LEICA TCS SP5 User Manual

Leica TCS SP5
User Manual
Leica Microsystems CMS GmbH Am Friedensplatz 3 68165 Mannheim Germany
1 Notes about this manual page 1.............
1.1 Copyright page 1..............................................
2 General page 3...........................
2.1 About these operating instructions page 3.........................
3 Proper intended use page 5.................
4 Legal Notes page 7........................
4.1 DISCLOSURE page 7..........................................
4.2 REVISIONS AND CHANGES page 8.............................
4.3 WARRANTY page 8............................................
4.4 Patents page 8................................................
4.5 TRADEMARKS page 10.........................................
Table of Contents
4.6 Software licenses page 10.......................................
5 TCS SP5 specifications page 11..............
5.1 System overview page 11........................................
5.2 Dimensions page 12.............................................
5.2.1 System with inverted stand: page 12...............................
5.2.2 System with upright stand page 13................................
5.3 Installation site requirements page 13..............................
5.4 Permissible ambient conditions page 14............................
5.5 Electrical connections page 14....................................
5.6 Required cooling capacity page 16................................
5.7 Important additional notes page 16................................
5.8 Required safety measures for VIS and UV systems page 17..........
5.9 Required laser safety measures for MP systems page 17............
5.10 Important additional notes page 17................................
6 Safety Notes page 19.......................
6.1 Which standards does this product meet? page 19..................
6.2 Which laser class does the product have? page 21..................
6.3 Laser class for VIS and UV systems page 21.......................
6.4 Laser class for MP systems page 21..............................
6.5 Warnings, Safety Cautions, and Notes page 22.....................
6.6 What should the user of the laser scanning microscope observe? page 23
6.7 Safety Notes for the User page 25................................
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Table of Contents
6.8 Safety Notes for Operation page 26...............................
6.9 Specific Safety Notes page 30....................................
6.9.1 Operational reliability page 30....................................
6.9.2 De-energizing the system page 30................................
6.9.3 Maximum current load of the power outlet strip at the supply unit page 31
6.10 Laser safety devices page 32.....................................
6.10.1 Eye protection for VIS or UV systems page 32......................
6.10.2 Detachable-key switch page 33...................................
6.10.3 Emissions warning indicators page 34.............................
6.10.4 Remote interlock connector on the supply unit page 36...............
6.10.5 Remote interlock connector on the external 405nm advanced laser page 37
6.10.6 Remote interlock connectors on additional external lasers page 37....
6.10.7 Interlock connection on scanner page 39...........................
6.10.8 Function and position of safety switches page 40....................
6.10.9 Transmitted-light lamp housing for inverted stands page 40...........
6.10.10 Transmitted-light lamp housing for upright stands page 42............
6.10.11 Mirror housing on upright stand page 43...........................
6.10.12 Special laser safety equipment page 44............................
6.10.12.1 Safety beam guide and beam collector page 44.....................
6.10.12.2 Eye protection for MP systems page 45............................
6.10.12.3 Shielding in MP systems (UV laser) page 46.......................
6.11 Overview of usable VIS/UV lasers page 47.........................
6.12 Overview of usable MP lasers (IR) page 48.........................
6.13 Safety label on TCS SP5 system page 49..........................
6.13.1 Inverted stand DMI 4000/6000 CS page 49.........................
6.13.2 Upright stand DM 5000/6000 CS page 50..........................
6.13.3 Scan head page 53.............................................
6.13.4 Supply unit page 54.............................................
6.13.5 External 405-nm laser Advance / 405-nm imaging laser page 55......
6.13.6 External 488-nm laser page 56...................................
6.13.7 MP beam coupling unit page 57...................................
6.13.8 Cover (for replacement flange) page 58............................
6.14 Requirements related to the installation/storage location page 59......
6.15 Changing the installation site page 60..............................
6.16 Scanner cooling page 61.........................................
7 Care and cleaning page 63..................
7.1 Cleaning the optical system of the microscope page 63..............
7.2 Cleaning the microscope surface page 64..........................
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8 Startup of the system page 67................
