Characterization of an improved donor fluorescent protein for Förster resonance energy transfer microscopy

Richard N. Day, Cynthia F. Booker, Ammasi Periasamy

Research output: Contribution to journalArticle

64 Scopus citations

Abstract

The genetically encoded fluorescent proteins (FP), used in combination with Förster resonance energy transfer (FRET) microscopy, provide the tools necessary for the direct visualization of protein interactions inside living cells. Typically, the Cerulean and Venus variants of the cyan and yellow FPs are used for FRET studies, but there are limitations to their use. Here, Cerulean and the newly developed monomeric Teal FP (mTFP) are compared as FRET donors for Venus using spectral and fluorescence lifetime measurements from living cells. The results demonstrate that when compared to Cerulean, mTFP has increased brightness, optimal excitation using the standard 458-nm laser line, increased photostability, and improved spectral overlap with Venus. In addition, the two-photon excitation and fluorescence lifetime characteristics are determined for mTFP. Together, these measurements indicate that mTFP is an excellent donor fluorophore for FRET studies, and that its use may improve the detection of interactions involving proteins that are difficult to express, or that need to be produced at low levels in cells.

Original languageEnglish (US)
Article number031203
JournalJournal of Biomedical Optics
Volume13
Issue number3
DOIs
StatePublished - Dec 1 2008
Externally publishedYes

Keywords

  • Cerulean
  • Förster resonance energy transfer microscopy
  • Teal fluorescent protein
  • Venus
  • fluorescent proteins
  • live cell imaging
  • protein-protein interactions

ASJC Scopus subject areas

  • Electronic, Optical and Magnetic Materials
  • Biomaterials
  • Atomic and Molecular Physics, and Optics
  • Biomedical Engineering

Fingerprint Dive into the research topics of 'Characterization of an improved donor fluorescent protein for Förster resonance energy transfer microscopy'. Together they form a unique fingerprint.

  • Cite this