FRET and FCS - Friends or foes?

Research output: Contribution to journalShort survey/reviewContributedpeer-review

Contributors

  • Harekrushna Sahoo - , TUD Dresden University of Technology (Author)
  • Petra Schwille - , Chair of Biophysics (Author)

Abstract

Fluorescence correlation spectroscopy (FCS) and Förster resonance energy transfer (FRET) are both scientific concepts that are frequently discussed in the context of single-molecule fluorescence techniques. In contrast to FCS, FRET is strictly not a technique but a photophysical phenomenon, which can be employed in combination with any method that probes fluorescence intensity or lifetime. Thus, the combination of FCS with FRET is possible and - although these concepts are quite often treated as alternative approaches, particularly for the analysis of biological systems - also quite attractive. However, under certain circumstances, for example, for applications of fluorescence cross-correlation spectroscopy, FRET effects can cause significant complications for quantitative data analysis, and careful calibration has to be carried out to avoid FRET-induced artifacts. This can be most elegantly done if alternating excitation schemes such as PIE (pulsed interleaved excitation) are employed. In this minireview, we discuss the potential and the caveats of FCS combined with FRET and give a short record on successful and promising applications.

Details

Original languageEnglish
Pages (from-to)532-541
Number of pages10
JournalChemPhysChem
Volume12
Issue number3
Publication statusPublished - 25 Feb 2011
Peer-reviewedYes

Keywords

Keywords

  • Fluorescence correlation spectroscopy, Fluorophore labeling, FRET, Photophysics, Single-molecule studies