Dynamic real-time subtraction of stray-light and background for multiphoton imaging.
Journal
Biomedical optics express
ISSN: 2156-7085
Titre abrégé: Biomed Opt Express
Pays: United States
ID NLM: 101540630
Informations de publication
Date de publication:
01 Jan 2021
01 Jan 2021
Historique:
received:
20
07
2020
revised:
13
10
2020
accepted:
17
11
2020
entrez:
4
3
2021
pubmed:
5
3
2021
medline:
5
3
2021
Statut:
epublish
Résumé
We introduce a new approach to reduce uncorrelated background signals from fluorescence imaging data, using real-time subtraction of background light. This approach takes advantage of the short fluorescence lifetime of most popular fluorescent activity reporters, and the low duty-cycle of ultrafast lasers. By synchronizing excitation and recording, laser-induced multiphoton fluorescence can be discriminated from background light levels with each laser pulse. We demonstrate the ability of our method to - in real-time - remove image artifacts that in a conventional imaging setup lead to clipping of the signal. In other words, our method enables imaging under conditions that in a conventional setup would yield corrupted data from which no accurate information can be extracted. This is advantageous in experimental setups requiring additional light sources for applications such as optogenetic stimulation.
Identifiants
pubmed: 33659077
doi: 10.1364/BOE.403255
pii: 403255
pmc: PMC7899518
doi:
Types de publication
Journal Article
Langues
eng
Pagination
288-302Informations de copyright
Published by The Optical Society under the terms of the Creative Commons Attribution 4.0 License. Further distribution of this work must maintain attribution to the author(s) and the published article’s title, journal citation, and DOI.
Déclaration de conflit d'intérêts
The authors declare no conflicts of interest.
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