Determination of the size of lipid rafts studied through single-molecule FRET simulations

Fluorescence resonance energy transfer (FRET) is a high-resolution technique that allows the characterization of spatial and temporal properties of biological structures and mechanisms. In this work, we developed an in silico single-molecule FRET methodology to study the dynamics of fluorophores ins...

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Bibliographic Details
Published in:Biophysical journal 2021-06, Vol.120 (11), p.2287-2295
Main Authors: Hernández-Adame, Pablo Luis, Meza, Ulises, Rodríguez-Menchaca, Aldo A., Sánchez-Armass, Sergio, Ruiz-García, Jaime, Gomez, Eduardo
Format: Article
Language:English
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Summary:Fluorescence resonance energy transfer (FRET) is a high-resolution technique that allows the characterization of spatial and temporal properties of biological structures and mechanisms. In this work, we developed an in silico single-molecule FRET methodology to study the dynamics of fluorophores inside lipid rafts. We monitored the fluorescence of a single acceptor molecule in the presence of several donor molecules. By looking at the average fluorescence, we selected events with single acceptor and donor molecules, and we used them to determine the raft size in the range of 5–16 nm. We conclude that our method is robust and insensitive to variations in the diffusion coefficient, donor density, or selected fluorescence threshold.
ISSN:0006-3495
1542-0086
DOI:10.1016/j.bpj.2021.04.003