BD LSRII Operation instructions

BD Biosciences
2350 Qume Drive
San Jose, CA
95131-1807
Draft Rev A
September 2002
BD LSRII Overview
Workbook

ii BD LSRII Overview Workbook Draft

iii Draft Copyright
BD Biosciences • 2350 Qume Drive • San Jose, California
• 95131-1807
© 2002 Becton, Dickinson and Company. All rights reserved. No part of
this publication may be reproduced, transmitted, transcribed, stored in
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BD Biosciences, 2350 Qume Drive, San Jose, CA 95131, United States of
America.
BD Biosciences reserves the right to change its products and services at
any time to incorporate the latest technological developments. Although
this guide has been prepared with every precaution to ensure accuracy,
BD Biosciences assumes no liability for any errors or omissions, nor for
any damages resulting from the application or use of this information.
This workbook is subject to change without notice. BD Biosciences
welcomes customer input on corrections and suggestions for
improvement.
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For Research Use Only. Not for use in diagnostic or therapeutic
procedures.

iv BD LSRII Overview Workbook Draft

v Draft Table of Contents
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Table of Contents
1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1
About the Instrument . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Power Switch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4
Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4
Optics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9
Electronics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-14
Software . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-14
Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-15
2 Operations and Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1
Lab Exercise: BD LSRII Startup . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3
Lab Exercise: BD LSRII Shutdown . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-7
BD LSRII Maintenance and Care Procedures . . . . . . . . . . . . . . . . . . . . . . . . 2-9
Regular Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-9
Periodic Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-13
Laser Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-18
3 Optics and Lasers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1
Lasers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3
Optical Filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4
Optical Filters in the BD LSRII . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4
4 Instrument Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-1
Instrument Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Lab Exercise: Performing Quality Control . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Preparing the QC Sample . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Appendix A Worksheets . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-1
Appendix B Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1
BD LSR Overview Course Evaluation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . E-1

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Draft
1
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•
Introduction
After completing this module, you will be able to:
◆
describe the three systems of a flow cytometer
◆
describe the anatomy BD LSRII flow cytometer

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1-3 Draft Module 1: Introduction
About the Instrument
The BD LSRII is a ten-color benchtop flow cytometer. It combines ease of
use with versatility and performance for advanced research applications.
Figure 1-a BD LSRII flow cytometer with outer covering removed
Power Switch
The power switch is located on the lower-right side of the BD LSRII
instrument.

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Control Panel
The fluidics controls, which consists of six buttons and one knob, are
found on the control panel in the front of the instrument.
Figure 1-b BD LSRII Control Panel
Fluidics
The purpose of the fluidics system is to carry the sample out of the sample
tube and into the sensing region of the flow cell. Cells are carried in the
sample core stream in single file and measured individually.
The fluidics system in the BD LSRII flow cytometer is pressure driven, a
built-in air pump provides a sheath pressure of 6.0 psi. After passing
through the sheath filter, sheath fluid is introduced into the lower chamber
of the quartz flow cell.
The sample to be analyzed arrives in a separate pressurized stream. When
a sample tube is placed on the sample injection port (SIP), the sample is
forced up and injected into the lower chamber of the flow cell by a slight
overpressure relative to the sheath fluid. The conical shape of the lower
chamber creates a laminar sheath flow that carries the sample core
upward through the center of the flow cell, where the particles to be