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Cell Separations by Flow Cytometry

  • Protocol
Metastasis Research Protocols

Part of the book series: Methods in Molecular Medicine ((MIMM,volume 58))

Abstract

Flow cytometry is a means of measuring the physical and chemical characteristics of particles in a fluid stream as they pass one by one past a sensing point. The modern flow cytometer consists of a light source, collection optics and detectors, and a computer to translate signals into data. In effect, a flow cytometer can be described as a large and powerful fluorescence microscope in which the light source is of a highly specific wavelength, generally produced by a laser, and the human observer is replaced by a series of optical filters and detectors that aim to make the instrument more objective and more quantitative. As a cell passes through the laser beam, light is scattered in all directions, and also at this point any fluorochromes present on the cell are excited and emit light of a higher wavelength. Scattered and emitted light is collected by two lenses-one set in front of the light source and one set at right angles to it. By a series of beam splitters, optical filters, and detectors the wavelengths of light specific for particular fluorochromes can be isolated and quantitated-up to six fluorochromes can be measured in some flow cytometers. A simplified diagram of the optical setup for two-color analysis is shown in Fig. 1. The theory of operation of flow cytometers is well documented, and there are several good general books on the subject (1-3).

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Author information

Authors and Affiliations

  1. FACS Laboratory, Imperial Cancer Research Fund, London, UK

    Derek Davies

Authors
  1. Derek Davies
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Editor information

Editors and Affiliations

  1. Research School of Biological and Molecular Sciences, Oxford Brookes University, Oxford, UK

    Susan A. Brooks

  2. Institute for Anatomy, Department of Neuroanatomy, University Hospital Hamburg-Eppendorf, Hamburg, Germany

    Udo Schumacher

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© 2001 Humana Press Inc.

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Davies, D. (2001). Cell Separations by Flow Cytometry. In: Brooks, S.A., Schumacher, U. (eds) Metastasis Research Protocols. Methods in Molecular Medicine, vol 58. Humana, Totowa, NJ. https://doi.org/10.1385/1-59259-137-X:003

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  • DOI: https://doi.org/10.1385/1-59259-137-X:003

  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-0-89603-615-4

  • Online ISBN: 978-1-59259-137-4

  • eBook Packages: Springer Protocols

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