Lasers for Flow Cytometry

Howard M. Shapiro1, William G. Telford2

1 The Center for Microbial Cytometry, West Newton, Massachusetts, 2 Experimental Transplantation and Immunology Branch, National Cancer Institute, Bethesda, Maryland
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 1.9
DOI:  10.1002/0471142956.cy0109s49
Online Posting Date:  July, 2009
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Abstract

Lasers are the principal light sources for flow cytometers. Most modern benchtop instruments feature air‐cooled lasers emitting 10 to 25 mW of light at 488 nm. This unit covers the various types of lasers available and the qualities that make them suitable or unsuitable for use in flow cytometers. Also included is a discussion of future directions, particularly in the development of solid‐state devices. Curr. Protoc. Cytom. 49:1.9.1‐1.9.17. © 2009 by John Wiley & Sons, Inc.

Keywords: flow cytometry; Gaussian beams; illumination optics; lasers

     
 
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Table of Contents

  • Introduction
  • Why Lasers or Arc Lamps are Needed as Light Sources for Flow Cytometry
  • How Lasers Work
  • Lasers Used in Flow Cytometry
  • Laser Safety
  • Future Directions
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Figures

  •   FigureFigure 1.9.1 Transitions between electronic energy states in a typical laser.
  •   FigureFigure 1.9.2 Schematic diagram of a laser. Brewster windows used to produce polarized output would be placed between the medium and the mirrors; a Littrow prism or wedge used for wavelength selection is typically placed between the medium and the high reflector mirror.

Videos

Literature Cited

   Kapoor, V., Subach, F.V., Kozlov, V.G., Grudinin, A., Verkhusha, V.V., and Telford, W.G. 2007. New lasers for flow cytometry: Filling the gaps. Nat. Methods 4:678‐679.
   Telford, W.G., Bradford, J., Godfrey, W., Robey, R.W., and Bates, S.E. 2007. Side population analysis using a violet‐excited cell‐permeable DNA binding dye. Stem Cells 25:1029‐1036.
Key References
   Harbison, J.P. and Nahory, R.E. 1997. Lasers: Harnessing the Atom's Light. Scientific American Library, New York.
  Aimed at the interested layman, this book is beautifully illustrated and includes a detailed discussion of the operation of semiconductor lasers.
   Hecht, J. 1992. The Laser Guidebook, 2nd ed. Tab Books (McGraw‐Hill), Blue Ridge Summit, Pa.
  Hecht's books provide substantial technical detail about lasers in a manner accessible to readers without a strong background in physics and engineering.
   Hecht, J. 2008. Understanding Lasers: An Entry‐Level Guide, 3rd ed. Wiley‐IEEE Press, New York.
  First description of the use of a supercontinuum laser in flow cytometry.
   Kapoor et al., 2007. See above.
  This book provides more particulars about specific flow cytometric applications of various lasers than appear here.
   Shapiro, H.M. 2003. Practical Flow Cytometry, 4th ed. Wiley‐Liss, Hoboken, N.J.
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