Fluorescence lifetime: Beating the IRF and interpulse window.
Journal
Biophysical journal
ISSN: 1542-0086
Titre abrégé: Biophys J
Pays: United States
ID NLM: 0370626
Informations de publication
Date de publication:
21 02 2023
21 02 2023
Historique:
received:
09
09
2022
revised:
29
11
2022
accepted:
11
01
2023
pmc-release:
21
02
2024
pubmed:
21
1
2023
medline:
25
2
2023
entrez:
20
1
2023
Statut:
ppublish
Résumé
Fluorescence lifetime imaging captures the spatial distribution of chemical species across cellular environments employing pulsed illumination confocal setups. However, quantitative interpretation of lifetime data continues to face critical challenges. For instance, fluorescent species with known in vitro excited-state lifetimes may split into multiple species with unique lifetimes when introduced into complex living environments. What is more, mixtures of species, which may be both endogenous and introduced into the sample, may exhibit 1) very similar lifetimes as well as 2) wide ranges of lifetimes including lifetimes shorter than the instrumental response function or whose duration may be long enough to be comparable to the interpulse window. By contrast, existing methods of analysis are optimized for well-separated and intermediate lifetimes. Here, we broaden the applicability of fluorescence lifetime analysis by simultaneously treating unknown mixtures of arbitrary lifetimes-outside the intermediate, Goldilocks, zone-for data drawn from a single confocal spot leveraging the tools of Bayesian nonparametrics (BNP). We benchmark our algorithm, termed BNP lifetime analysis, using a range of synthetic and experimental data. Moreover, we show that the BNP lifetime analysis method can distinguish and deduce lifetimes using photon counts as small as 500.
Identifiants
pubmed: 36659850
pii: S0006-3495(23)00030-9
doi: 10.1016/j.bpj.2023.01.014
pmc: PMC9989884
pii:
doi:
Substances chimiques
Coloring Agents
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
672-683Subventions
Organisme : NIGMS NIH HHS
ID : P41 GM103540
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM130745
Pays : United States
Organisme : NIGMS NIH HHS
ID : R01 GM134426
Pays : United States
Informations de copyright
Copyright © 2023 Biophysical Society. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests Authors declare no competing interests.
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