BD LSRII Instructions de fonctionnement

BD Biosciences

2350 Qume Drive

San Jose, CA

95131-1807

Draft Rev A

September 2002

BD LSRII Overview

Workbook

BD LSRII Overview Workbook Draft

iii

Draft Copyright

BD Biosciences • 2350 Qume Drive • San Jose, California

• 95131-1807

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BD Biosciences, 2350 Qume Drive, San Jose, CA 95131, United States of

America.

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any time to incorporate the latest technological developments. Although

this guide has been prepared with every precaution to ensure accuracy,

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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.

BD LSRII Overview Workbook Draft

Draft Table of Contents

•

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

vi BD LSRII Overview Workbook Draft

Draft

•

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

1-2

BD LSRII Overview Workbook Draft

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.

1-4 BD LSRII Overview Workbook Draft

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

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