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However, the standard frozen section analysis (FSA) procedure does not allow precise interpretation of the final histopathological results. For this reason, we have developed a modified intraoperative margins (IM) assessment protocol using virtual 3D images of resected OSCC.<\/jats:p><\/jats:sec><jats:sec><jats:title>Material and methods<\/jats:title><jats:p>The pilot study included 10 patients with OSCC located in the floor of the mouth (FOM, <jats:italic>n<\/jats:italic> = 3), tongue (TC, <jats:italic>n<\/jats:italic> = 2), maxilla (MT, <jats:italic>n<\/jats:italic> = 1), and retromolar triangle (RMT, <jats:italic>n<\/jats:italic> = 4). After excision, OSCCs were scanned and mapped, marked on the virtual preparation of the IM collection sites. The modified pathological assessment of the IM enabled the evaluation of dimensions, including the width of the collected IM.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>By analyzing virtual OSCC images, we demonstrated the effectiveness of a modified IM assessment methodology. In one case study, the oncological council disqualified the patient from adjuvant radiotherapy.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Leveraging the OSCC\u2019s virtual mapping capabilities and the modified IM pathology methodology, we developed an IM evaluation protocol that enables the incorporation of IM width into the OSCC\u2019s microscopic excision margin, thereby significantly enhancing the diagnostic value and precision of the final OSCC histopathology results.<\/jats:p><\/jats:sec>","DOI":"10.3389\/fmed.2025.1638833","type":"journal-article","created":{"date-parts":[[2025,9,29]],"date-time":"2025-09-29T05:27:08Z","timestamp":1759123628000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":1,"title":["Digital localization of intraoperative samples enhances margin analysis precision in oral squamous cell 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