Leica TCS SP5 X User manual

Leica TCS SP5
User Manual

Published by:
Leica Microsystems CMS GmbH
Am Friedensplatz 3
D-68165 Mannheim (Germany)
http://www.leica-microsystems.com
http://www.confocal-microscopy.com
Responsible for contents: Leica Microsystems CMS GmbH
Copyright © Leica Microsystems CMS GmbH.
All rights reserved.
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Table of Contents
1. Notes about this manual..................................................................................... 7
1.1 Copyright ............................................................................................................ 7
1.2 Patents ............................................................................................................... 8
2. General ................................................................................................................. 9
2.1 About these operating instructions ..................................................................... 9
2.2 Intended use....................................................................................................... 9
3. TCS SP5 specifications .................................................................................... 11
3.1 System overview .............................................................................................. 11
3.2 Dimensions....................................................................................................... 12
3.2.1 System with inverted stand:............................................................................ 12
3.2.2 System with upright stand............................................................................... 12
3.3 Requirements regards ambient conditions ....................................................... 13
3.4 Permissible ambient conditions ........................................................................ 13
3.5 Electrical connections....................................................................................... 14
3.6 Waste heat/required cooling performance........................................................ 14
3.7 Required safety measures for VIS and UV systems......................................... 14
3.8 Required laser safety measures for MP systems ............................................. 14
4. Safety instructions and their meaning ............................................................ 15
5. Safety instructions ............................................................................................ 17
5.1 Which standards does this product meet?........................................................ 17
5.2 Which laser class does the product have? ....................................................... 18
5.3 Laser class for VIS and UV systems ................................................................ 18
5.4 Laser class for MP systems.............................................................................. 19
5.5 What should the user of the laser scanning microscope observe? .................. 19
5.6 Safety Notes for the User ................................................................................. 20
5.7 Safety Notes for Operation ............................................................................... 21
5.8 Specific Safety Notes ....................................................................................... 23
5.8.1 Operational reliability ...................................................................................... 23
5.8.2 De-energizing the system ............................................................................... 24
5.8.3 Maximum current load of the power outlet strip at the supply unit.................. 24
5.9 Laser safety devices......................................................................................... 25
5.9.1 Eye protection for VIS or UV systems ............................................................ 25
5.9.2 Detachable-key switch.................................................................................... 25
5.9.3 Emissions warning indicators ......................................................................... 26
5.9.4 Remote interlock connector on the supply unit............................................... 27
5.9.5 Remote interlock connectors on additional external lasers............................. 28
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5.9.6 Interlock connection on scanner ..................................................................... 28
5.9.7 Function and position of safety switches ........................................................ 29
5.9.8 Transmitted-light lamp housing for inverted stands ........................................ 30
5.9.9 Transmitted-light lamp housing for upright stands.......................................... 31
5.9.10 Mirror housing on upright stand ...................................................................... 32
5.9.11 Special laser safety equipment....................................................................... 34
5.9.11.1 Safety beam guide and beam collector...................................................... 34
5.9.11.2 Eye protection for MP systems .................................................................. 35
5.9.11.3 Shielding in MP systems (IR lasers) .......................................................... 35
5.10 Overview of usable VIS/UV lasers.................................................................... 36
5.11 Overview of usable MP lasers (IR lasers)......................................................... 37
5.12 Safety label on TCS SP5 system ..................................................................... 37
5.12.1 Inverted stand DMI 6000 CS .......................................................................... 38
5.12.2 Upright stand DM 5000/6000 CS.................................................................... 40
5.12.3 Scan head....................................................................................................... 42
5.12.4 Supply unit ...................................................................................................... 43
5.12.5 MP beam coupling unit ................................................................................... 44
5.12.6 Cover (for replacement flange) ....................................................................... 45
5.12.7 Mirror Housing ................................................................................................ 46
5.13 Installation and storage site requirements ........................................................ 47
5.14 Changing the installation site............................................................................ 47
5.15 Scanner cooling................................................................................................ 48
6. Care and Maintenance ...................................................................................... 49
6.1 Cleaning the optical system of the microscope ................................................ 49
6.2 Cleaning the microscope surface ..................................................................... 50
7. Startup of the system........................................................................................ 51
7.1 Starting and logging on..................................................................................... 51
7.2 Setting Up Users .............................................................................................. 52
7.3 Starting up ........................................................................................................ 53
7.4 Starting the LAS AF.......................................................................................... 55
8. Introduction to LAS AF ..................................................................................... 57
8.1 General............................................................................................................. 57
8.2 Online help ....................................................................................................... 57
8.2.1 Structure of the online help............................................................................. 57
8.2.2 Calling Online Help ......................................................................................... 58
8.2.3 Full-text search with logically connected search terms................................... 59
8.3 Structure of the user interface .......................................................................... 60
