Light-sheet autofluorescence lifetime imaging microscopy using a SPAD array detector (Conference Presentation)

Kayvan Samimi; Danielle Desa; Wei Lin; Kurt R. Weiss; Joe Li; Jan Huisken; Jenu V. Chacko; Andreas Velten; Jeremy D. Rogers; Kevin W. Eliceiri; Melissa C. Skala

doi:10.1117/12.2650245

15 March 2023 Light-sheet autofluorescence lifetime imaging microscopy using a SPAD array detector (Conference Presentation)

Kayvan Samimi, Danielle Desa, Wei Lin, Kurt R. Weiss, Joe Li, Jan Huisken, Jenu V. Chacko, Andreas Velten, Jeremy D. Rogers, Kevin W. Eliceiri, Melissa C. Skala

Author Affiliations +

Proceedings Volume PC12386, Single Molecule Spectroscopy and Superresolution Imaging XVI; PC1238605 (2023) https://doi.org/10.1117/12.2650245
Event: SPIE BiOS, 2023, San Francisco, California, United States

Abstract

SPAD array sensors support higher-throughput fluorescence lifetime imaging microscopy (FLIM) by transitioning from laser-scanning to wide-field geometries. While a SPAD camera in epi-fluorescence geometry enables wide-field FLIM of fluorescently labeled samples, label-free imaging of single-cell autofluorescence is not feasible in an epi-fluorescence geometry because background fluorescence from out-of-focus culture medium masks cell autofluorescence and biases lifetime measurements. Here, we address this problem in a proof-of-concept implementation by integrating the SPAD camera in a light-sheet illumination geometry to achieve optical sectioning and limit out-of-focus contributions, enabling label-free wide-field FLIM of single-cell NAD(P)H autofluorescence.

Conference Presentation

Citation Download Citation

Kayvan Samimi, Danielle Desa, Wei Lin, Kurt R. Weiss, Joe Li, Jan Huisken, Jenu V. Chacko, Andreas Velten, Jeremy D. Rogers, Kevin W. Eliceiri, and Melissa C. Skala "Light-sheet autofluorescence lifetime imaging microscopy using a SPAD array detector (Conference Presentation)", Proc. SPIE PC12386, Single Molecule Spectroscopy and Superresolution Imaging XVI, PC1238605 (15 March 2023); https://doi.org/10.1117/12.2650245

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KEYWORDS

Sensors

Detector arrays

Fluorescence lifetime imaging

Microscopy

Cameras

Luminescence

Objectives

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