TripleFRET measurements in flow cytometry

Ákos Fábián, Gábor Horváth, György Vámosi, György Vereb, János Szöllosi

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10 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)375-385
Number of pages11
JournalCytometry Part A
Volume83 A
Issue number4
DOIs
Publication statusPublished - Apr 1 2013

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Keywords

  • Förster (fluorescence) resonance energy transfer
  • Relay transfer
  • Trimeric complexes
  • TripleFRET

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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