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

Vytautas Zickus, Ming-Lo Wu, Kazuhiro Morimoto, Valentin Kapitany, Areeba Fatima, Alex Turpin, Robert Insall, Jamie Whitelaw, Larua Machesky, Claudio Bruschini, Daniele Faccio*, Edoardo Charbon*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

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.
Original languageEnglish
Article number20986
Number of pages10
JournalScientific Reports
Volume10
DOIs
Publication statusPublished - 2 Dec 2020
Externally publishedYes

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