{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T13:06:07Z","timestamp":1790859967853,"version":"4.1.0"},"reference-count":55,"publisher":"Wiley","issue":"12","license":[{"start":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T00:00:00Z","timestamp":1733961600000},"content-version":"vor","delay-in-days":11,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Extracellular Vesicle"],"published-print":{"date-parts":[[2024,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Small extracellular vesicles and nanoparticles (sEVPs) are cell\u2010secreted entities with potential as diagnostic biomarkers and therapeutic vehicles. However, significant intrinsic sEVP heterogeneity impedes analysis and understanding of their composition and functions. We employ multidimensional fluorescent labelling on sEVPs, leveraging the robustness of a newly developed membrane probe\u2014conjugated oligoelectrolytes (COEs), and conduct total internal reflection fluorescence (TIRF) microscopy on sEVP arrays. These arrays comprise single sEVPs anchored to a soft material functionalized surface with little bias. We then develop an enhanced algorithm for colocalization analysis of the multiple labels on individual sEVPs and perform deep profiling of particle content. We categorize sEVPs derived from the same cell type into seven distinct subpopulations\u2014some vesicular whereas others non\u2010vesicular, and we demonstrate that sEVPs from four cell types exhibit quantitatively distinguishable subpopulation distributions. Furthermore, we gain insights into specific particle features within each subpopulation, including CD63 counts, relative particle size, relative concentration of cargoes, and correlations among different cargoes. This high\u2010content analysis reveals common cargo sorting features in sEVP subpopulations across different cell types and suggests new statistics within the sEVP inherent heterogeneity that could differentiate sEVPs from two types of cancer cells and two types of normal cells. Collectively, our study presents a robust single\u2010sEVP characterization platform, combining high\u2010content imaging with comprehensive analysis. This platform is poised to advance sEVP\u2010based theranostic assays and facilitate exploration into disease\u2010associated sEVP biogenesis and sEVP\u2010mediated intercellular communication.<\/jats:p>","DOI":"10.1002\/jev2.12520","type":"journal-article","created":{"date-parts":[[2024,12,12]],"date-time":"2024-12-12T06:37:41Z","timestamp":1733985461000},"source":"Crossref","is-referenced-by-count":10,"title":["Direct imaging with multidimensional labelling and high\u2010content analysis allows quantitative categorization and characterizations of individual small extracellular vesicles and nanoparticles (sEVPs)"],"prefix":"10.1002","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0596-1156","authenticated-orcid":false,"given":"Simou","family":"Sun","sequence":"first","affiliation":[{"name":"Institute for Digital Molecular Analytics and Science Nanyang Technological University  Singapore Singapore"},{"name":"Current address: Department of Chemistry Stony Brook University  New York United\u00a0States"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sarah J.","family":"Cox\u2010V\u00e1zquez","sequence":"additional","affiliation":[{"name":"Department of Chemistry National University of Singapore  Singapore Singapore"},{"name":"Institute for Functional Intelligent Materials National University of Singapore  Singapore Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nam\u2010Joon","family":"Cho","sequence":"additional","affiliation":[{"name":"School of Materials Science and Engineering Nanyang Technological University  Singapore Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guillermo C.","family":"Bazan","sequence":"additional","affiliation":[{"name":"Institute for Digital Molecular Analytics and Science Nanyang Technological University  Singapore Singapore"},{"name":"Department of Chemistry National University of Singapore  Singapore Singapore"},{"name":"Institute for Functional Intelligent Materials National University of Singapore  Singapore Singapore"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jay T.","family":"Groves","sequence":"additional","affiliation":[{"name":"Institute for Digital Molecular Analytics and Science Nanyang Technological University  Singapore Singapore"},{"name":"Department of Chemistry University of California  Berkeley California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2024,12,12]]},"reference":[{"key":"e_1_2_8_2_1","unstructured":"https:\/\/clinicaltrials.gov\/"},{"key":"e_1_2_8_3_1","unstructured":"https:\/\/github.com\/UU\u2010cellbiology\/ComDet"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/smll.202008155"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1080\/20013078.2019.1663043"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep37246"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep37246"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.3791\/62988"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2208993119"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abm3453"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms221910510"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1002\/jev2.12047"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1186\/s12943-022-01671-0"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41565-021-00931-2"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.14107"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.02.029"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tcb.2023.01.002"},{"key":"e_1_2_8_18_1","first-page":"13470","volume-title":"Angewandte Chemie International Edition","author":"Jiang Y.","year":"2021"},{"issue":"131","key":"e_1_2_8_19_1","article-title":"Sample preparation and imaging of exosomes by transmission electron microscopy","volume":"00","author":"Jung M. 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