{"doi":"10.1002/mrd.22501","title":"FRAP, FLIM, and FRET: Detection and analysis of cellular dynamics on a molecular scale using fluorescence microscopy","abstract":"<jats:title>SUMMARY</jats:title><jats:sec><jats:label/><jats:p>The combination of fluorescent‐probe technology plus modern optical microscopes allows investigators to monitor dynamic events in living cells with exquisite temporal and spatial resolution. Fluorescence recovery after photobleaching (FRAP), for example, has long been used to monitor molecular dynamics both within cells and on cellular surfaces. Although bound by the diffraction limit imposed on all optical microscopes, the combination of digital cameras and the application of fluorescence intensity information on large‐pixel arrays have allowed such dynamic information to be monitored and quantified. Fluorescence lifetime imaging microscopy (FLIM), on the other hand, utilizes the information from an ensemble of fluorophores to probe changes in the local environment. Using either fluorescence‐intensity or lifetime approaches, fluorescence resonance energy transfer (FRET) microscopy provides information about molecular interactions, with Ångstrom resolution. In this review, we summarize the theoretical framework underlying these methods and illustrate their utility in addressing important problems in reproductive and developmental systems. <jats:italic>Mol. Reprod. Dev. 82: 587–604, 2015. © 2015 Wiley Periodicals, Inc</jats:italic>.</jats:p></jats:sec>","journal":"Molecular Reproduction and Development","year":2015,"id":671674,"datarank":2.358808896887418,"base_score":4.189654742026425,"endowment":4.189654742026425,"self_citation_contribution":0.6284482113039639,"citation_network_contribution":1.730360685583454,"self_endowment_contribution":0.6284482113039639,"citer_contribution":1.730360685583454,"corpus_percentile":null,"corpus_rank":null,"citation_count":65,"citer_count":65,"citers_with_citation_signal":53,"citers_with_endowment":53,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1049465,"name":"Ching‐Wei Chang","orcid":"0000-0001-9239-4403","position":1,"is_corresponding":false},{"id":1754799,"name":"Mary‐Ann Mycek","orcid":null,"position":2,"is_corresponding":false},{"id":1754800,"name":"Richard A. Cardullo","orcid":null,"position":3,"is_corresponding":false},{"id":1754798,"name":"Carla De Los Santos","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"FRAP, FLIM, and FRET: Detection and analysis of cellular dynamics on a molecular scale using fluorescence microscopy","abstract":"<jats:title>SUMMARY</jats:title><jats:sec><jats:label/><jats:p>The combination of fluorescent‐probe technology plus modern optical microscopes allows investigators to monitor dynamic events in living cells with exquisite temporal and spatial resolution. Fluorescence recovery after photobleaching (FRAP), for example, has long been used to monitor molecular dynamics both within cells and on cellular surfaces. Although bound by the diffraction limit imposed on all optical microscopes, the combination of digital cameras and the application of fluorescence intensity information on large‐pixel arrays have allowed such dynamic information to be monitored and quantified. Fluorescence lifetime imaging microscopy (FLIM), on the other hand, utilizes the information from an ensemble of fluorophores to probe changes in the local environment. Using either fluorescence‐intensity or lifetime approaches, fluorescence resonance energy transfer (FRET) microscopy provides information about molecular interactions, with Ångstrom resolution. In this review, we summarize the theoretical framework underlying these methods and illustrate their utility in addressing important problems in reproductive and developmental systems. <jats:italic>Mol. Reprod. Dev. 82: 587–604, 2015. © 2015 Wiley Periodicals, Inc</jats:italic>.</jats:p></jats:sec>","is_dataset_classified":null,"base_score":4.189654742026425,"endowment":4.189654742026425,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"26010322","pmcid":null,"openalex_id":"https://openalex.org/W1549001647","authors":[],"funders":[{"funder_name":"NIH","grant_id":"R01 HD27244","title":null},{"funder_name":"NIH","grant_id":"R01 DE019431","title":null},{"funder_name":"NICHD NIH HHS","grant_id":"R01-HD-27244","title":null}],"total_grants":3,"fwci":5.7634,"citation_percentile":0.95211824,"influential_citations":0,"citation_trend":[{"year":2015,"count":1},{"year":2016,"count":1},{"year":2017,"count":7},{"year":2018,"count":7},{"year":2019,"count":9},{"year":2020,"count":6},{"year":2021,"count":10},{"year":2022,"count":10},{"year":2023,"count":2},{"year":2024,"count":6},{"year":2025,"count":3},{"year":2026,"count":3}],"oa_status":"green","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://hdl.handle.net/2027.42/112297","host_type":"repository"},{"url":"https://hdl.handle.net/2027.42/112297","host_type":"repository"},{"url":"http://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fmrd.22501","host_type":"publisher"},{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fmrd.22501","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1002/mrd.22501","host_type":"publisher"},{"url":"https://doi.org/10.1002/mrd.22501","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/26010322","host_type":"repository"},{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/4515154","host_type":"repository"}],"fields_of_study":["Advanced Fluorescence Microscopy Techniques","Cellular Mechanics and Interactions","Cell Image Analysis Techniques","Animals","Fluorescence Recovery After Photobleaching","Fluorescence Resonance Energy Transfer","Humans","Image Processing, Computer-Assisted","Microscopy, Fluorescence"],"mesh_terms":["Animals","Humans","Image Processing, Computer-Assisted","Microscopy, Fluorescence","Fluorescence Resonance Energy Transfer","Fluorescence Recovery After Photobleaching"],"keywords":["Förster resonance energy transfer","Photobleaching","Microscopy","Fluorescence recovery after photobleaching","Fluorescence-lifetime imaging microscopy","Fluorescence microscope","Fluorescence","Microscope","Biology","Live cell imaging","Resolution (logic)","Confocal microscopy","Temporal resolution","Fluorescence in the life sciences","Biological system","Biophysics","Optics","Physics","Computer science","Artificial intelligence","Cell biology"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Affordable and clean energy"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-08-16T03:56:16.070542Z","pmid":null,"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":[]}