Fluorescence lifetime imaging with a megapixel SPAD camera and neural network lifetime estimation.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
02 12 2020
02 12 2020
Historique:
received:
29
08
2020
accepted:
06
11
2020
entrez:
3
12
2020
pubmed:
4
12
2020
medline:
4
12
2020
Statut:
epublish
Résumé
Fluorescence lifetime imaging microscopy (FLIM) is a key technology that provides direct insight into cell metabolism, cell dynamics and protein activity. However, determining the lifetimes of different fluorescent proteins requires the detection of a relatively large number of photons, hence slowing down total acquisition times. Moreover, there are many cases, for example in studies of cell collectives, where wide-field imaging is desired. We report scan-less wide-field FLIM based on a 0.5 MP resolution, time-gated Single Photon Avalanche Diode (SPAD) camera, with acquisition rates up to 1 Hz. Fluorescence lifetime estimation is performed via a pre-trained artificial neural network with 1000-fold improvement in processing times compared to standard least squares fitting techniques. We utilised our system to image HT1080-human fibrosarcoma cell line as well as Convallaria. The results show promise for real-time FLIM and a viable route towards multi-megapixel fluorescence lifetime images, with a proof-of-principle mosaic image shown with 3.6 MP.
Identifiants
pubmed: 33268900
doi: 10.1038/s41598-020-77737-0
pii: 10.1038/s41598-020-77737-0
pmc: PMC7710711
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
20986Subventions
Organisme : Swiss National Science Foundation
ID : CRSII5_177165
Pays : Switzerland
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