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Fluorescence fluctuation spectroscopy: ushering in a new age of enlightenment for cellular dynamics

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Abstract

Originally developed for applications in physics and physical chemistry, fluorescence fluctuation spectroscopy is becoming widely used in cell biology. This review traces the development of the method and describes some of the more important applications. Specifically, the methods discussed include fluorescence correlation spectroscopy (FCS), scanning FCS, dual color cross-correlation FCS, the photon counting histogram and fluorescence intensity distribution analysis approaches, the raster scanning image correlation spectroscopy method, and the Number and Brightness technique. The physical principles underlying these approaches will be delineated, and each of the methods will be illustrated using examples from the literature.

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Acknowledgments

The authors wish to thank Nicholas James for critically reading the manuscript. This work was supported by National Institutes of Health grant RO1GM076665 (DMJ) and a grant from Allergan, Inc.

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  1. Department of Cell and Molecular Biology, John A. Burns School of Medicine, University of Hawaii, 651 Ilalo St., BSB 222, Honolulu, HI, 96813, USA

    David M. Jameson & Justin A. Ross

  2. Department of Pharmacology, U.T. Southwestern Medical Center, 6001 Forest Park, Dallas, TX, 75390-9041, USA

    Joseph P. Albanesi

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Correspondence to David M. Jameson.

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Jameson, D.M., Ross, J.A. & Albanesi, J.P. Fluorescence fluctuation spectroscopy: ushering in a new age of enlightenment for cellular dynamics. Biophys Rev 1, 105–118 (2009). https://doi.org/10.1007/s12551-009-0013-8

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