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Fluorescence lifetime imaging with a megapixel SPAD camera and neural network lifetime estimation.

Abstract：

: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.

journal_name

Sci Rep

journal_title

Scientific reports

authors

Zickus V,Wu ML,Morimoto K,Kapitany V,Fatima A,Turpin A,Insall R,Whitelaw J,Machesky L,Bruschini C,Faccio D,Charbon E

doi

10.1038/s41598-020-77737-0

subject

Has Abstract

pub_date

2020-12-02 00:00:00

pages

20986

issue

1

issn

2045-2322

pii

10.1038/s41598-020-77737-0

journal_volume

10

pub_type

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