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research article

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

Zickus, Vytautas
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Wu, Ming-Lo  
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Morimoto, Kazuhiro  
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December 2, 2020
Scientific Reports

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.

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Type
research article
DOI
10.1038/s41598-020-77737-0
Web of Science ID

WOS:000600424800005

Author(s)
Zickus, Vytautas
Wu, Ming-Lo  
Morimoto, Kazuhiro  
Kapitany, Valentin
Fatima, Areeba
Turpin, Alex
Insall, Robert
Whitelaw, Jamie
Machesky, Laura
Bruschini, Claudio  
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Date Issued

2020-12-02

Publisher

NATURE RESEARCH

Published in
Scientific Reports
Volume

10

Issue

1

Article Number

20986

Subjects

Multidisciplinary Sciences

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Science & Technology - Other Topics

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cmos

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time

•

array

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green

•

fret

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
AQUA  
Available on Infoscience
March 26, 2021
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/176569
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