GFP-based FRET analysis in live cells

Christina L. Takanishi, Ekaterina A. Bykova, Wei Cheng, Jie Zheng

Research output: Contribution to journalArticle

83 Scopus citations

Abstract

Fluorescence resonance energy transfer (FRET) is a widely utilized optical technique for measuring small distances of 1-10 nm in live cells. In recent years, its application has been greatly popularized by the discovery of green fluorescent protein (GFP) and many improved variants which make good donor-acceptor fluorophore pairs. GFP-based proteins are structurally stable, relatively inert, and can be reliably attached to points of interest. The combination of easy access to the GFP-based FRET technique and its obvious usefulness in many applications can lead to complacency. Potential problems such as light contaminants, e.g., bleed-through and cross-talk, and inconsistent donor and acceptor concentrations are easily overlooked and can lead to errors in FRET calculation and data interpretation. In this article, we outline possible pitfalls of GFP-based FRET and approaches that address these issues, including a "Spectra FRET" technique that can be easily applied to live cell studies.

Original languageEnglish (US)
Pages (from-to)132-139
Number of pages8
JournalBrain Research
Volume1091
Issue number1
DOIs
StatePublished - May 26 2006

Keywords

  • Fluorescence resonance energy transfer
  • Green fluorescent protein
  • microscopy
  • Protein-protein interaction
  • Spectra FRET
  • Spectroscopy

ASJC Scopus subject areas

  • Neuroscience(all)
  • Clinical Neurology
  • Developmental Biology
  • Molecular Biology

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