Leica TCS SP5 X User manual

Leica TCS SP5
User Manual

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Leica Microsystems CMS GmbH
Am Friedensplatz 3
68165 Mannheim
Germany
Telefon: [phone]
Fax: [phone]
E-Mail: [email protected]

Table of Contents
User Manual TCS SP5
V: 02 | Document-ID-No.:156500002
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.........................................
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................................

Table of Contents
User Manual TCS SP5
V: 02 | Document-ID-No.:156500002
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..........................

Table of Contents
User Manual TCS SP5
V: 02 | Document-ID-No.:156500002
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.......................................
11 Structure of the user interface page 85........
11.1 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 page 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 page 117.............................................

Table of Contents
User Manual TCS SP5
V: 02 | Document-ID-No.:156500002
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...................

Notes about this Manual
Page 1
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
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, inc-
luding but not limited to implicit warranties for
salability and for suitability for a specific pur-
pose. Leica Microsystems CMS GmbH is not
liable for errors or accidental damages or in-
cidental damages in conjunction with equip-
ment, performance or usage of this manual
or the examples contained therein.
©Leica Microsystems CMS GmbH
This document contains information that is
protected by copyright law. All rights reser-
ved. Duplication, adaptation or translation of
this manual requires the prior written approval
of Leica Microsystems CMS GmbH unless
otherwise permitted through copyright arran-
gements.
The programs for controlling this product are
protected by copyright law. All rights reser-
ved. Reproduction, adaptation or translation
of these programs without prior written ap-
proval of Leica Microsystems CMS GmbH is
strictly prohibited.
MicrosoftWindows,WindowsXP,theWin-
dows logo and the Windows CE logo are ei-

Notes about this Manual
Page 2 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
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.

General
Page 3
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
2 General
2.1 About these operating instructions
The main focus of these operating instruc-
tions is directed to the safety instructions that
must be observed while working with the la-
ser scanning microscope.
In addition, these operating instructions pro-
vide a rough overview of the operating princi-
ple of laser scanning microscopes. It pre-
sents you with the first steps for starting and
startup of the system and provides a descrip-
tion of the Leica Application Suite Advanced
Fluorescence (LAS AF).
The Leica TCS SP5 is supplied with the la-
test 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 instruc-
tions. 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 instruc-
tions about the corresponding software func-
tions.
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 spe-
cific functions can subsequently be obtained
electronically at your TCS workstation.

General
Page 4 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002

Proper Intended Use
Page 5
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
3 Proper intended use
The system was designed for confocal recor-
ding (laser scanning images) of fluore-
scence-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 out-
side the specifications of Leica Microsystems
CMSGmbHaswellasanyrisksresulting
from it. In such cases, the declaration of con-
formity becomes invalid.

Proper Intended Use
Page 6 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002

Legal Notes
Page 7
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
4LegalNotes
Manufactured in the Federal Republic of
Germany.
© Copyright 2005,
Leica Microsystems CMS GmbH.
All rights reserved.
No part of this publication may be reprodu-
ced or transmitted in any form or by any
means, electronic or mechanical, including
photocopying, recording, or storing in a re-
trieval system, or translating into any lan-
guage in any form without the express writ-
ten permission of Leica Microsystems CMS
GmbH. This also includes photocopying, re-
cording 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 be-
nefit of safe, efficient operation and mainte-
nance 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 pro-
hibited, and delivery of this document shall
not constitute any license or implied authori-
zation to do so.

Legal Notes
Page 8 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
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 wi-
thout 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, inclu-
ding 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 con-
sequential damages that could result from
the use of the material described in this do-
cument.
4.4 Patents
The product is protected by the following US
patents:
5,886,784; 5,903,688; 6,137,627; 6,222,961;

Legal Notes
Page 9
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
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.

Legal Notes
Page 10 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
4.5 TRADEMARKS
Throughout this manual, trademarked names
may be used. Rather than including a trade-
mark (™) 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.
4.6 Software licenses
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.

Specifications
Page 11
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
5 TCS SP5 specifications
5.1 System overview
12
3
4
5
6
figure1. System components (overview)
1TCS SP5 Scanner
2Control panel
3TCS workstation
4Supply unit
5Panel box
6Stand with scanner

Specifications
Page 12 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
5.2 Dimensions
5.2.1 System with inverted stand:
figure2. Dimensions of the TCS SP5 with inverted stand

Specifications
Page 13
User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
5.2.2 System with upright stand
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 environment is as dust-
free as possible.

Specifications
Page 14 User Manual TCS SP5
V: 02 | Document-ID-No.: 156500002
Please also read the notes on protection
against dust in the chapters on maintenance
and cleaning. Installing the system in darke-
ned rooms is also advisable.
For installation, maintenance and trans-
port, 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 ap-
ply 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% (non-
condensing)
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
Power consumption: 2 x 1.6kW
Overvoltage category: II
The building installation must feature two se-
parate power connections with the following
fuse protection:
D100 V - 120 V AC power: 20 A