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

Richard Day, Cynthia F. Booker, Ammasi Periasamy

Research output: Contribution to journalArticle

62 Citations (Scopus)

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 - 2008
Externally publishedYes

Fingerprint

Venus
Energy Transfer
Energy transfer
Microscopy
Microscopic examination
Venus (planet)
energy transfer
microscopy
proteins
Proteins
Fluorescence
Cells
life (durability)
fluorescence
Fluorophores
Photons
excitation
Luminance
brightness
Lasers

Keywords

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

ASJC Scopus subject areas

  • Biomedical Engineering
  • Biomaterials
  • Electronic, Optical and Magnetic Materials
  • Atomic and Molecular Physics, and Optics
  • Radiology Nuclear Medicine and imaging
  • Radiological and Ultrasound Technology
  • Clinical Biochemistry

Cite this

Characterization of an improved donor fluorescent protein for Förster resonance energy transfer microscopy. / Day, Richard; Booker, Cynthia F.; Periasamy, Ammasi.

In: Journal of Biomedical Optics, Vol. 13, No. 3, 031203, 2008.

Research output: Contribution to journalArticle

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