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Current clinical diagnostics use electrical and structural markers, but lack precision due to low spatial resolution and absence of molecular information. In this paper, we employed line scan Raman microspectroscopy to classify sheep myocardial tissue into muscle, necrotic, granulated, and fibrotic tissue types, using collagen as a molecular biomarker. Three spectral regions were evaluated: region A (600\u20132960 cm\u22121), region B (600\u20131399 cm\u22121 and 1751\u20132960 cm\u22121), and region C (1400\u20131750 cm\u22121), which includes the prominent collagen-associated peaks at 1448 cm\u22121 and 1652 cm\u22121. Linear and nonlinear principal component analysis (PCA) and support vector machines (SVMs) were applied for dimensionality reduction and classification, with nonlinear models specifically addressing the nonlinearity of collagen formation during fibrogenesis. Histological validation was performed using Masson\u2019s trichrome staining. Raman bands associated with collagen in region C consistently outperformed regions A and B, achieving the highest explained variance and best class separation in both binary and multiclass PCA models for both linear and nonlinear approaches. The ratio of collagen-related peaks enabled stage-dependent tissue characterization, confirming the nonlinear nature of fibrotic remodeling. Our findings highlight the diagnostic potential of collagen-associated Raman bands for characterizing myocardial fibrosis. The proposed PCA-SVM framework demonstrates robust performance even with limited sample size and has the potential to lay the foundation for real-time intraoperative diagnostics.<\/jats:p>","DOI":"10.3390\/ijms26157240","type":"journal-article","created":{"date-parts":[[2025,7,28]],"date-time":"2025-07-28T10:51:14Z","timestamp":1753699874000},"page":"7240","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Spectral Region Optimization and Machine Learning-Based Nonlinear Spectral Analysis for Raman Detection of Cardiac Fibrosis Following Myocardial Infarction"],"prefix":"10.3390","volume":"26","author":[{"given":"Arno","family":"Krause","sequence":"first","affiliation":[{"name":"Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8212-0545","authenticated-orcid":false,"given":"Marco","family":"Andreana","sequence":"additional","affiliation":[{"name":"Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard D.","family":"Walton","sequence":"additional","affiliation":[{"name":"IHU Liryc, University of Bordeaux, INSERM U 1045, CRCTB, F-33000 Bordeaux, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9551-835X","authenticated-orcid":false,"given":"James","family":"Marchant","sequence":"additional","affiliation":[{"name":"IHU Liryc, University of Bordeaux, INSERM U 1045, CRCTB, F-33000 Bordeaux, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nestor","family":"Pallares-Lupon","sequence":"additional","affiliation":[{"name":"IHU Liryc, University of Bordeaux, INSERM U 1045, CRCTB, F-33000 Bordeaux, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3185-0484","authenticated-orcid":false,"given":"Kanchan","family":"Kulkarni","sequence":"additional","affiliation":[{"name":"IHU Liryc, University of Bordeaux, INSERM U 1045, CRCTB, F-33000 Bordeaux, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wolfgang","family":"Drexler","sequence":"additional","affiliation":[{"name":"Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1251-3001","authenticated-orcid":false,"given":"Angelika","family":"Unterhuber","sequence":"additional","affiliation":[{"name":"Center for Medical Physics and Biomedical Engineering, Medical University of Vienna, Waehringer Guertel 18-20, 1090 Vienna, Austria"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,7,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"6","DOI":"10.1016\/j.hlc.2018.08.026","article-title":"Epidemiology of Sudden Cardiac Death: Global and Regional Perspectives","volume":"28","author":"Wong","year":"2019","journal-title":"Hear. 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