{"doi":"10.1364/boe.568712","title":"High-speed wide-field fluorescence lifetime imaging for intraoperative tumor visualization and in vivo multiplexing","abstract":"We present a technique for fast wide-field fluorescence lifetime (FLT) imaging using simultaneous steady-state and time- (or frequency-) domain measurements acquired in a single shot and exploiting the theoretical dependence of fluorescence intensity on FLT. Using theory, simulations, and in vitro experiments, we show that the single-shot FLT (S-FLT) method can provide absolute FLTs at a higher signal-to-noise ratio compared to other high-speed FLT imaging techniques, without the need for sample-dependent system calibration or model training. We demonstrate that S-FLT can provide accurate tumor delineation in real-time during surgical resections in preclinical tumor models in vivo and in clinical patient specimens ex vivo . We also extend S-FLT for real-time multiplexing and demonstrate this technique for quantifying dynamic FLT changes in vitro and for multiplexing of organ-targeted NIR fluorophores in vivo . Our results suggest that S-FLT can enable real-time quantitative high-throughput studies in preclinical applications and accurate tumor margin delineation in real-time during surgeries.","journal":"Biomedical Optics Express","year":2025,"id":526459,"datarank":0.20794415416798362,"base_score":1.3862943611198906,"endowment":1.3862943611198906,"self_citation_contribution":0.20794415416798362,"citation_network_contribution":0.0,"self_endowment_contribution":0.20794415416798362,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":3,"citer_count":3,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9509,"is_data_producer":true,"deposit_databanks":{"figshare":["10.6084/m9.figshare.29834471"]},"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":873524,"name":"Rahul Pal","orcid":"0000-0003-2062-2678","position":1,"is_corresponding":false},{"id":229907,"name":"Steven S. Hou","orcid":null,"position":2,"is_corresponding":false},{"id":256154,"name":"Hak Soo Choi","orcid":"0000-0002-7982-6483","position":3,"is_corresponding":false},{"id":228741,"name":"Brian J. Bacskai","orcid":"0000-0002-2584-6702","position":4,"is_corresponding":false},{"id":526574,"name":"Alexei Bogdanov","orcid":"0000-0003-4806-5869","position":5,"is_corresponding":false},{"id":50930,"name":"Kenneth K. Tanabe","orcid":"0000-0003-1920-3417","position":6,"is_corresponding":false},{"id":465044,"name":"Mark A. Varvares","orcid":"0000-0002-6570-5672","position":7,"is_corresponding":false},{"id":1401999,"name":"Anand Kumar","orcid":"0000-0001-9868-6220","position":8,"is_corresponding":false},{"id":1012820,"name":"Murali Krishnamoorthy","orcid":null,"position":0,"is_corresponding":true}],"reference_count":60,"raw_metadata":null,"created_at":"2026-07-19T02:50:30.402772Z","pmid":"41368082","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":[]}