{"doi":"10.3389/fbioe.2025.1557021","title":"Autofluorescence lifetime imaging classifies human B and NK cell activation state","abstract":"New non-destructive tools with single-cell resolution are needed to reliably assess B cell and NK cell function for applications including adoptive cell therapy and immune profiling. Optical metabolic imaging (OMI) is a label-free method that measures the autofluorescence intensity and lifetime of the metabolic cofactors NAD(P)H and FAD to quantify metabolism at a single-cell level. Here, we demonstrate that OMI can resolve metabolic changes between primary human quiescent and IL-4/anti-CD40 activated B cells and between quiescent and IL-12/IL-15/IL-18 activated NK cells. We found that stimulated B and NK cells had an increased proportion of free compared to protein-bound NAD(P)H, a reduced redox state, and produced more lactate compared to control cells. The NAD(P)H mean fluorescence lifetime decreased in the stimulated B and NK cells compared to control cells. Random forest models classified B cells and NK cells according to activation state (CD69 + ) based on OMI variables with an accuracy of 93%. Our results show that autofluorescence lifetime imaging can accurately assess B and NK cell activation in a label-free, non-destructive manner.","journal":"Frontiers in Bioengineering and Biotechnology","year":2025,"id":525451,"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":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9555,"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":990048,"name":"Jeremiah Riendeau","orcid":"0000-0002-3014-2849","position":1,"is_corresponding":false},{"id":253455,"name":"Kelsey Tweed","orcid":"0000-0002-6772-6784","position":2,"is_corresponding":false},{"id":572714,"name":"Peter Rehani","orcid":"0000-0002-7344-2884","position":3,"is_corresponding":false},{"id":744913,"name":"Kayvan Samimi","orcid":"0000-0002-2566-612X","position":4,"is_corresponding":false},{"id":264988,"name":"Dan L. Pham","orcid":null,"position":5,"is_corresponding":false},{"id":256690,"name":"Isabel Jones","orcid":null,"position":6,"is_corresponding":false},{"id":1088332,"name":"Elizabeth M. Maly","orcid":null,"position":7,"is_corresponding":false},{"id":451648,"name":"Emmanuel Contreras Guzman","orcid":"0000-0002-1929-3961","position":8,"is_corresponding":false},{"id":277445,"name":"Matthew H. Forsberg","orcid":"0000-0002-0514-6354","position":9,"is_corresponding":false},{"id":1163715,"name":"Ankita Shahi","orcid":null,"position":10,"is_corresponding":false},{"id":1400608,"name":"Lucia Hockerman","orcid":null,"position":11,"is_corresponding":false},{"id":282369,"name":"José M. Ayuso","orcid":"0000-0002-9414-1845","position":12,"is_corresponding":false},{"id":277448,"name":"Christian M. Capitini","orcid":"0000-0002-2276-6731","position":13,"is_corresponding":false},{"id":253453,"name":"Alex J. Walsh","orcid":"0000-0003-3832-8207","position":14,"is_corresponding":false},{"id":108082,"name":"Melissa C. Skala","orcid":"0000-0002-6320-7637","position":15,"is_corresponding":false},{"id":726955,"name":"Rebecca Schmitz","orcid":"0000-0001-5266-697X","position":0,"is_corresponding":true}],"reference_count":63,"raw_metadata":{"citation_network_status":"fetched"},"created_at":"2026-07-19T02:50:21.274116Z","pmid":"40256783","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":[]}