Time-resolved burst variance analysis.
Journal
Biophysical reports
ISSN: 2667-0747
Titre abrégé: Biophys Rep (N Y)
Pays: United States
ID NLM: 9918266001106676
Informations de publication
Date de publication:
13 Sep 2023
13 Sep 2023
Historique:
received:
03
05
2023
accepted:
03
07
2023
medline:
10
8
2023
pubmed:
10
8
2023
entrez:
10
8
2023
Statut:
epublish
Résumé
Quantifying biomolecular dynamics has become a major task of single-molecule fluorescence spectroscopy methods. In single-molecule Förster resonance energy transfer (smFRET), kinetic information is extracted from the stream of photons emitted by attached donor and acceptor fluorophores. Here, we describe a time-resolved version of burst variance analysis that can quantify kinetic rates at microsecond to millisecond timescales in smFRET experiments of diffusing molecules. Bursts are partitioned into segments with a fixed number of photons. The FRET variance is computed from these segments and compared with the variance expected from shot noise. By systematically varying the segment size, dynamics at different timescales can be captured. We provide a theoretical framework to extract kinetic rates from the decay of the FRET variance with increasing segment size. Compared to other methods such as filtered fluorescence correlation spectroscopy, recurrence analysis of single particles, and two-dimensional lifetime correlation spectroscopy, fewer photons are needed to obtain reliable timescale estimates, which reduces the required measurement time.
Identifiants
pubmed: 37559939
doi: 10.1016/j.bpr.2023.100116
pii: S2667-0747(23)00017-4
pmc: PMC10406964
doi:
Types de publication
Journal Article
Langues
eng
Pagination
100116Informations de copyright
© 2023 The Authors.
Déclaration de conflit d'intérêts
The authors declare no competing interest.
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