{"doi":"10.1016/j.redox.2020.101549","title":"Single cell-based fluorescence lifetime imaging of intracellular oxygenation and metabolism","abstract":"Oxidation-reduction chemistry is fundamental to the metabolism of all living organisms, and hence quantifying the principal redox players is important for a comprehensive understanding of cell metabolism in normal and pathological states. In mammalian cells, this is accomplished by measuring oxygen partial pressure (pO2) in parallel with free and enzyme-bound reduced nicotinamide adenine dinucleotide (phosphate) [H] (NAD(P)H) and flavin adenine dinucleotide (FAD, a proxy for NAD+). Previous optical methods for these measurements had accompanying problems of cytotoxicity, slow speed, population averaging, and inability to measure all redox parameters simultaneously. Herein we present a Förster resonance energy transfer (FRET)-based oxygen sensor, Myoglobin-mCherry, compatible with fluorescence lifetime imaging (FLIM)-based measurement of nicotinamide coenzyme state. This offers a contemporaneous reading of metabolic activity through real-time, non-invasive, cell-by-cell intracellular pO2 and coenzyme status monitoring in living cells. Additionally, this method reveals intracellular spatial heterogeneity and cell-to-cell variation in oxygenation and coenzyme states.","journal":"Redox Biology","year":2020,"id":91385,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":45,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9487,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":458259,"name":"Branden Roarke","orcid":null,"position":1,"is_corresponding":false},{"id":458260,"name":"Greg Alspaugh","orcid":null,"position":2,"is_corresponding":false},{"id":457509,"name":"Anahit Gevorgyan","orcid":"0000-0003-0587-0710","position":3,"is_corresponding":false},{"id":457510,"name":"Alessio Andreoni","orcid":"0000-0003-4415-2770","position":4,"is_corresponding":false},{"id":371372,"name":"Alessandra Pasut","orcid":"0000-0002-0895-4883","position":5,"is_corresponding":false},{"id":401702,"name":"Dan L. Sackett","orcid":"0000-0002-3492-1866","position":6,"is_corresponding":false},{"id":438071,"name":"Jay R. Knutson","orcid":"0000-0001-9990-1738","position":7,"is_corresponding":false},{"id":458258,"name":"Rozhin Penjweini","orcid":null,"position":0,"is_corresponding":true}],"reference_count":45,"raw_metadata":null,"created_at":"2026-07-18T22:28:45.409789Z","pmid":"32403080","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":[]}