Development of a high-speed confocal line scanning FLIM microscope for live cell imaging (Conference Presentation)

Simon P. Poland; Hanning Mai; Anneliese Jarman; Conor Treacy; Ahmet Erdogan; Neil Finlayson; Maddy Parsons; Claire Wells; Robert Henderson

doi:10.1117/12.2650206

16 March 2023 Development of a high-speed confocal line scanning FLIM microscope for live cell imaging (Conference Presentation)

Simon P. Poland, Hanning Mai, Anneliese Jarman, Conor Treacy, Ahmet Erdogan, Neil Finlayson, Maddy Parsons, Claire Wells, Robert Henderson

Author Affiliations +

Proceedings Volume PC12390, High-Speed Biomedical Imaging and Spectroscopy VIII; PC123900T (2023) https://doi.org/10.1117/12.2650206
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

We report the development of a novel confocal line-scanning microscope capable of acquiring video-frame rate TCSPC-based FLIM. The system consists of a one-dimensional laser beam, which is optically conjugated to a 1024×16 single photon avalanche diode(SPAD) based line-imaging CMOS(1), with 23.78 μm pixel pitch at 49.31% fill factor. Incorporation of on-chip histogramming on the line-sensor facilitates the acquisition of up to 16.5 Giga-photon counts/s, enabling operation 66 times faster than our previously reported bespoke high speed FLIM platforms. We will demonstrate its use in live-cell imaging investigating the roles that PAK proteins play in regulation of cytoskeletal dynamics.

Conference Presentation

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Simon P. Poland, Hanning Mai, Anneliese Jarman, Conor Treacy, Ahmet Erdogan, Neil Finlayson, Maddy Parsons, Claire Wells, and Robert Henderson "Development of a high-speed confocal line scanning FLIM microscope for live cell imaging (Conference Presentation)", Proc. SPIE PC12390, High-Speed Biomedical Imaging and Spectroscopy VIII, PC123900T (16 March 2023); https://doi.org/10.1117/12.2650206

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KEYWORDS

Fluorescence lifetime imaging

Microscopes

Confocal microscopy

Live cell imaging

Luminescence

In vivo imaging

Surface plasmons

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