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8.3.1 General structure of the user interface ........................................................... 60
8.4 Key combinations ............................................................................................. 61
9. Introduction to Confocal Work......................................................................... 63
9.1 Preparation ....................................................................................................... 63
9.1.1 The objective .................................................................................................. 63
9.1.2 Conventional microscopy................................................................................ 64
9.1.3 Why scan? ...................................................................................................... 67
9.1.4 How is an optical section created? ................................................................. 68
9.2 Acquiring optical sections ................................................................................. 70
9.2.1 Data acquisition .............................................................................................. 70
9.2.2 Illumination...................................................................................................... 72
9.2.3 Beam splitting ................................................................................................. 73
9.2.4 Emission bands .............................................................................................. 74
9.2.5 The pinhole and its effects.............................................................................. 76
9.2.6 Image detail and raster settings...................................................................... 77
9.2.7 Signal and noise ............................................................................................. 81
9.2.8 Profile cuts ...................................................................................................... 83
9.3 Multiparameter fluorescence ............................................................................ 84
9.3.1 Illumination...................................................................................................... 84
9.3.2 Beam splitting ................................................................................................. 86
9.3.3 Emission bands .............................................................................................. 86
9.3.4 Crosstalk......................................................................................................... 86
9.3.5 Sequential capture.......................................................................................... 87
9.3.6 Unmixing......................................................................................................... 87
9.4 3D Series.......................................................................................................... 88
9.4.1 z-stack ............................................................................................................ 88
9.4.2 Layer thicknesses ........................................................................................... 88
9.4.3 Distances ........................................................................................................ 89
9.4.4 Data volumes.................................................................................................. 89
9.4.5 Depictions ....................................................................................................... 90
9.4.5.1 Gallery........................................................................................................ 90
9.4.5.2 Movie.......................................................................................................... 90
9.4.5.3 Orthogonal projections ............................................................................... 91
9.4.5.4 Rotated projections .................................................................................... 92
9.5 Time series ....................................................................................................... 92
9.5.1 scan speed ..................................................................................................... 93
9.5.2 Points.............................................................................................................. 93
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9.5.3 Lines ............................................................................................................... 93
9.5.4 Planes............................................................................................................. 93
9.5.5 Spaces (time-space)....................................................................................... 93
9.5.6 FRAP measurements ..................................................................................... 94
9.6 Spectral series.................................................................................................. 94
9.6.1 Data acquisition and utilization ....................................................................... 94
9.6.2 About spectral resolution ................................................................................ 94
9.7 Combinatorial analysis ..................................................................................... 95
10. Switching off the system .................................................................................. 97
11. Disposal ............................................................................................................. 99
12. Contact ............................................................................................................. 101
13. Declaration of conformity............................................................................... 103
14. Glossary ........................................................................................................... 105
15. Safety data sheets........................................................................................... 111
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1. Notes about this manual
1.1 Copyright
The instructions contained in the following documentation reflect state-of-the-art technology
and knowledge standards. We have compiled the texts and illustrations as accurately as
possible. Nevertheless, no liability of any kind may be assumed for the accuracy of this
manual's contents. If you have any comments on these operating instructions or on any of
our other documentation we would be pleased to receive them. The information in this
manual is subject to modification at any time and without notification.
All rights to this documentation are held by Leica Microsystems CMS GmbH. Adaptation,
translation and reproduction of text or illustrations (in whole or in part) by print, photocopy,
microfilm or other method (including electronic systems) is not allowed without express
written permission from Leica Microsystems CMS GmbH.
Programs such as LAS and LAS AF are protected by copyright laws. All rights reserved.
Reproduction, adaptation or translation of these programs is prohibited without prior written
permission from Leica Microsystems CMS GmbH.
The term "Windows" may be used in the following text without further identification. It is a
registered trademark of the Microsoft Corporation. Otherwise, no inference with regard to the
free usability of product names may be drawn from the use of those names. All other brand
names and product names in this document are brands, service marks, trademarks or
registered trademarks of the respective manufacturers.
Manufactured in the Federal Republic of Germany.
© Copyright Leica Microsystems CMS GmbH.
All rights reserved.
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1.2 Patents
The product is protected under the following US patents:
5,886,784; 5,903,688; 6,137,627; 6,222,961; 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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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 software Leica Application Suite
Advanced Fluorescence (LAS AF).
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 intentionally 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. Please refer to the online help for further information on particular functions.
2.2 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 CMS GmbH as well as any risks resulting from it. In
such cases, the declaration of conformity becomes invalid.
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