8.1 Setting Up Users page 70........................................
9 Starting the LAS AF page 75................
10 Introduction to LAS AF - Help page 79........
10.1 General page 79................................................
10.1.1 Calling Online Help page 79......................................
10.1.2 Structure of the online help page 80...............................
10.2 Imprint page 81.................................................
10.2.1 In the respective context (context-sensitive) page 82.................
10.2.2 Via the Help menu page 82.......................................
10.2.3 Full-text search with logically connected search terms page 83........
10.3 Key combinations page 84.......................................
Table of Contents
11 Structure of the user interface page 85........
1 1.1 General structure of the user interface page 85.....................
12 What is the Köhler illumination? page 87......
12.1 Setting the Köhler Illumination page 88............................
13 Introduction to confocal work page 91........
13.1 Preparation page 91.............................................
13.1.1 The objective page 92...........................................
13.1.2 Conventional microscopy page 93.................................
13.1.3 Why scan? page 96.............................................
13.1.4 How is an optical section created? page 98.........................
13.2 Acquiring optical sections page 100................................
13.2.1 Data acquisition page 101.........................................
13.2.2 Illumination p age 103.............................................
13.2.3 Beam splitting page 104..........................................
13.2.4 Emission bands page 104.........................................
13.2.5 The pinhole and its effects page 106................................
13.2.6 Image detail and raster settings page 108...........................
13.2.7 Signal and noise page 113........................................
13.2.8 Profile cuts page 116.............................................
13.3 Multiparameter fluorescence page 117..............................
13.3.1 Illumination p age 1 17.............................................
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Table of Contents
13.3.2 Beam splitting page 118..........................................
13.3.3 Emission bands page 119.........................................
13.3.4 Crosstalk page 119...............................................
13.3.5 Sequential capture page 121......................................
13.3.6 Unmixing page 121...............................................
13.4 3D Series page 124..............................................
13.4.1 z-stack page 124.................................................
13.4.2 About section thickness... page 124................................
13.4.3 ...and spacing page 125...........................................
13.4.4 Data volumes page 125...........................................
13.4.5 Depictions page 127..............................................
13.4.5.1 Gallery page 127.................................................
13.4.5.2 Movie page 128..................................................
13.4.5.3 Orthogonal projections page 128...................................
13.4.5.4 Rotated projections page 129......................................
13.5 Time series page 131.............................................
13.5.1 About scan speeds page 131......................................
13.5.2 Points page 132.................................................
13.5.3 Lines page 132..................................................
13.5.4 Planes page 132.................................................
13.5.5 Spaces (time-space) page 133.....................................
13.5.6 FRAP measurements page 133....................................
13.6 Spectral series page 134..........................................
13.6.1 Data acquisition and utilization page 134............................
13.6.2 About spectral resolution page 134.................................
13.7 Combinatorial analysis page 135...................................
5 Switching off the system page 137.............
6 Contact page 139...........................
7 Declaration of conformity page 141............
8 Glossary page 143..........................
18 Safety data sheets page 153...................
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1 Notes about this manual
1.1 Copyright
The contents of this manual are current as of thetimeofpublicationandaresubjectto change without notice.
Leica Microsystems CMS GmbH assumes no warranty with respect to this manual, inc­luding but not limited to implicit warranties for salability and for suitability for a specific pur­pose. Leica Microsystems CMS GmbH is not liable for errors or accidental damages or in­cidental damages in conjunction with equip­ment, performance or usage of this manual or the examples contained therein.
Notes about this Manual
© Leica Microsystems CMS GmbH
This document contains information that is protected by copyright law. All rights reser­ved. Duplication, adaptation or translation of this manual requires the prior written approval of Leica Microsystems CMS GmbH unless otherwise permitted through copyright arran­gements.
The programs for controlling this product are protected by copyright law. All rights reser­ved. Reproduction, adaptation or translation of these programs w ithout prior written ap­proval of Leica Microsystems CMS GmbH is strictly prohibited.
MicrosoftWindows,WindowsXP,theWin­dows logo and the Windows CE logo are ei-
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Notes about this Manual
ther registered trademarks or trademarks of Microsoft Corporation in the US and/or other countries. Microsoft products are licensed for other companies by Microsoft Licensing, Inc., a subsidiary company that is wholly owned by Microsoft Corporation.
