Fluorescent proteins for FRET microscopy: Monitoring protein interactions in living cells

Richard Day, Michael W. Davidson

Research output: Contribution to journalArticle

61 Citations (Scopus)

Abstract

The discovery and engineering of novel fluorescent proteins (FPs) from diverse organisms is yielding fluorophores with exceptional characteristics for live-cell imaging. In particular, the development of FPs for fluorescence (or Förster) resonance energy transfer (FRET) microscopy is providing important tools for monitoring dynamic protein interactions inside living cells. The increased interest in FRET microscopy has driven the development of many different methods to measure FRET. However, the interpretation of FRET measurements is complicated by several factors including the high fluorescence background, the potential for photoconversion artifacts and the relatively low dynamic range afforded by this technique. Here, we describe the advantages and disadvantages of four methods commonly used in FRET microscopy. We then discuss the selection of FPs for the different FRET methods, identifying the most useful FP candidates for FRET microscopy. The recent success in expanding the FP color palette offers the opportunity to explore new FRET pairs.

Original languageEnglish
Pages (from-to)341-350
Number of pages10
JournalBioEssays
Volume34
Issue number5
DOIs
StatePublished - May 2012

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Energy Transfer
Energy transfer
Microscopy
Microscopic examination
Cells
Monitoring
Proteins
Fluorescence
Fluorophores
Artifacts
Color
Imaging techniques

Keywords

  • Acceptor photobleaching
  • Fluorescence lifetime imaging microscopy (FLIM)
  • Fluorescence resonance energy transfer (FRET)
  • Fluorescent protein
  • Spectral imaging

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Fluorescent proteins for FRET microscopy : Monitoring protein interactions in living cells. / Day, Richard; Davidson, Michael W.

In: BioEssays, Vol. 34, No. 5, 05.2012, p. 341-350.

Research output: Contribution to journalArticle

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