{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:19:31Z","timestamp":1792790371972,"version":"4.5.0"},"posted":{"date-parts":[[2026,9,29]]},"group-title":"Bioengineering","reference-count":38,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T00:00:00Z","timestamp":1790640000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"name":"NIAID","id":[{"id":"https:\/\/ror.org\/043z4tv69","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,29]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>Cell membrane-coated nanoparticles (CNPs) are a potentially transformative biomimetic targeted-delivery platform, which can engage therapeutic targets through source-cell membrane protein functions or homotypic interactions. However, their therapeutic development is limited by the current inability to accurately resolve membrane coating completeness, and CNP particle-to-particle variation. Here, we introduce a STochastic Optical Reconstruction Microscopy (STORM)-based approach that resolves population composition and quantifies membrane coating coverage at the single-particle level. Applying it to dendritic cell membrane- and HeLa cell membrane-coated nanoparticles (DCmPs and HeLamPs), we revealed substantial heterogeneity within both particle types and a cell-type dependence of coating outcomes, corroborated by confocal imaging and flow cytometry. We also quantified membrane protein composition, including peptide major histocompatibility complex class I (pMHC-I) among DCmPs, and confirmed that DCmPs preferentially engaged antigen-specific T cells in coculture. This single-particle framework establishes a quantitative standard for CNP characterization, providing a foundation for quality control and rational design.<\/jats:p>\n                <jats:sec>\n                  <jats:title>TABLE OF CONTENT<\/jats:title>\n                  <jats:p>\n                    <jats:fig id=\"ufig1\" position=\"float\" orientation=\"portrait\" fig-type=\"figure\">\n                      <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"755175v1_ufig1\" position=\"float\" orientation=\"portrait\"\/>\n                    <\/jats:fig>\n                  <\/jats:p>\n                  <jats:p>A single-particle super-resolution imaging method is developed to determine the coating composition and completeness across membrane-coated nanoparticles. Examining two types of coated nanoparticles reveals striking variations in coating coverage and protein composition, and shows cell type-dependent coating quality. This technology provides a new detailed and precise quantality control for membrane-coated nanoparticle design.<\/jats:p>\n                <\/jats:sec>","DOI":"10.64898\/2026.09.28.755175","type":"posted-content","created":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T15:55:16Z","timestamp":1790697316000},"source":"Crossref","is-referenced-by-count":0,"title":["Single-Particle STORM Imaging Quantifies Coating Heterogeneity Among Cell Membrane-Coated Nanoparticles"],"prefix":"10.64898","author":[{"given":"Jimmy","family":"Blauser-Wilson","sequence":"first","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ali","family":"Zareein","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sao","family":"Puth","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"},{"name":"The Global Health Research Center, University of Health Sciences, Khan Daun Penh, Phnom Penh, 12000, Cambodia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shruti Sunil","family":"Jadhav","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raymond","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abigail M","family":"Hernandez","sequence":"additional","affiliation":[{"name":"College of Science, University of Texas at San Antonio, San Antonio, TX, 78249, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrea","family":"Joseph","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jialiu","family":"Zeng","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0444-1017","authenticated-orcid":false,"given":"James L","family":"Hougland","sequence":"additional","affiliation":[{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"},{"name":"Department of Chemistry, Syracuse University, Syracuse, NY, 13244, USA"},{"name":"Department of Biology, Syracuse University, Syracuse, NY, 13244, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0570-2493","authenticated-orcid":false,"given":"Yaoying","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Biomedical and Chemical Engineering, Syracuse University, Syracuse, NY 13210, USA,"},{"name":"The BioInspired Institute for Material and Living Systems, Syracuse University, Syracuse, NY 13210, USA;"},{"name":"Department of Microbiology & Immunology, SUNY Upstate Medical University, Syracuse, NY 13210, 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