Reducing the Detrimental Effects of Saturation Phenomena in FRET Microscopy

Tímea Szendi-Szatmári, Ágnes Szabó, János Szöllösi, Peter Nagy

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Although Förster resonance energy transfer (FRET) is one of the most widely used biophysical methods in biology, the effect of high excitation intensity, leading to donor and acceptor saturation, has not been addressed previously. Here, we present a formalism for the experimental determination of the FRET efficiency at high excitation intensity when saturation of both the donor and the acceptor significantly affect conventional FRET calculations. We show that the proposed methodology significantly reduces the dependence of the FRET efficiency on excitation intensity, which otherwise significantly distorts FRET calculations at high excitation intensities commonly used in experiments. The work presented here adds additional rigor to the FRET-based investigation of protein interactions and strengthens the device independence of such results.

Original languageEnglish
Pages (from-to)6378-6382
Number of pages5
JournalAnalytical Chemistry
Volume91
Issue number9
DOIs
Publication statusPublished - May 7 2019

ASJC Scopus subject areas

  • Analytical Chemistry

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