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introduction to flow cytometry--- a learning guide

introduction to flow cytometry--- a learning guide

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 Introduction toFlow Cytometry:
 A Learning Guide
 11-11032-03 rev. ADecember 2002
 
 Introduction to Flow Cytometry: A Learning Guide 
 Copyright 
 © 2002 Becton, Dickinson and Company. All rights reserved. No part of this publication may be reproduced,transmitted, transcribed, stored in retrieval systems, or translated into any language or computer language, in any form or by any means: electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without the prior written permission of BD Biosciences, 2350 Qume Drive, San Jose, CA 95131, United States of America.
 Disclaimer
 BD Biosciences reserves the right to change its products and services at any time to incorporate the latesttechnological developments. This guide is subject to change without notice. BD Biosciences welcomes customerinput on corrections and suggestions for improvement. Although this guide has been prepared with every precaution to ensure accuracy, BD Biosciences assumes noliability for any errors or omissions, nor for any damages resulting from the application or use of this information.
 Trademarks
  Apple, the Apple logo, Mac, Macintosh, and Power Macintosh are trademarks of Apple Computer, Inc., registeredin the U.S. and other countries. Finder is a trademark of Apple Computer, Inc.Modfit
L
 and QuantiCALC are trademarks of Verity Software House, Inc.BD Attractors, BD CellQuest, BD FACS, BD FACSCalibur, BD FACSDiVa, BD FACScan, BD FACSort,BD FACSVantage, BD FACSVantage SE, BD MultiSET, are trademarks of Becton, Dickinson and Company.
 
 iii
 Introduction to Flow Cytometry: A Learning Guide
 
Table of Contents
 Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vFor More Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vChapter 1 Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
 Review Questions: Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
 Chapter 2 Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
 Review Questions: Fluidics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8
 Chapter
3
Generation of Scatter and Fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . 93.1
Light Scatter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
 Review Questions: Light Scatter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
 3.2
Fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
 Review Questions: Fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
 Chapter
4 Optical System
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 174.1
Optical Bench . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17Review Questions: Optical Bench . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
 4.2
Optical Filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20Review Questions: Optical Filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
 4.3
Signal Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22Review Questions: Signal Detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24
 4.4
Threshold . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25Review Questions: Threshold . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25
 Chapter
5 Data Analysis
. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 275.1
Data Collection and Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27Review Questions: Data Collection and Display . . . . . . . . . . . . . . . . . . . . . 28
 5.2
Gating . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29Review Questions: Gating . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
 5.3 Data Analysis for Subsetting Applications . . . . . . . . . . . . . . . . . . . . . . . . . 305.4 Data Analysis for Other Applications . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34
 Review Questions: Data Analysis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

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