{"doi":"10.1117/1.jbo.28.6.066502","title":"Light-sheet autofluorescence lifetime imaging with a single-photon avalanche diode array","abstract":"SignificanceFluorescence 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.AimWe aim to design, validate, and demonstrate a proof-of-concept light-sheet system for NAD(P)H FLIM.ApproachA 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.ResultsAn 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.ConclusionsFLIM 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.","journal":"Journal of Biomedical Optics","year":2023,"id":330913,"datarank":0.5333022092234121,"base_score":3.5553480614894135,"endowment":3.5553480614894135,"self_citation_contribution":0.5333022092234121,"citation_network_contribution":0.0,"self_endowment_contribution":0.5333022092234121,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":34,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.965,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2023-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":407597,"name":"Danielle E. Desa","orcid":"0000-0001-8975-0526","position":1,"is_corresponding":false},{"id":1055247,"name":"Wei Lin","orcid":"0000-0002-3003-0150","position":2,"is_corresponding":false},{"id":635062,"name":"Kurt Weiss","orcid":"0000-0002-6033-3937","position":3,"is_corresponding":false},{"id":1055725,"name":"Joe Li","orcid":null,"position":4,"is_corresponding":false},{"id":480595,"name":"Jan Huisken","orcid":"0000-0001-7250-3756","position":5,"is_corresponding":false},{"id":504541,"name":"Veronika Miskolci","orcid":"0000-0001-7900-4626","position":6,"is_corresponding":false},{"id":342391,"name":"Anna Huttenlocher","orcid":"0000-0001-7940-6254","position":7,"is_corresponding":false},{"id":333432,"name":"Jenu V. Chacko","orcid":"0000-0002-6676-0358","position":8,"is_corresponding":false},{"id":1055248,"name":"Andreas Velten","orcid":"0000-0001-5591-828X","position":9,"is_corresponding":false},{"id":519680,"name":"Jeremy D. Rogers","orcid":"0000-0003-3049-2803","position":10,"is_corresponding":false},{"id":72584,"name":"Kevin W. Eliceiri","orcid":"0000-0001-8678-670X","position":11,"is_corresponding":false},{"id":108082,"name":"Melissa C. Skala","orcid":"0000-0002-6320-7637","position":12,"is_corresponding":false},{"id":744913,"name":"Kayvan Samimi","orcid":"0000-0002-2566-612X","position":0,"is_corresponding":true}],"reference_count":77,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T01:09:14.691473Z","pmid":"37351197","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}