{"doi":"10.1117/1.bios.2.3.030901","title":"High-speed fluorescence lifetime imaging microscopy: techniques, applications, and prospects","abstract":"Significance: Fluorescence lifetime imaging microscopy (FLIM) has been gaining increasing attention due to its capability to provide robust, accurate, and quantitative measurements compared with conventional intensity-based fluorescence microscopy. However, the slow imaging speed of FLIM has long been a major limitation for expanding its applications in biology and medicine. Aim: We aim to discuss recent advancements in FLIM to enhance imaging speed and explore its biomedical applications and future prospects. Approach: We discuss high-speed FLIM techniques by categorizing them into time-domain and frequency-domain approaches, as well as wide-field and beam-scanning imaging schemes, with a focus on their combinations. Results: Recent advances in high-speed FLIM have been primarily driven by innovations in fluorescence detection schemes for wide-field imaging, as well as fluorescence excitation strategies for beam-scanning imaging. By enhancing FLIM imaging speed to levels comparable with intensity-based fluorescence microscopy, the observation of fast cellular dynamics, such as neural spiking, and large-scale image-based analysis of heterogeneous cells have become feasible. In addition, promising directions for the future development of high-speed FLIM include further utilization of fit-free analysis, implementation of video-rate 3D FLIM, and realization of FLIM-based cell sorting. Conclusions: The development of high-speed FLIM promises new opportunities in cell biology, biophysics, and neurology.","journal":"Biophotonics discovery.","year":2025,"id":529799,"datarank":0.3453877639491069,"base_score":2.302585092994046,"endowment":2.302585092994046,"self_citation_contribution":0.3453877639491069,"citation_network_contribution":0.0,"self_endowment_contribution":0.3453877639491069,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":9,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9561,"is_data_producer":false,"deposit_databanks":null,"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":785569,"name":"Fan Li","orcid":"0000-0002-8024-8326","position":1,"is_corresponding":false},{"id":1409163,"name":"Jongchan Park","orcid":"0000-0001-9808-6823","position":2,"is_corresponding":false},{"id":1355966,"name":"Hidenori Endo","orcid":"0000-0003-1472-6032","position":3,"is_corresponding":false},{"id":1409164,"name":"Kuniyasu Niizuma","orcid":"0000-0001-9282-6499","position":4,"is_corresponding":false},{"id":628053,"name":"Liang Gao","orcid":"0000-0001-9983-0626","position":5,"is_corresponding":false},{"id":642721,"name":"Keisuke Goda","orcid":"0000-0001-6302-6038","position":6,"is_corresponding":false},{"id":642708,"name":"Hiroshi Kanno","orcid":"0000-0003-3340-9462","position":0,"is_corresponding":true}],"reference_count":131,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:51:01.235017Z","pmid":"42028255","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":[]}