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We prospectively analyzed 69 surgically obtained tumor specimens from 61 patients with WHO grade 4 gliomas, WHO grade 2\/3 gliomas, meningiomas, and brain metastases before and after ex vivo incubation in 4 \u00b5M water-soluble high-load hypericin-polyvinylpyrrolidone complex (HHL-PVP). Fluorescence lifetime and intensity were quantified using a dual-tap CMOS camera system with a hypericin-specific 575\u2013615\u00a0nm bandpass filter. HHL-PVP incubation significantly increased both fluorescence intensity and lifetime in all tumor entities, including specimens without visible 5-ALA fluorescence. An independently derived combined lifetime\/intensity regression model discriminated pre- from post-incubation measurements with an area under the receiver operating characteristic curve of 0.975, sensitivity of 98.6%, and specificity of 82.6%; entity-specific areas under the curve ranged from 0.933 to 1.000. These findings support robust hypericin-associated signal detection across major brain tumor entities after ex vivo incubation and provide a rationale for future in vivo evaluation of HHL-PVP in fluorescence-guided neuro-oncological surgery.<\/jats:p>","DOI":"10.1038\/s41598-026-62969-3","type":"journal-article","created":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T17:04:31Z","timestamp":1784567071000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Fluorescence lifetime and intensity detection of water-soluble hypericin accumulation in ex-vivo glioma, meningioma, and brain metastasis tissue"],"prefix":"10.1038","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-5478-0792","authenticated-orcid":false,"given":"Mario","family":"Mischkulnig","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Reichert","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lionel","family":"Wightman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vanessa","family":"Roth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marijke","family":"Hoelz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandra","family":"Klang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lisa I.","family":"K\u00f6rner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Barbara","family":"Kiesel","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Djenana","family":"Vejzovic","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mikael T.","family":"Erkkilae","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angelika","family":"Unterhuber","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rainer","family":"Leitgeb","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Beate","family":"Rinner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Kubin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marco","family":"Andreana","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Georg","family":"Widhalm","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2026,7,20]]},"reference":[{"issue":"8","key":"62969_CR1","doi-asserted-by":"publisher","first-page":"1231","DOI":"10.1093\/neuonc\/noab106","volume":"23","author":"DN Louis","year":"2021","unstructured":"Louis, D. 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MM and GW received research funding from Hypericum Life Sciences GmbH. The remaining authors declare no competing interests.","order":1,"name":"Ethics","label":"Competing interests","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"This study was conducted in accordance with the Declaration of Helsinki and approved by the Ethics Committee of the Medical University of Vienna (EK418\/2009 - amendment). All patients provided informed consent before inclusion.","order":2,"name":"Ethics","label":"Ethics approval and consent to participate","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"ChatGPT 5.4 was used for language editing and formatting of the manuscript. The authors reviewed and approved all changes and take full responsibility for the final content of the manuscript.","order":3,"name":"Ethics","label":"AI-assisted manuscript preparation","group":{"name":"EthicsHeading","label":"Declarations"}}],"article-number":"33088"}}