Light-sheet autofluorescence lifetime imaging with a single-photon avalanche diode array.

Autor:	Samimi K; Morgridge Institute for Research, Madison, Wisconsin, United States., Desa DE; Morgridge Institute for Research, Madison, Wisconsin, United States., Lin W; University of Wisconsin, Department of Electrical and Computer Engineering, Madison, Wisconsin, United States., Weiss K; Morgridge Institute for Research, Madison, Wisconsin, United States.; University of Wisconsin, Department of Biochemistry, Madison, Wisconsin, United States., Li J; Morgridge Institute for Research, Madison, Wisconsin, United States., Huisken J; Morgridge Institute for Research, Madison, Wisconsin, United States.; Georg-August-University Göttingen, Department of Biology and Psychology, Göttingen, Germany., Miskolci V; University of Wisconsin, Department of Medical Microbiology and Immunology, Madison, Wisconsin, United States.; Rutgers New Jersey Medical School, Center for Cell Signaling, Newark, New Jersey, United States.; Rutgers New Jersey Medical School, Department of Microbiology, Biochemistry and Molecular Genetics, Newark, New Jersey, United States., Huttenlocher A; University of Wisconsin, Department of Medical Microbiology and Immunology, Madison, Wisconsin, United States.; University of Wisconsin, Department of Pediatrics, Madison, Wisconsin, United States., Chacko JV; University of Wisconsin, Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin, United States., Velten A; Morgridge Institute for Research, Madison, Wisconsin, United States.; University of Wisconsin, Department of Electrical and Computer Engineering, Madison, Wisconsin, United States.; University of Wisconsin, Department of Biostatistics and Medical Informatics, Madison, Wisconsin, United States.; University of Wisconsin, McPherson Eye Research Institute, Madison, Wisconsin, United States., Rogers JD; Morgridge Institute for Research, Madison, Wisconsin, United States.; University of Wisconsin, McPherson Eye Research Institute, Madison, Wisconsin, United States.; University of Wisconsin, Department of Ophthalmology and Visual Sciences, Madison, Wisconsin, United States., Eliceiri KW; Morgridge Institute for Research, Madison, Wisconsin, United States.; University of Wisconsin, Laboratory for Optical and Computational Instrumentation, Madison, Wisconsin, United States.; University of Wisconsin, Department of Biostatistics and Medical Informatics, Madison, Wisconsin, United States.; University of Wisconsin, McPherson Eye Research Institute, Madison, Wisconsin, United States.; University of Wisconsin, Department of Biomedical Engineering, Madison, Wisconsin, United States., Skala MC; Morgridge Institute for Research, Madison, Wisconsin, United States.; University of Wisconsin, McPherson Eye Research Institute, Madison, Wisconsin, United States.; University of Wisconsin, Department of Biomedical Engineering, Madison, Wisconsin, United States.
Jazyk:	angličtina
Zdroj:	Journal of biomedical optics [J Biomed Opt] 2023 Jun; Vol. 28 (6), pp. 066502. Date of Electronic Publication: 2023 Jun 21.
DOI:	10.1117/1.JBO.28.6.066502
Abstrakt:	Significance: Fluorescence lifetime imaging microscopy (FLIM) of the metabolic co-enzyme nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] is a popular method to monitor single-cell metabolism within unperturbed, living 3D systems. However, FLIM of NAD(P)H has not been performed in a light-sheet geometry, which is advantageous for rapid imaging of cells within live 3D samples. Aim: We aim to design, validate, and demonstrate a proof-of-concept light-sheet system for NAD(P)H FLIM. Approach: A single-photon avalanche diode camera was integrated into a light-sheet microscope to achieve optical sectioning and limit out-of-focus contributions for NAD(P)H FLIM of single cells. Results: An NAD(P)H light-sheet FLIM system was built and validated with fluorescence lifetime standards and with time-course imaging of metabolic perturbations in pancreas cancer cells with 10 s integration times. NAD(P)H light-sheet FLIM in vivo was demonstrated with live neutrophil imaging in a larval zebrafish tail wound also with 10 s integration times. Finally, the theoretical and practical imaging speeds for NAD(P)H FLIM were compared across laser scanning and light-sheet geometries, indicating a 30 × to 6 × acquisition speed advantage for the light sheet compared to the laser scanning geometry. Conclusions: FLIM of NAD(P)H is feasible in a light-sheet geometry and is attractive for 3D live cell imaging applications, such as monitoring immune cell metabolism and migration within an organism. (© 2023 The Authors.)
Databáze:	MEDLINE
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