Light Scatter: Detection and Usage

Gary C. Salzman1

1 Los Alamos National Laboratory, Los Alamos, New Mexico
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 1.13
DOI:  10.1002/0471142956.cy0113s09
Online Posting Date:  May, 2001
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Light scatter is one of the most basic of measurements made in flow cytometry. This unit presents a short commentary on what light scatter is and how it is measured. First, the author describes the basic principles of the scattering of light from small particles such as cells. This is followed by a discussion of methods for detecting scattered light in a flow cytometer. The unit ends with a brief survey showing how light scattering is used in a broad range of flow cytometry applications.

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Table of Contents

  • Light Waves
  • Scattering
  • Scattering from a Dipole
  • Scattering from Several and Many Dipoles
  • Scattering by Any Other Name
  • Forward Scatter Detection
  • Side Scatter Detection
  • Combined Forward and Side Scatter Detection
  • Conclusion
  • Literature Cited
  • Figures
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Literature Cited

Literature Cited
   Barber, P.W., Hill, S.C. 1990. Light Scattering by Particles Computational Methods. Advanced Series in Applied Physics Vol. 3 World Scientific Publishing Corp., London.
  This work was performed under the auspices of the National Flow Cytometry Resource at Los Alamos National Laboratory and was supported by the U.S. Army Soldier Biological and Chemical Command, Aberdeen Proving Ground, Md. The author would like to thank Georgia Farris (Los Alamos) for collecting FACSCalibur data and Jianhong Wang (BDIS) and Robert Hoffman (BDIS) for providing optical data for the FACSCalibur.
   Bohren, C.F. 1987. Physics on a manure heap: More about black clouds. In Clouds in a Glass of Beer, pp.139‐142. John Wiley & Sons, New York.
   Bohren, C.F., Huffman, .R. 1983. Absorption and Scattering of Light by Small Particles. John Wiley & Sons, New York
   Doornbos, R.M.P., Hoekstra, A.G., Deurloo, K.E.I., De Grooth, B.G., Sloot, P.M.A., and Greve, J. 1994. Lissajous like patterns in scatter plots of calibration beads. Cytometry 16:236‐242.
   Hoffman, R.A., Kung, P.C., Hansen, W.P., and Goldstein, G. 1980. Simple and rapid measurement of human T lymphocytes and their subclasses in peripheral blood. Proc. Natl. Acad. Sci. U.S.A 77:4914‐4917.
   Lorenz, L.V. 1890. Upon the light reflected and refracted by a transparent sphere. Vidensk. Selsk. Skrifler 6:1‐62.
   Mie, G. 1908. Considerations on the optics of turbid media, especially colloidal metal sols. Ann. Physik 25:377‐442.
   Salzman, G.C., Crowell, J.M., Martin, J.C., Trujillo, T.T., Romero, A., Mullaney, P.F., and LaBauve, P.M. 1975. Cell classification by laser light scattering. Identification and separation of unstained leukocytes. Acta. Cytol 19:374‐377.
   Salzman, G.C., Singham, S.B., Johnston, R.G., and Bohren, C.F. 1990. Light scattering and cytometry. In Flow Cytometry and Sorting (T. Lindmo eds.) 2 ed. pp. 81‐107. Wiley‐Liss, New York.
   Strutt, J.W. (Lord Rayleigh). 1871. On the light from the sky, its polarization and colour. Phil. Mag 41:107‐120.
   Van de Hulst, H.C. 1981. Light Scattering by Small Particles. Dover Publications, Inc. New York.
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