Zickus, VytautasWu, Ming-LoMorimoto, KazuhiroKapitany, ValentinFatima, AreebaTurpin, AlexInsall, RobertWhitelaw, JamieMachesky, LauraBruschini, ClaudioFaccio, DanieleCharbon, Edoardo2021-03-262021-03-262021-03-262020-12-0210.1038/s41598-020-77737-0https://infoscience.epfl.ch/handle/20.500.14299/176569WOS:000600424800005Fluorescence 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.Multidisciplinary SciencesScience & Technology - Other Topicscmostimearraygreenfrettext::journal::journal article::research article