All other tradenames and product names in this document are brands, service brands, trademarks or registered trademarks of their respective owners.
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2 General
2.1 About these operating instructions
The main focus of these operating instruc­tions is directed to the safety instructions that must be observed while working with the la­ser scanning microscope.
In addition, these operating instructions pro­vide a rough overview of the operating princi­ple of laser scanning microscopes. It pre­sents you with the first steps for starting and startup of the system and provides a descrip­tion of the Leica Application Suite Advanced
Fluorescence (LAS AF).
General
The Leica TCS SP5 is supplied with the la­test version of the licensed Leica Application Suite Advanced Fluorescence. To maintain information on the most current level, the description of software functions was inten ­tionally omitted from these operating instruc­tions. Instead, you are referred to the online help of the Leica Application Suite Advanced Fluorescence in which you can obtain the most up-to-date explanations and instruc­tions about the corresponding software func­tions.
Please read the chapter ”Introduction to the help of the Leica Application Suite Advanced Fluorescence” in these operating instructions to familiarize yourself first with setup and operation. Additional information about spe­cific functions can subs equently be obtained electronically at your TCS workstation.
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General
Page 4
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3 Proper intended use
The system was designed for confocal recor­ding (laser scanning images) of fluore­scence-marked living and fixed specimens as well as for quantitative measurements in the area of life science.
This system is intended for use in a lab. Applications of in-vitro diagnostics according
to the medical products law are excluded from proper intended use.
The manufacturer assumes no liability for any improper intended use and for use out­side the specifications of Leica Microsystems CMSGmbHaswellasanyrisksresulting from it. In such cases, the declaration of con­formity becomes invalid.
Proper Intended Use
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Proper Intended Use
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4LegalNotes
Legal Notes
Manufactured in the Federal Republic of Germany.
© Copyright 2005, Leica Microsystems CMS GmbH. All rights reserved.
No part of this publication may be reprodu­ced or transmitted in any form or by any means, electronic or mechanical, including photocopying, recording, or storing in a re­trieval system, or translating into any lan­guage in any form without the express writ­ten permission of Leica Microsystems CMS GmbH. This also includes photocopying, re­cording or storing on a retrievable system as well as the translation into another language.
4.1 DISCLOSURE
This document contains Leica Microsystems CMS GmbH proprietary data and is provided solely to its customers for their express be­nefit of safe, efficient operation and mainte­nance of the product described herein. Use or disclosure of Leica Microsystems CMS GmbH proprietary data for the purpose of manufacture or reproduction of the item described herein, or any similar item, is pro­hibited, and delivery of this document shall not constitute any license or implied authori­zation to do so.
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Legal Notes
4.2 REVISIONS AND CHANGES
Leica Microsystems CMS GmbH reserves the right to revise this document and/or to further develop and improve the products described in this document at any time wi­thout prior notice or any other obligation. Information and specifications in this manual are subject to change without notice.
4.3 WARRANTY
Leica Microsystems CMS GmbH provides this publication ”as is” without warranty of any kind, either expressed or implied, inclu­ding but not limited to the implied warranties of merchantability or fitness for a particular purpose. All reasonable precautions have been taken in the preparation of this document, including both technical and non-technical proofing. Leica Microsystems CMS GmbH accepts no responsibility for any errors or omissions. In addition, Leica Microsystems CMS GmbH is not responsible for direct, incidental or con­sequential damages that could result from the use of the material described in this do­cument.
4.4 Patents
Page 8
The product is protected by the following US patents: 5,886,784; 5,903,688; 6,137,627; 6,222,961;
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Legal Notes
6,285,019; 6,311,574; 6,355,919; 6,423,960; 6,433,814; 6,444,971; 6,466,381; 6,510,001; 6,614,526; 6,654,165; 6,657,187; 6,678,443; 6,687,035; 6,738,190; 6,754,003; 6,801,359; 6,831,780; 6,850,358; 6,867,899. Further patents are pending.
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Legal Notes
4.5 TRADEMARKS
4.6 Software licenses
Throughout this manual, trademarked names may be used. Rather than including a trade­mark (™) symbol at every occurrence of a trademarked name, we state that we are using the names only in an editorial fashion, and to the benefit of the trademark owner, with no intention of infringement.
