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. 2018 Aug;93(1):e58.
doi: 10.1002/cpps.58. Epub 2018 Jul 9.

Imaging Protein-Protein Interactions by Förster Resonance Energy Transfer (FRET) Microscopy in Live Cells

Javier Manzella-Lapeira  1 , Joseph A Brzostowski  1
Affiliations

Imaging Protein-Protein Interactions by Förster Resonance Energy Transfer (FRET) Microscopy in Live Cells

Javier Manzella-Lapeira et al. Curr Protoc Protein Sci. 2018 Aug.

Abstract

This updated unit compares three methods to acquire Förster Resonance Energy Transfer (FRET) data in living cells using a confocal microscope: Acceptor photobleaching, Acceptor-sensitized emission FRET, and Donor fluorescence lifetime imaging. Detailed protocols for live cell husbandry, image acquisition, and data analysis are provided. In addition to providing instructions for manufacturer's analysis tool sets, we provide an easy-to-use, MATLAB-based code to calculate FRET efficiency from data obtained using the Acceptor photobleaching or Acceptor-sensitized emission method, which can be freely downloaded. © 2018 by John Wiley & Sons, Inc.

Keywords: FLIM; FRET; confocal microscopy; live cell imaging.

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