{"doi":"10.1016/j.crmeth.2022.100267","title":"High-resolution imaging of protein secretion at the single-cell level using plasmon-enhanced FluoroDOT assay","abstract":"Secreted proteins mediate essential physiological processes. With conventional assays, it is challenging to map the spatial distribution of proteins secreted by single cells, to study cell-to-cell heterogeneity in secretion, or to detect proteins of low abundance or incipient secretion. Here, we introduce the “FluoroDOT assay,” which uses an ultrabright nanoparticle plasmonic-fluor that enables high-resolution imaging of protein secretion. We find that plasmonic-fluors are 16,000-fold brighter, with nearly 30-fold higher signal-to-noise compared with conventional fluorescence labels. We demonstrate high-resolution imaging of different secreted cytokines in the single-plexed and spectrally multiplexed FluoroDOT assay that revealed cellular heterogeneity in secretion of multiple proteins simultaneously. Using diverse biochemical stimuli, including Mycobacterium tuberculosis infection, and a variety of immune cells such as macrophages, dendritic cells (DCs), and DC-T cell co-culture, we demonstrate that the assay is versatile, facile, and widely adaptable for enhancing biological understanding of spatial and temporal dynamics of single-cell secretome.","journal":"Cell Reports Methods","year":2022,"id":269328,"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":16,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9537,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":932102,"name":"Ekansh Mittal","orcid":"0000-0001-9034-033X","position":1,"is_corresponding":false},{"id":245681,"name":"Jingyi Luan","orcid":"0000-0001-5129-9506","position":2,"is_corresponding":false},{"id":708290,"name":"Samhitha Kolla","orcid":null,"position":3,"is_corresponding":false},{"id":233399,"name":"Monty B. Mazer","orcid":"0000-0002-4159-8585","position":4,"is_corresponding":false},{"id":878530,"name":"Hemant Joshi","orcid":"0000-0001-7151-359X","position":5,"is_corresponding":false},{"id":245683,"name":"Rohit Gupta","orcid":"0000-0001-7570-7527","position":6,"is_corresponding":false},{"id":248393,"name":"Priya Rathi","orcid":null,"position":7,"is_corresponding":false},{"id":245684,"name":"Zheyu Wang","orcid":"0000-0002-8703-6334","position":8,"is_corresponding":false},{"id":245689,"name":"Jeremiah J. Morrissey","orcid":"0000-0002-9911-4811","position":9,"is_corresponding":false},{"id":8685,"name":"J. Ernst","orcid":"0000-0001-9951-6207","position":10,"is_corresponding":false},{"id":932486,"name":"Cynthia Portal‐Celhay","orcid":null,"position":11,"is_corresponding":false},{"id":397524,"name":"Sharon Celeste Morley","orcid":"0000-0001-8424-0121","position":12,"is_corresponding":false},{"id":278133,"name":"Jennifer A. Philips","orcid":"0000-0002-9476-0240","position":13,"is_corresponding":false},{"id":245690,"name":"Srikanth Singamaneni","orcid":"0000-0002-7203-2613","position":14,"is_corresponding":false},{"id":245682,"name":"Anushree Seth","orcid":"0000-0003-2995-2200","position":0,"is_corresponding":true}],"reference_count":58,"raw_metadata":null,"created_at":"2026-07-19T00:27:18.142851Z","pmid":"36046626","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":[]}