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However, flexible organic afterglow molecules with well\u2010characterized structure\u2013luminescence intensity relationships for high\u2010contrast activatable imaging remain scarce. Here, we report for the first time that coumarin derivatives exhibit bright afterglow luminescence and reveal that rigidified coumarin structures display stronger emission than their flexible and backbone analogues. Further, we designed MP molecules bearing boronic ester and methylenecyclobutane moieties and integrated them with coumarin to construct the activatable afterglow probe C545\u2013MP NPs. Oxidation of the boronic ester by ONOO\n                    <jats:sup>\u2212<\/jats:sup>\n                    primes the probe, whereas subsequent irradiation induces coumarin\u2010mediated singlet oxygen generation that reacts with the methylenecyclobutane unit to initiate afterglow emission. By virtue of the elimination of autofluorescence and high activation contrast, C545\u2013MP NPs enable precise assessing therapeutic efficacy of ulcerative colitis and Parkinson\u2032s disease.\n                  <\/jats:p>","DOI":"10.1002\/anie.3724922","type":"journal-article","created":{"date-parts":[[2026,5,4]],"date-time":"2026-05-04T11:40:08Z","timestamp":1777894808000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Engineering Coumarin\u2010Based Afterglow Luminescence Probes for Activatable Imaging Peroxynitrite In Vivo"],"prefix":"10.1002","volume":"65","author":[{"ORCID":"https:\/\/orcid.org\/0009-0003-7249-701X","authenticated-orcid":false,"given":"Youjuan","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory, Zhejiang Cancer Hospital, Zhejiang Provincial Key Laboratory of Protein Detection Technology and Diagnostic Equipment, The Key Laboratory of Zhejiang 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