Research Techniques Made Simple: Methodology and Applications of Förster Resonance Energy Transfer (FRET) Microscopy

Joshua A. Broussard, Kathleen J. Green*

*Corresponding author for this work

Research output: Contribution to journalShort surveypeer-review

12 Scopus citations

Abstract

Classical biochemical techniques have contributed a great deal to our understanding of the mechanisms regulating fundamental biological processes. However, these approaches are typically end-point, population-based assays and are often insufficient in examining transient molecular events. Förster resonance energy transfer (FRET) microscopy is a powerful technique capable of investigating dynamic interactions between proteins and a plethora of biochemical signaling events based on the development of specific biosensors. This technique exploits the principle that when FRET occurs, energy from a donor fluorophore is transferred to an acceptor fluorophore only when certain conditions are met. These include dependence on both distance and fluorophore orientation. In this article, applications of FRET microscopy to protein interactions and modifications are discussed, and examples are given of the types of biosensors that can be developed. There are a number of methods to measure FRET. The most common modalities and specific advantages and shortcomings for each are reviewed. Finally, general considerations and guidelines for choosing a method are discussed.

Original languageEnglish (US)
Pages (from-to)e185-e191
JournalJournal of Investigative Dermatology
Volume137
Issue number11
DOIs
StatePublished - Nov 2017

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Dermatology
  • Cell Biology

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