Zeiss LSM 510, LSM 510 META Operating Manual
LSM 510 and LSM 510 META
Laser Scanning Microscopes
Operating Manual
Release 3.2
INTRODUCTION LSM 510
Carl Zeiss LSM 510 META
Knowledge of this manual is required for the operation of the instrument. Would you therefore please
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.
Developed in
Collaboration with the European Molecular Biology Laboratory (EMBL)
PF 102209
Meyerhofstr. 1
69012 Heidelberg
GERMANY
Phone: ++49-6221-387-0
Telefax: ++49-6221-387-306
Issued by Carl Zeiss
Advanced Imaging Microscopy
07740 Jena
GERMANY
Phone: ++49-3641 64-34 00
Telefax: ++49-3641 64-31 44
E-mail: micro@zeiss.de
Internet: www.zeiss.de/lsm
Number of this manual: B 45-0008 e
Date of issue: 10/2002
II B 45-0008 e 10/02
LSM 510 INTRODUCTION
LSM 510 META Carl Zeiss
How to make best use of the LSM 510 operating instructions
A few symbols in these operating instructions will help you to recognize the nature and purpose of
information immediately:
The WARNING symbol warns against hazards for the user that might arise when operating the
laser.
This WARNING symbol warns against hazards from dangerously high voltages.
The CAUTION symbol warns against faults and hazards that might arise during operation and
which might cause damage to the unit.
☞
Depending on the problem, these operating instructions will supply you with various possibilities:
• If you want to know where to find certain general areas of information, refer to the following outline
of sections to get a general overview.
• You will find a detailed table of contents at the start of every chapter. There you will see at a glance
what topics are dealt with in detail.
Always remember: The time you invest in getting acquainted with the product will pay
The NOTE symbol will help you to optimally solve your work problem. It represents a practical
tip which will help you to find out which settings and methods are capable of improving or
accelerating a procedure.
The HOT SURFACE symbol warns against hazards for the user that might arise when touching
the lamp housing during operation.
The MAINS PLUG symbol remembers service personal to pull the mains plug before opening
the device housing.
for itself many times over in your application task.
B 45-0008 e 10/02 III
INTRODUCTION LSM 510
Carl Zeiss LSM 510 META
Scope of Equipment Supplied
Country: ..............................................
Order number: ..............................................
Serial number: ..............................................
Delivery date: ..............................................
Custom configuration: ..............................................
Axioskop 2 FS mot 000000-1056-2308
Axioplan 2 imaging
Axiovert 200 M SP 000000-1115-092
Axiovert 200 M BP 000000-1115-094
Axiovert 200 M BP/SP 000000-1115-289
Objectives: ..............................................
..............................................
..............................................
Confocal Laser Scanning Module LSM 510
Configuration 1 000000-1219-975
Configuration 2 000000-1219-977
Configuration 3 000000-1219-978
Configuration 4 000000-1219-979
Configuration 5 000000-1219-980
Configuration 9 000000-1219-981
Configuration 11 000000-1219-982
Configuration 12 000000-1219-983
Configuration 13 000000-1219-984
Configuration 15 000000-1219-985
Configuration 16 000000-1219-986
Configuration 18 000000-1219-987
Configuration 3 META 000000-1219-995
Configuration 13 META 000000-1219-997
Configuration 15 META 000000-1220-000
Configuration 18 META 000000-1220-004
Control computer 000000-0438-360
21“ monitors 000000-1060-089
TFT monitors 000000-0435-035
MOT (ie) 000000-1166-265
The license to the LSM control software is included in each configuration 1...18 (META).
IV B 45-0008 e 10/02
LSM 510 INTRODUCTION
LSM 510 META Carl Zeiss
Optional software:
Image VisArt option 000000-1207-934
3D for LSM option 000000-1207-919
3D Deconvolution option 000000-1207-920
Physiology option 000000-1207-930
Topography option 000000-1207-933
Multiple Time Series option 000000-1207-926
Macro Reader and Editor option 000000-1207-925
StitchArt option 000000-1207-932
Canon S 830 D Photo printer 000000-0445-508
Kodak XLS 8650 PS printer 000000-1113-131
Transparency exposure device (D) 412410-9011-000
Large system table 453031-0000-000
Small system table 453032-0000-000
System baseplate 000000-1171-342
Active anti-vibration table 30" x 30" 000000-1177-841
Active anti-vibration table 1200 mm x 1400 mm 000000-1198-394
Active anti-vibration table 1800 mm x 1400 mm 000000-1150-353
UV laser 412410-9015-000
XY scanning stage for Axiovert 200 M BP 000000-1113-509
XY scanning stage for Axioplan 2 imaging
MOT 000000-1027-823
Piezo objective drive W = 0.8" 000000-1210-045
High resolution Z stage HRZ 200 for Axiovert 200 M 000000-1115-248
High resolution Z stage HRZ 200 for Axioplan 2 imaging
MOT (ie) 000000-1115-245
Set of INDO filters 447960-0000-000
Set of SNARF filters 447961-0000-000
AxioCam HRm 000000-0445-553
AxioCam HRc 000000-0412-312
B 45-0008 e 10/02 V
INTRODUCTION LSM 510
Carl Zeiss LSM 510 META
The LSM 510 in the configuration as checked above
was installed and handed to the customer in functional condition
on ................................................