The software described in this document is furnished under a License Agreement which is included with the product. This agreement specifies the permitted and prohibited uses of the product.
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5 TCS SP5 specifications
5.1 System overview
12
Specifications
6
figure1. System components (overview)
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5
1 TCS SP5 Scanner 2 Control panel 3 TCS workstation 4 Supply unit 5 Panel box 6 Stand with scanner
4
3
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Specifications
5.2 Dimensions
5.2.1 System with inverted stand:
figure2. Dimensions of the TCS SP5 with inverted stand
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5.2.2 System with upright stand
Specifications
figure3. Dimensions of the TCS SP5 with upright stand
5.3 Installation site requirements
Do not expose the system to draft.
Ensure that the system is not installed next to elevators, air conditioners or ventilation systems. For this reason, the installation site should be carefully planned.
Ensure that the e nvironment is as dust­free as possible.
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Specifications
Please also read the notes on protection against dust in the chapters on maintenance and cleaning. Installing the system in darke­ned rooms is also advisable.
For installation, maintenance and trans­port, the TCS SP5 system requires doors with inside spans of 1.00 m.
With regard to the load-bearing capacity of the floor, please note that the system will ap­ply a static load of 200 kg/m@.
5.4 Permissible ambient conditions
Permissible temperature range for operation: 18 ... 25° C
Permissible relative humidity: 20-80% (non­condensing)
Permissible vibration speeds: Frequency range [5 Hz–30 Hz]: less than 30
μm/s (effective value) Frequency range [> 30 Hz]: less than 60 μm/s (effective value)
Pollution degree: Class 2
5.5 Electrical connections
Supply voltage (Input 1/Input 2): 100...240 V AC +/- 10%
Frequency: 50/60 Hz
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Power consumption: 2 x 1.6kW Overvoltage category: II The building installation must feature two se-
parate power connections with the following fuse protection:
D 100 V - 120 V AC power: 20 A
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Specifications
D 200 V - 240 V AC power: 12-16 A For the specifications of external lasers such
as UV and MP lasers, please refer to the ma­nufacturer’s documentation.
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Specifications
5.6 Required cooling capacity
The TCS SP5 system has a maximum power consumption of 3.2 kW (VIS system).
For the specifications of external lasers such as UV and MP lasers, please refer to the ma­nufacturer’s documentation.
5.7 Important additional notes
The optimal optical performance of the sy­stem can be achieved using standard objec­tives and standard immersions only at 22° Celsius + 1 Kelvin.
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Specifications
5.8 Required safety measures for VIS and UV sy­stems
Please observe valid country-specific regula­tions pertaining to laser safety measures for laser class IIIb. The operator is responsible for observing the laser safety regulations.
5.9 Required laser safety measures for MP sy­stems
Please observe valid country-specific regula­tions pertaining to laser safety measures for laser class 4/IV. The operator is responsible for observing the laser safety regulations.
5.10 Important additional notes
The optimal optical performance of the sy­stemcanbeachievedusing standard objec­tives and standard immersions only at 22° Celsius + 1 °Celsius.
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Specifications
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6 Safety Notes
Safety Notes
The instrument is Class 3B or 4 laser equipment (depending on the laser used) in accor­dance with IEC/EN 60225-1. This laser equipment may be operated only by persons who have been trained in the use of the systems and the potential dangers of laser radiation.
As it is not possible to anticipate every po­tential hazard, please take care and apply common sense to the installation, operation and maintenance of this product. Observe all safety precautions relevant to Class IIIb la­sers and Class IV MP systems.
Do not deviate from the operating and main­tenance instructions provided herein. The failure to observe these instructions shall be exclusively at the user’s own risk and may void the warranty.
6.1 Which standards does this product meet?
This device was tested and meets the requi­rements of the following standards:
D IEC/EN 61010-1
”Safety requirements for electrical equip­ment for measurement, control and labo­ratory use”
D IEC/EN 60825-1
”Safety of laser products, Part 1: Equipment classification, require­ments and user’s guide”
D IEC/EN 61326
”Electrical equipment for measurement, control and laboratory use - EMC requi­rements” (Class A). This is a Class A device. Operating this device on a public low-voltage grid may result in radio interference. The operator
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Safety Notes
must take suitable measures should this occur.
