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Conventional plasma membrane probes often suffer from progressive intracellular internalization and nonspecific serum\u2010associated fluorescence. Herein, we report PM580, a serum\u2010resistant fluorogenic probe that enables long\u2010term, wash\u2010free, and surface\u2010confined plasma membrane imaging in live cells. PM580 remains weakly emissive as quenched supramolecular assemblies in aqueous media but becomes strongly fluorescent upon redistribution into lipid bilayers. Structure\u2013property analysis and molecular modeling suggest that the carboxylic acid\u2010containing acceptor may serve as an interfacial positioning element that favors a membrane\u2010inserted orientation, whereas excessive alkyl\u2010chain extension promotes intracellular accumulation and compromises surface selectivity. This balanced architecture enables strong plasma membrane activation with minimal responses to serum albumin or mucin, allowing direct imaging in complete medium and mucin\u2010rich intestinal epithelial monolayers. PM580 further enabled real\u2010time visualization of junctional disruption, membrane tethering, and cell\u2013cell dissociation induced by\n                    <jats:italic>P. gingivalis<\/jats:italic>\n                    conditioned medium. These findings identify a favorable structural balance that supports serum\u2010compatible and long\u2010term plasma membrane imaging.\n                  <\/jats:p>","DOI":"10.1002\/smll.76144","type":"journal-article","created":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T09:43:21Z","timestamp":1791971001000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A Serum\u2010Resistant Plasma Membrane Probe for Real\u2010Time Visualization of Pathogen\u2010Induced Epithelial Barrier Dysfunction"],"prefix":"10.1002","author":[{"given":"Yujeong","family":"Choi","sequence":"first","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"},{"name":"Department of Life Sciences Korea University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4906-7583","authenticated-orcid":false,"given":"Sunhwa","family":"Jung","sequence":"additional","affiliation":[{"name":"Department of Applied Chemistry Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"},{"name":"Center for Bionano Intelligence Education and Research Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong Min","family":"Kang","sequence":"additional","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"},{"name":"Department of Life Sciences Korea University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5262-9746","authenticated-orcid":false,"given":"Lizaveta","family":"Gotina","sequence":"additional","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Minji","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Applied Chemistry Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"},{"name":"Center for Bionano Intelligence Education and Research Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eunmi","family":"Ha","sequence":"additional","affiliation":[{"name":"Department of Applied Chemistry Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3507-4089","authenticated-orcid":false,"given":"Sungsu","family":"Lim","sequence":"additional","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"},{"name":"Division of Bio\u2010Medical Science &amp; Technology KIST School Korea University of Science and Technology (UST)  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ae Nim","family":"Pae","sequence":"additional","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"},{"name":"Division of Bio\u2010Medical Science &amp; Technology KIST School Korea University of Science and Technology (UST)  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jaeson","family":"Jang","sequence":"additional","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"},{"name":"Division of Bio\u2010Medical Science &amp; Technology KIST School Korea University of Science and Technology (UST)  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4570-5130","authenticated-orcid":false,"given":"Evgeni","family":"Ponimaskin","sequence":"additional","affiliation":[{"name":"Department of Cellular Neurophysiology Hannover Medical School  Hannover Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4607-924X","authenticated-orcid":false,"given":"Ja\u2010Hyun","family":"Baik","sequence":"additional","affiliation":[{"name":"Department of Life Sciences Korea University  Seoul Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0867-4416","authenticated-orcid":false,"given":"Sun\u2010Joon","family":"Min","sequence":"additional","affiliation":[{"name":"Department of Applied Chemistry Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"},{"name":"Center for Bionano Intelligence Education and Research Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"},{"name":"Department of Energy and Bio Sciences Hanyang University  Ansan Gyeonggi\u2010do Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8682-120X","authenticated-orcid":false,"given":"Yun Kyung","family":"Kim","sequence":"additional","affiliation":[{"name":"Center for Brain Disorders Brain Science Institute Korea Institute of Science and Technology (KIST)  Seoul Republic of Korea"},{"name":"Division of Bio\u2010Medical Science &amp; 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