by ................................................
Phone: ................................................
Fax: ................................................
The customer has been instructed on how to operate and
maintain the equipment.
(Place)......................................................, (date) .....................................
...............................................................
Carl Zeiss Jena GmbH
Microscopy Division
...............................................................
Customer
One copy to be kept by customer
One copy to be kept by Carl Zeiss
VI B 45-0008 e 10/02
LSM 510 INTRODUCTION
LSM 510 META Carl Zeiss
1
2
3
4
5
Notes on Device Safety
This section contains general notes on device safety, safe operation, and possible hazards
caused by failure to observe the instructions.
LSM 510 - Setup Requirements
The Setup Requirements section outlines the installation and supply requirements of the
LSM 510 Microscope System, together with the relevant specifications.
Introduction to Laser Scanning Microscopy
Here you will find an introduction to Laser Scanning Microscopy, with an explanation of the
principles of confocal imaging. The section also outlines the ways to present LSM image series
in three dimensions, and introduces you to the performance features of your LSM 510.
Quickstart
Operation in Expert Mode
In the Operation section you will find the most important steps and procedures of the
LSM menu structure. The step-by-step description how to get an image will be shown by
typical application examples including the WINDOWS NT 4.0 graphic user environment.
6
7
8
9
10
11
VBA Programming for LSM
Routine Mode and Tools
This section contains a description of the Routine Mode for scanning images using the
LSM scanning module and the use of the tools for setting the microscope.
3D for LSM 510
This section contains a description of the LSM 3D software package (basic program and addons). At the same time, all functions and settings are presented in a systematic form and in
the order in which they can be reached from the basic menu via sub-menus and dialog boxes.
Annex
The annex contains the Application-specific Configurations and special notes and information
for using the LSM microscope.
Multiphoton Laser Scanning Microscopy - Using the Zeiss LSM 510 NLO
Certification
B 45-0008 e 10/02 VII
INTRODUCTION LSM 510
Carl Zeiss LSM 510 META
VIII B 45-0008 e 10/02
LSM 510 NOTES ON DEVICE SAFETY
LSM 510 META Contents Carl Zeiss
CHAPTER 1 NOTES ON DEVICE SAFETY
CONTENTS
Page
1 NOTES ON DEVICE SAFETY...................................................................................................1-3
1.1 General...................................................................................................................................1-3
1.2 Regulations .............................................................................................................................1-3
1.3 Notes on Setting up the Microscope System.............................................................................1-4
1.4 Notes on Handling the Computer and Data Media................................................................... 1-5
1.5 Notes on Care, Maintenance and Service .................................................................................1-6
1.6 Notes on Handling the Laser Components ............................................................................... 1-7
1.7 Warning and Information Labels..............................................................................................1-8
B 45-0008 e 10/02 1-1
NOTES ON DEVICE SAFETY LSM 510
Carl Zeiss Contents LSM 510 META
1-2 B 45-0008 e 10/02
LSM 510 NOTES ON DEVICE SAFETY
LSM 510 META General / Regulations Carl Zeiss
1 NOTES ON DEVICE SAFETY
1.1 General
The LSM 510 laser scanning microscope, including its original accessories and compatible accessories
from other manufacturers, may only be used for the purposes and microscopy techniques described in
this manual (intended use).
The manufacturer will not assume liability for any malfunction or damage caused by any thing
other than the intended use of the LSM 510 or individual modules or parts of it, nor by any
repair or other service operation performed or attempted by persons other than duly
authorized service staff. Any such action will invalidate any claim under warranty, including
parts not directly affected by such action. This also includes the modification of the system
computer with new cards, etc. by the user.
1.2 Regulations
Extensive knowledge of the hardware/the system is indispensable for safe operation of the LSM 510.