D IEC/EN 61000-3-2 (EMC)
”Electromagnetic Compatibility” Part 3-2: Limits - Limits for harmonic currents
D IEC/EN 61000-3-3 (EMC)
”Electromagnetic Compatibility” Part 3-3: Limits - Limits - Section 3: Limits for voltage fluctuations and flik­ker in low-voltage networks.
D For use in the USA:
CDRH 21 CFR 1040.10: Laser Products U.S. Food and Drug Administration (FDA) (”Complies with FDA performance standards for laser products except for deviations pursuant to laser notice No. 50, dated 26 July,
2001.”)
For the scope of the CDRH/FDA (USA), the designation Laser Class 3B in the text must be replaced by IIIb andClass4byIV.
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Safety Notes
6.2 Which laser class does the product have?
Laser
type
VIS 400 - 700 nm,
UV 350 - 700 nm,
MP 350 - 1050 nm,
Wavelength range Configuration Laser class
Combination of lasers
(visible laser
radiation)
(visible and invisible la-
ser radiation)
(visible and invisible la-
ser radiation)
from Chapter 6.11
(without lasers having wave-
lengths of 350 - 400 nm)
Combination of lasers
from Chapter 6.11
(VIS and UV lasers)
Combination of lasers from
Chapter 6.11 (VIS/UV lasers)
and Chapter 6.12 (IR lasers)
6.3 Laser class for VIS and UV systems
According to IEC/EN 60825-1, this laser scanning microscope is a laser device of Class IIIb.
3B / IIIb
3B / IIIb
4/IV
Avoidexposingeyesorskintodirectra­diation.
6.4 Laser class for MP systems
According to IEC/EN 60825-1, this laser scanning microscope is a laser device of
Class IV.
Avoidexposingeyesorskintodirectand indirect radiation.
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Safety Notes
6.5 Warnings, Safety Cautions, and Notes
Notes either contain additional information
on a specific topic or special instructions on the handling of the product.
A safety note points out an operation, a pro­cess,a condition oran instruction that must be observed strictly to prevent severe damage to the system or loss of data.
A laser warning points out an operation, a process, a condition or an instruction that must be observed strictly to prevent serious eye injuries to the persons using the system.
A high-voltage warning points out an ope­ration, a process, a condition or an instruc­tion that must be observed strictly to prevent possible injury or death of the persons using the system.
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Safety Notes
6.6 What should the user of the laser scanning mi­croscope observe?
The user of this product is responsible for proper and safe operation and safe maintenance of the system as well as for following all applicable safety regula­tions. The user is
fully liable for all consequences resulting from the use of the system for any other pur­poses than those listed in the Operating In­structions or the online help.
The user is obligated to perform and mo­nitor suitable safety measures
according to IEC / EN 60825-1 and
the corresponding national regulations.
Users must have received instructions concerning the risk potential associated with the operation of laser devices.
To assure classification as a 3B/IIIb or 4/IV laser product according to IEC/EN60825-1 and electrical safety com­pliance, all safety devices, interlocks, and safety systems of the laser device must be in operational condition.
Deactivating or damaging these safety devi­ces or any intervention in any of these safety devices may lead to serious eye injuries, physical injuries or property damages. In these cases, Leica Microsystems CMS GmbH does not assume any liability.
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According to IEC/EN 60825-1: ”Safety of laser products, Part 1: Equipment classification, requirements and user’s guide,” the user is required to
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Safety Notes
designate a Laser Safety Officer or a La­ser Protection Advisor.
Repairs and servicing may only be perfor­med by authorized Leica Microsystems CMS GmbH service personnel.
The user is fully liable for all consequences resulting from the use of the system if it is opened, improperly serviced or repaired by other persons than authorized Leica customer service representatives.
If repairs or service measures are perfor­med that require opening parts of the housing, only trained Leica service tech­nicians may occupy the room in which the laser scanning microscope is located.
Leica Micro­systems CMS GmbH will not be liable for da­mages resulting from nonobservance of the above information. The above information does not, in any way, implicitly or explicitly, modify the warranty and liability clauses con­tained in the general terms and conditions of Leica Microsystems CMS GmbH.
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