Extra kinetic dimensions for label discrimination.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
18 03 2022
18 03 2022
Historique:
received:
04
06
2021
accepted:
23
02
2022
entrez:
19
3
2022
pubmed:
20
3
2022
medline:
6
4
2022
Statut:
epublish
Résumé
Due to its sensitivity and versatility, fluorescence is widely used to detect specifically labeled biomolecules. However, fluorescence is currently limited by label discrimination, which suffers from the broad full width of the absorption/emission bands and the narrow lifetime distribution of the bright fluorophores. We overcome this limitation by introducing extra kinetic dimensions through illuminations of reversibly photoswitchable fluorophores (RSFs) at different light intensities. In this expanded space, each RSF is characterized by a chromatic aberration-free kinetic fingerprint of photochemical reactivity, which can be recovered with limited hardware, excellent photon budget, and minimal data processing. This fingerprint was used to identify and discriminate up to 20 among 22 spectrally similar reversibly photoswitchable fluorescent proteins (RSFPs) in less than 1s. This strategy opens promising perspectives for expanding the multiplexing capabilities of fluorescence imaging.
Identifiants
pubmed: 35304491
doi: 10.1038/s41467-022-29172-0
pii: 10.1038/s41467-022-29172-0
pmc: PMC8933551
doi:
Substances chimiques
Fluorescent Dyes
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1482Informations de copyright
© 2022. The Author(s).
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