Three-color single-molecule maximum likelihood analysis of folding and binding of diffusing molecules.

ABSTRACT: Three-color single-molecule Förster resonance energy transfer (FRET) is a valuable tool to study conformational dynamics of macromolecules. In this work, we present a maximum likelihood method for analyzing three-color fluorescence bursts collected from freely diffusing molecules in confocal microscopy. In three-color single-molecule FRET measurements, the third dye with the longest wavelength typically has a much lower quantum yield than the other two dyes, which leads to significantly reduced brightness, particularly for molecular states with high energy transfer to the third dye. This results in biased detection of bursts and inaccurate estimation of kinetic parameters. We extend the previously developed two-color maximum likelihood method (burstML) to the analysis of three-color data,

SUBMITTER: Feng CJ 

PROVIDER: S-EPMC12424054 | biostudies-literature | 2025 Sep

REPOSITORIES: biostudies-literature