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Many protocols for TEM sample preparation of collagen involve one or more washing steps to remove excess salts from the dispersion that could hamper analysis when dried on a TEM grid. Such protocols are not standardized and washing times as well as washing solvents vary from procedure to procedure, with each research group typically having their own protocol. Here, we investigate the influence of washing with water, ethanol, but also methanol and 2\u2010propanol, for both mineralized and unmineralized collagen samples via a protocol based on centrifugation. Washing with water maintains the hydrated collagen structure and the characteristic banding pattern can be clearly observed. Conversely, washing with ethanol results in dehydration of the fibrils, often leading to aggregation of the fibers and a less obvious banding pattern, already within 1\u00a0min of ethanol exposure. As we show, this process is fully reversible. Similar observations were made for methanol and propanol. Based on these results, a standardized washing protocol for collagenous samples is proposed.<\/jats:p>","DOI":"10.1002\/jemt.23915","type":"journal-article","created":{"date-parts":[[2021,8,27]],"date-time":"2021-08-27T07:35:53Z","timestamp":1630049753000},"page":"412-417","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["The effects of washing a collagen sample prior to <scp>TEM<\/scp> examination"],"prefix":"10.1002","volume":"85","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8083-2581","authenticated-orcid":false,"given":"Bernette Maria","family":"Oosterlaken","sequence":"first","affiliation":[{"name":"Laboratory of Physical Chemistry, Department of Chemical Engineering and Chemistry Eindhoven University of Technology  Eindhoven Netherlands"},{"name":"Center for Multiscale Electron Microscopy Eindhoven University of Technology  Eindhoven 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