☞
⇒
The LSM 510 has been designed, built and tested in conformity with the standards DIN EN 61010-1
⇒
As the system is largely operated via menus on a computer, you should be acquainted with the
⇒
In accordance with WHO regulations, the LSM 510 is a device that belongs to laser hazard class 3 B.
Read these operating instructions and all device publications belonging to the system
conscientiously before operating the LSM 510! You can obtain additional information on the
hardware configuration delivered and on optional system extensions from the manufacturer or
via the service hotline.
(IEC 1010-1) "Safety requirements for electrical instrumentation and control and laboratory
apparatus", and DIN EN 60825-1 (IEC publication 825-1) "Safety of laser equipment", and taking
relevant CSA and UL specifications into account.
principles of the operating system and its WINDOWS NT 4.0 and WINDOWS 2000 graphical user
interface. The respective manuals are supplied together with the programs.
WHO recommendations concerning health and industrial protection when handling laser devices
must be observed. The operator of the unit must also observe all and any relevant statutory
accident prevention regulations.
B 45-0008 e 10/02 1-3
NOTES ON DEVICE SAFETY LSM 510
Carl Zeiss Notes on Setting up the Microscope System LSM 510 META
1.3 Notes on Setting up the Microscope System
☞
The LSM 510 laser scanning microscope is delivered in several crates:
• Crate 1: microscope stand, laser scanning module, control unit
• Crate 2: computer
• Crate 3: monitor
• Crate 4: large system table
• Crate 5: second microscope stand
• Crate 6: small system table
• Crate 7: scan module META
• Crate 8: upgrade kit META
Do not set up the unit in the proximity of heat sources such as radiators or direct sunlight. To avoid heat
build-ups, the ventilation louvers on the microscope system must not be covered up.
The unit must be connected to a properly installed socket outlet with earthing contact by means of the
mains cables supplied. Continuity of PE connection must not be affected by the use of extension leads.
Setting up, assembly on the system base plate and commissioning of the LSM 510 must be
performed by authorized Carl Zeiss service staff, who are also advised to give the customer's
operators a basic introduction to operation and maintenance.
The LSM 510 must be set up so as to ensure that the minimum clearance between the wall
and the rear of the system is no less than 0.5 m. This clearance is needed for adjustment and
maintenance operations.
Before connecting the mains cables, please check whether your mains voltage corresponds to
the voltage specified on the rating plate of the laser module.
For reasons of laser safety, the TV port on the microscope must either be equipped with a
camera or covered by a cap.
Maintenance, repairs, modifications, removal or exchange of components, or other
interference with the equipment beyond the operations described in this manual may only be
carried out by the manufacturer Carl Zeiss or by persons expressly authorized by us to do so.
This applies especially to the microscope system, the laser scanning module, lasers, the PC
system, the power supply units, cable connections and other system components.
Please note that the LSM 510 is a high-precision opto-electronic instrument. Inexpert handling
may easily impair it’s function or even damage it.
After installation or after conversion of the LSM system, authorized specialized staff must carefully check
that it is in a proper condition and, particularly, that covers protecting against laser radiation are
provided.
Tube openings or other unused mounts should always be protected against dust and moisture with the
corresponding device components or with termination covers/blind plugs.
By establishing a corresponding workplace environment, please ensure that the formation of electrostatic
charges by electronic components is avoided.
To avoid vibrations during operation, the LSM 510 should only be operated in conjunction with the
system table (vibration damping).
1-4 B 45-0008 e 10/02
LSM 510 NOTES ON DEVICE SAFETY
LSM 510 META Notes on Handling the Computer and Data Media Carl Zeiss
1.4 Notes on Handling the Computer and Data Media
The computer used as standard in your LSM system is an IBM-compatible high-end pentium computer
with WINDOWS NT 4.0 or WINDOWS 2000 operating system.
As standard, your computer has one hard disk drive, one drive for 1.44 MB diskettes and one CD-ROM
drive. An CD reader/writer is installed.
☞
☞
Do make sure, though, that you receive your LSM system with the operating system installed,
with initialization and start files set up and with the LSM program also installed.
When working with the hard disk, it is important to know that the more data it contains, the
slower its operation will become. Therefore, data that you do not need permanently should be
stored on a diskette or CD-ROM.
When handling diskettes, avoid data losses by protecting them against extreme temperatures,
moisture and magnetic fields. The data on a diskette is stored in the form of magnetic signals.
To some extent, monitors, telephones or even lamps generate magnetic fields that might
destroy this data. Also, never open the metal cover on diskette cases. A diskette´s surface can
also be destroyed by touching it.
