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Previous biomechanical analyses successfully discriminated post-mortem brain tissue from tissue with a post-mortem interval of at least one day when held at 20\u00a0\u00b0C. However, the practical utility of such analyses beyond day one at 20\u00a0\u00b0C was limited. This study investigates the storage, loss, and complex shear modulus of various brain regions in sheep stored at 4\u00a0\u00b0C in 24-hour intervals over four days post-mortem using rheometry tests. The aim is to identify the critical biomechanical tissue property values to predict post-mortem time and assess the temperature sensitivity of the rheometry method by comparing results to recent findings at 20\u00a0\u00b0C.\u00a0Thirty sheep brains were examined, including the frontal lobe, parietal lobe, anterior and posterior deep brain, superior colliculi, pons, medulla, and cerebellum. Rheometry tests were conducted, and receiver operator characteristic analyses were employed to establish cut-off values.\u00a0At 4\u00a0\u00b0C storage, all investigated biomechanical properties of the examined brain regions remained stable for at least one day post-mortem. Using cerebellar samples stored at 4\u00a0\u00b0C, a post-mortem interval of at least two days could be determined with excellent diagnostic ability. Complex shear modulus values below 1435\u00a0Pa or storage modulus values below 1313\u00a0Pa allowed prediction of two or more days post-mortem. Comparisons between 4\u00a0\u00b0C and 20\u00a0\u00b0C revealed brain region-specific results. For instance, the complex shear moduli of the anterior deep brain at 4\u00a0\u00b0C were significantly higher on all individual testing days when compared to 20\u00a0\u00b0C. In contrast, the combined medulla and pons samples were similar on each day.\u00a0Rheometry testing of brain tissue consistently stored at 4\u00a0\u00b0C since death proved valuable for forensic time since death estimations starting from two days after death.<\/jats:p>","DOI":"10.1007\/s00414-024-03267-3","type":"journal-article","created":{"date-parts":[[2024,6,20]],"date-time":"2024-06-20T19:01:40Z","timestamp":1718910100000},"page":"2541-2549","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["The influence of cooling on biomechanical time since death estimations using ovine brain tissue"],"prefix":"10.1007","volume":"138","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-3560-3915","authenticated-orcid":false,"given":"Johann","family":"Zwirner","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pavithran","family":"Devananthan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul D.","family":"Docherty","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Ondruschka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Natalia","family":"Kabaliuk","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,6,20]]},"reference":[{"key":"3267_CR1","doi-asserted-by":"publisher","first-page":"328","DOI":"10.1007\/s00194-022-00580-2","volume":"32","author":"J Zwirner","year":"2022","unstructured":"Zwirner J, Devananthan P, Docherty P, Ondruschka B, Kabaliuk N (2022) 'Forensic tissue mechanics' for time since death estimations \u2013 a proof of concept. 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