Fluorescent Proteins for Flow Cytometry

Teresa S. Hawley1, Robert G. Hawley2, William G. Telford3

1 Flow Cytometry Core Facility, Laboratory of Genome Integrity, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, 2 Department of Anatomy and Regenerative Biology, George Washington University School of Medicine and Health Sciences, Washington, D.C., 3 Flow Cytometry Core Facility, Experimental Transplantation and Immunology Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda
Publication Name:  Current Protocols in Cytometry
Unit Number:  Unit 9.12
DOI:  10.1002/cpcy.17
Online Posting Date:  April, 2017
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Abstract

Fluorescent proteins have become standard tools for cell and molecular biologists. The color palette of fluorescent proteins spans the ultraviolet, visible, and near‐infrared spectrum. Utility of fluorescent proteins has been greatly facilitated by the availability of compact and affordable solid state lasers capable of providing various excitation wavelengths. In theory, the plethora of fluorescent proteins and lasers make it easy to detect multiple fluorescent proteins simultaneously. However, in practice, heavy spectral overlap due to broad excitation and emission spectra presents a challenge. In conventional flow cytometry, careful selection of excitation wavelengths and detection filters is necessary. Spectral flow cytometry, an emerging methodology that is not confined by the “one color, one detector” paradigm, shows promise in the facile detection of multiple fluorescent proteins. This chapter provides a synopsis of fluorescent protein development, a list of commonly used fluorescent proteins, some practical considerations and strategies for detection, and examples of applications. © 2017 by John Wiley & Sons, Inc.

Keywords: fluorescent proteins; conventional flow cytometry; spectral flow cytometry; lasers

     
 
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Table of Contents

  • Introduction
  • Flow Cytometry of Fluorescent Proteins
  • Lasers for Fluorescent Proteins
  • Examples of Fluorescent Protein Applications
  • Conclusion
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Literature Cited

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