Never turn your computer off before you have exited the LSM program and run down the
WINDOWS NT operating system. Otherwise, the program and/or data files may get lost.
When handling discs of the CD reader/writer, do not touch the data side of the disc (the side
of the disc with no label or printing).
Do not apply paper labels or write on any part of the disc, data side or label side. If dust or
fingerprints get on the disc, wipe it with a soft cloth from the center to the edge, but do not
use benzine, paint thinner, record cleaner, or static repellent. This can damage the disc.
Do not place the disc in any place where it is exposed to direct sunlight or high temperatures.
B 45-0008 e 10/02 1-5
NOTES ON DEVICE SAFETY LSM 510
Carl Zeiss Notes on Care, Maintenance and Service LSM 510 META
1.5 Notes on Care, Maintenance and Service
The manufacturer of the unit cannot be held liable for damage resulting from operating errors,
negligence or unauthorized tampering with the device system, particularly as the result of removal or
replacement of parts of the unit or as the result of the use of unsuitable accessories from other
manufacturers.
Any such action will also render all warranty claims null and void.
You are well advised to arrange a service agreement with your nearest Zeiss representative to guarantee
perfect functioning of the microscope system in the long term.
Modifications and conversion work on the components of the system must only be carried out by the
manufacturer, by the service agency or by persons authorized and trained for this purpose by the
manufacturer.
Damaged units or parts may only be repaired or maintained by the responsible service agency.
Care operations that may be carried out by operating staff are limited to cleaning painted and glass
surfaces.
• Cleaning painted surfaces
To do this, use a clean cloth that has been moistened in a mixture of water and some detergent; do
not use any solvent, however. Dry with a lint-free cloth.
• Cleaning glass surfaces
Glass surfaces that have become soiled or which are marked with fingerprints may be rubbed with a
clean optical cleaning cloth.
If soiling is persistent, dip the optical cleaning cloth into a mixture of distilled water and a small
quantity of detergent.
To complete cleaning, lightly breathe on the glass surface and rub it dry with a clean cloth. Lint or
dust is best removed with a clean hairbrush.
The air filter mat at the bottom of the LSM 510 Control Unit must be cleaned every six months.
Filter mats can be ordered from our Service Department.
1-6 B 45-0008 e 10/02
LSM 510 NOTES ON DEVICE SAFETY
LSM 510 META Notes on Handling the Laser Components Carl Zeiss
1.6 Notes on Handling the Laser Components
The LSM 510 is a laser hazard class 3 B instrument and is marked as such.
This moderate-risk class embraces medium-power lasers. You must take care not to expose
yourself to the radiation of such lasers. In particular, never look into the laser beam!
The following laser types are currently recommended for use in the LSM 510.
1 Ar 351/364 (UV)
2 Ar/ML 458/477/488/514
3 HeNe 543
4 ArKr 488/568
5 HeNe 633
☞
If used properly, the LSM 510 will not pose any laser radiation risks for operating staff. The dangerous
laser radiation area is limited to the beam path and to a distance of up to around 10 cm from the
specimen. Nevertheless, you should observe the following warnings:
Please contact Carl Zeiss if you intend to use a laser type with a wavelength other than the
ones above.
• If necessary - insofar as specified by law - inform the laser protection officer before
commissioning the laser.
• Always store laser key switches and, if applicable, keys for further laser power supply units,
where they are inaccessible to persons not authorized to operate the laser.
• Do not place any reflecting objects into the beam path.
• Never open any covers or panelings.
• Never look into the laser beam, not even to simply view the specimen, whether with the aid
of optical instruments or without. Otherwise you risk going blind!
• Do not leave any screw positions of the nosepiece uncovered.
☞
B 45-0008 e 10/02 1-7
Suitable protective measures must be taken if gases, dust or vapors hazardous to health,
secondary radiation or explosive objects should arise on the specimen as a result of laser
radiation.
NOTES ON DEVICE SAFETY LSM 510
Carl Zeiss Warning and Information Labels LSM 510 META
1.7 Warning and Information Labels
The warning and information labels attached on the LSM 510 must be observed. Check
whether all of the labels shown below are provided on your instrument, and contact Carl Zeiss
Germany or one of the service agencies if you should discover that any of the labels should be
missing. You will receive a free replacement.
The
label means: "Do not remove securing screw as otherwise laser beam will escape. For
use by service only!"
