Fluorescence correlation spectroscopy for the detection and study of single molecules in biology

Authors

  • Miguel Ángel Medina,

    Corresponding author
    1. Department of Molecular Biology and Biochemistry, Faculty of Sciences, University of Málaga, Spain
    • Department of Molecular Biology and Biochemistry, Faculty of Sciences, University of Málaga, E-29071 Málaga, Spain.
    Search for more papers by this author
  • Petra Schwille

    1. Experimental Biophysics Group, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany
    Search for more papers by this author

Abstract

The recent development of single molecule detection techniques has opened new horizons for the study of individual macromolecules under physiological conditions. Conformational subpopulations, internal dynamics and activity of single biomolecules, parameters that have so far been hidden in large ensemble averages, are now being unveiled. Herein, we review a particular attractive solution-based single molecule technique, fluorescence correlation spectroscopy (FCS). This time-averaging fluctuation analysis which is usually performed in Confocal setups combines maximum sensitivity with high statistical confidence. FCS has proven to be a very versatile and powerful tool for detection and temporal investigation of biomolecules at ultralow concentrations on surfaces, in solution, and in living cells. The introduction of dual-color cross-correlation and two-photon excitation in FCS experiments is currently increasing the number of promising applications of FCS to biological research. BioEssays 24:758–764, 2002. © 2002 Wiley Periodicals, Inc.

Ancillary