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Curr Protoc Protein Sci. 2009 Apr;Chapter 19:Unit19.5. doi: 10.1002/0471140864.ps1905s56.

Imaging protein-protein interactions by Förster resonance energy transfer (FRET) microscopy in live cells.

Author information

1
NIH/NIAID-LIG Imaging Facility, Rockville, Maryland, USA.

Abstract

This unit describes an acceptor-sensitized emission FRET method using a confocal microscope for image acquisition. In contrast to acceptor photobleaching, which is an end-point assay that destroys the acceptor fluorophore, the sensitized emission method is amenable for FRET measurements in live cells and can be used to measure changes in FRET efficiency over time. The purpose of this unit is to provide a basic starting point for understanding and performing the sensitized emission method with a simple teaching tool for live-cell imaging. References that discuss the vagaries of acquiring and analyzing FRET between individually tagged molecules are provided.

PMID:
19365789
PMCID:
PMC3568943
DOI:
10.1002/0471140864.ps1905s56
[Indexed for MEDLINE]
Free PMC Article

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