Fig. 1-1 Warning and information labels on the Axiovert 200 M microscope with the LSM 510
scanning module
1-8 B 45-0008 e 10/02
LSM 510 META NOTES ON DEVICE SAFETY
LSM 510 Warning and Information Labels Carl Zeiss
Fig. 1-2 Warning and information labels on the Axioplan 2 imaging MOT microscope with LSM 510
META scanning module
B 45-0008 e 10/02 1-9
NOTES ON DEVICE SAFETY LSM 510
Carl Zeiss Warning and Information Labels LSM 510 META
Fig. 1-3 Warning and information labels on laser components (page 1)
1-10 B 45-0008 e 10/02
LSM 510 META NOTES ON DEVICE SAFETY
LSM 510 Warning and Information Labels Carl Zeiss
C
O
H
E
R
E
n
T
E
n
T
E
R
P
R
IS
E
PEAK
maximum
water
pressure
SEARCH
60 p.s.I.
E4
OUT
E
n
T
E
R
P
R
IS
E
C
O
H
E
R
E
n
T
water
J37
E5
E4
J6
Fig. 1-3 Warning and information labels on laser components (page 2)
B 45-0008 e 10/02 1-11
NOTES ON DEVICE SAFETY LSM 510
Carl Zeiss LSM 510 META
1-12 B 45-0008 e 10/02
LSM 510 LSM 510 - SETUP REQUIREMENTS
LSM 510 META Contents Carl Zeiss
CHAPTER 2 LSM 510 - SETUP REQUIREMENTS
CONTENTS
Page
2 LSM 510 - SETUP REQUIREMENTS................................................................................ 2-3
2.1 Space Requirements ........................................................................................................2-3
2.1.1 LSM (one microscope, large system table): 300 × 250 cm ................................................. 2-3
2.1.2 LSM with Ar UV Laser......................................................................................................2-4
2.1.3 LSM prepared for Two Photon Lasers (NLO)......................................................................2-6
2.2 Power Requirements........................................................................................................2-8
2.2.1 Phase 1 (LSM).................................................................................................................. 2-9
2.2.2 Phase 2 (LSM, Power 3) ...................................................................................................2-9
2.2.3 Separate Connection .......................................................................................................2-9
2.3 Physical Dimensions....................................................................................................... 2-10
2.4 Dimension of Shipment Crates....................................................................................... 2-11
2.5 Environmental Requirements..........................................................................................2-11
2.6 Vibrations...................................................................................................................... 2-12
2.7 Laser Specifications........................................................................................................ 2-12
2.7.1 Coherent Enterprise 653 II: 352, 364 nm, 80 mW, Laser Class 3 B ..................................2-12
2.7.2 Point Source i-flex 2000: 405 nm, 25 mW, Laser Class 3 B..............................................2-12
2.7.3 LASOS LGK 7786 P / Power supply 7460 A: 543 nm, 1 mW, Laser Class 3 B ...................2-13
2.7.4 LASOS LGK 7628-1: 633 nm, 5 mW, Laser Class 3 B ...................................................... 2-13
2.7.5 LASOS LGK 7812 ML-4 / LGN 7812: 458, 477, 488, 514 nm, 30 mW, Laser Class 3 B.....2-13
2.7.6 Melles Griot 643-YB-A02 / Power supply 171B: 488, 568 nm, 30 mW, Laser Class 3 B.... 2-13
2.7.7 AOTF.............................................................................................................................2-13
2.8 Microscopes ..................................................................................................................2-14
2.9 Scanning Module........................................................................................................... 2-15
2.10 Laser Module VIS (405, 458, 477, 488, 514, 543, 633 nm).............................................2-16
2.11 Laser Module UV (351, 364 nm) ....................................................................................2-16
2.12 System Overview LSM 510 META...................................................................................2-17
2.13 System Overview LSM 510 META - NLO .........................................................................2-19
B 45-0008 e 10/02 2-1
LSM 510 - SETUP REQUIREMENTS LSM 510
Carl Zeiss Contents LSM 510 META
2-2 B 45-0008 e 10/02
LSM 510 LSM 510 - SETUP REQUIREMENTS
LSM 510 META Space Requirements Carl Zeiss
2 LSM 510 - SETUP REQUIREMENTS
2.1 Space Requirements
2.1.1 LSM (one microscope, large system table): 300 × 250 cm
Power Plug
System rack
1000
Large system table
1500
3000
Fig. 2-1
500
2500
800
The system rack contains the laser module (Helium-Neon laser 543 nm, 633 nm and Argon ion laser), the
power supply for the Argon ion laser, for HBO lamp and halogen lamp, the electronic control unit (ECU)
and the MCU28 unit (if a motorized XY stage is applied).
B 45-0008 e 10/02 2-3
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