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Single-Molecule Fluorescence Coincidence Spectroscopy and its Application to Resonance Energy Transfer

Authors

  • Dr. Angel Orte,

    Corresponding author
    1. Department of Physical Chemistry. Faculty of Pharmacy. University of Granada. Campus Cartuja, 18071, Granada (Spain), Fax: (+34)-958-244090
    • Department of Physical Chemistry. Faculty of Pharmacy. University of Granada. Campus Cartuja, 18071, Granada (Spain), Fax: (+34)-958-244090
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  • Dr. Richard W. Clarke,

    1. Department of Chemistry, University of Cambridge, Lensfield road, Cambridge, CB2 1EW (UK), Fax: (+44)-1223-336362
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  • Prof. David Klenerman

    Corresponding author
    1. Department of Chemistry, University of Cambridge, Lensfield road, Cambridge, CB2 1EW (UK), Fax: (+44)-1223-336362
    • Department of Chemistry, University of Cambridge, Lensfield road, Cambridge, CB2 1EW (UK), Fax: (+44)-1223-336362
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Abstract

The use of Förster resonance energy transfer (FRET) as a tool to study biomolecules has been greatly enhanced by new advances in single-molecule fluorescence (SMF) techniques. This has allowed new insights into the structure and dynamics of complex biomolecular machinery. However, there are still technical drawbacks in the application of conventional SMF–FRET. Herein, we review the use of single-molecule coincidence spectroscopy to study FRET systems, an analytical variation of the conventional scheme, using one or two confocal lasers of different colours. We highlight the advantages of the coincidence spectroscopy and illustrate this with examples of its application to some biological systems of interest.

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