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Fig. 1-10 Warning and information labels on the laser bench tower

Notes on Instrument Safety
Carl Zeiss Notes on Warranty DirectFRAP
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Fig. 1-11 Warning and information labels at the SVB 1
1.3 Notes on Warranty
The manufacturer guarantees that the instrument is free of material- or manufacturing-defects when
delivered. Possible defects must be notified to us immediately and steps taken to minimize damage. If
notified of such a defect, the manufacturer is obligated to rectify it at his discretion, either by repairing
the instrument or delivering a fully functioning replacement. No guarantee is provided for defects caused
by natural wear (wearing parts in particular) and improper use.
The instrument manufacturer shall not be liable for damage caused by faulty operation, negligence or
any other tampering with the instrument, particularly the removal or replacement of instrument
components, or the use of accessories from other manufacturers. Such actions will render any warranty
claims invalid.
The customer is not allowed to carry out any maintenance or repair work to the DirectFRAP imaging
system apart from that specified in this operating manual. Repairs are only permitted to the Carl Zeiss
Service or persons especially authorized by it. If any defects occur with the instrument, please get first in
touch with your local Carl Zeiss representative.