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

TY - JOUR

T1 - Non fitting based FRET-FLIM analysis approaches applied to quantify protein-protein interactions in live cells

AU - Padilla-Parra, Sergi

AU - Auduge, Nicolas

AU - Coppey-Moisan, Maite

AU - Tramier, Marc

PY - 2011/12/1

Y1 - 2011/12/1

N2 - 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.

AB - 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.

KW - Fluorescence lifetime imaging microscopy (FLIM)

KW - Förster resonance energy transfer (FRET)

KW - Nanoscopy

KW - Quantitative fluorescence microscopy

UR - http://www.scopus.com/inward/record.url?scp=84859840792&partnerID=8YFLogxK

U2 - 10.1007/s12551-011-0047-6

DO - 10.1007/s12551-011-0047-6

M3 - Review article

AN - SCOPUS:84859840792

SN - 1867-2450

VL - 3

SP - 63

EP - 70

JO - Biophysical Reviews

JF - Biophysical Reviews

IS - 2

ER -