TripleFRET measurements in flow cytometry

Authors

  • Ákos Fábián,

    1. Department of Biophysics and Cell Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary
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    • Á. Fábián and G. Horváth authors contributed equally to the article.

  • Gábor Horváth,

    1. Institute of Innate Immunity, University Hospitals, University of Bonn, Germany
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    • Á. Fábián and G. Horváth authors contributed equally to the article.

  • György Vámosi,

    1. Department of Biophysics and Cell Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary
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  • György Vereb,

    1. Department of Biophysics and Cell Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary
    2. MTA-DE Cell Biology and Signaling Research Group, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary
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  • János Szöllősi

    Corresponding author
    1. Department of Biophysics and Cell Biology, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary
    2. MTA-DE Cell Biology and Signaling Research Group, Research Center for Molecular Medicine, Medical and Health Science Center, University of Debrecen, Hungary
    • Department of Biophysics and Cell Biology, Faculty of Medicine, Medical and Health Science Center, University of Debrecen, P.O.Box 39, Nagyerdei krt. 98, H-4012 Debrecen, Hungary
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Errata

This article is corrected by:

  1. Errata: TripleFRET measurements in flow cytometry Volume 83A, Issue 7, 672, Article first published online: 21 May 2013

Abstract

A frequently used method for viewing protein interactions and conformation, Förster (fluorescence) resonance energy transfer (FRET), has traditionally been restricted to two fluorophores. Lately, several methods have been introduced to expand FRET methods to three species. We present a method that allows the determination of FRET efficiency in three-dye systems on a flow cytometer. TripleFRET accurately reproduces energy transfer efficiency values measured in two-dye systems, and it can indicate the presence of trimeric complexes, which is not possible with conventional FRET methods. We also discuss the interpretation of energy transfer values obtained with tripleFRET in relation to spatial distribution of labeled molecules, specifically addressing the limitations of using total energy transfer to determine molecular distance. © 2013 International Society for Advancement of Cytometry

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