Zickus, V. et al. (2020) Fluorescence lifetime imaging with a megapixel SPAD camera and neural network lifetime estimation. Scientific Reports, 10, 20986. (doi: 10.1038/s41598-020-77737-0) (PMID:33268900) (PMCID:PMC7710711)
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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.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Machesky, Professor Laura and Whitelaw, Mr Jamie and Turpin, Dr Alejandro and Zickus, Dr Vytautas and Kapitany, Mr Valentin and Insall, Professor Robert and Fatima, Dr Areeba and Faccio, Professor Daniele |
| Authors: | Zickus, V., Wu, M.-L., Morimoto, K., Kapitany, V., Fatima, A., Turpin, A., Insall, R., Whitelaw, J., Machesky, L., Bruschini, C., Faccio, D., and Charbon, E. |
| College/School: | College of Medical Veterinary and Life Sciences > School of Cancer Sciences College of Science and Engineering > School of Computing Science College of Science and Engineering > School of Physics and Astronomy |
| Journal Name: | Scientific Reports |
| Publisher: | Nature Research |
| ISSN: | 2045-2322 |
| ISSN (Online): | 2045-2322 |
| Copyright Holders: | Copyright © 2020 The Authors |
| First Published: | First published in Scientific Reports 10: 20986 |
| Publisher Policy: | Reproduced under a Creative Commons License |
| Data DOI: | 10.5525/gla.researchdata.1012 |
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