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research article

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

Zickus, Vytautas
•
Wu, Ming-Lo  
•
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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s41598-020-77737-0.pdf

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Publisher's Version

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http://purl.org/coar/version/c_970fb48d4fbd8a85

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openaccess

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CC BY

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1.69 MB

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Adobe PDF

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6040c8714917300a873d8a2535a77776

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