Non fitting based FRET-FLIM analysis approaches applied to quantify protein-protein interactions in live cells

Sergi Padilla-Parra*, Nicolas Auduge, Maite Coppey-Moisan, Marc Tramier

*Corresponding author for this work

Research output: Contribution to journalReview articlepeer-review

23 Citations (Scopus)

Abstract

New imaging methodologies in quantitative fluorescence microscopy and nanoscopy have been developed in the last few years and are beginning to be extensively applied to biological problems, such as the localization and quantification of protein interactions. Fluorescence resonance energy transfer (FRET) detected by fluorescence lifetime imaging microscopy (FLIM) is currently employed not only in biophysics or chemistry but also in bio-medicine, thanks to new advancements in technology and also new developments in data treatment. FRET-FLIM can be a very useful tool to ascertain protein interactions occurring in single living cells. In this review, we stress the importance of increasing the acquisition speed when working in vivo employing Time-Domain FLIM. The development of the new mathematical-based non-fitting methods allows the determining of the fraction of interacting donor without the requirement of high count statistics, and thus allows the performing of high speed acquisitions in FRET-FLIM to still be quantitative.

Original languageEnglish
Pages (from-to)63-70
Number of pages8
JournalBiophysical Reviews
Volume3
Issue number2
DOIs
Publication statusPublished - 1 Dec 2011

Keywords

  • Fluorescence lifetime imaging microscopy (FLIM)
  • Förster resonance energy transfer (FRET)
  • Nanoscopy
  • Quantitative fluorescence microscopy

Fingerprint

Dive into the research topics of 'Non fitting based FRET-FLIM analysis approaches applied to quantify protein-protein interactions in live cells'. Together they form a unique fingerprint.

Cite this