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To determine the best retrieval method for common microRNAs, a multiblock composed of paraffin-embedded tonsil, cervix, placenta, and hyperplastic prostate tissue were included. Tissue were fixed in 10% formalin in a range of 5\u2013144\u00a0hours (h). Cut sections (5\u00a0\u03bcm) from the\u00a0multiblock were subjected to combinations of pretreatment procedures: variable periods of proteinase K (PK) digestion or Heat-induced microRNA Retrieval (HmiRR) using target retrieval solution (TRS) pH 6.1 or 9, with or without enzymatic treatment (pepsin). Results for the\u00a0overall categories: TRS pH 9 versus PK; <jats:italic>p<\/jats:italic>\u2009=\u20092.9e\u221223, TRS pH 9 versus TRS pH 6.1; <jats:italic>p<\/jats:italic>\u2009=\u20091.1e\u221214, TRS pH 6.1 versus PK; <jats:italic>p<\/jats:italic>\u2009=\u20092.9e\u221203. A long fixation time, resulted in the best microRNA preservation and staining intensity (long vs. short: <jats:italic>p<\/jats:italic>\u2009=\u20093.5e\u221247, long vs. moderate: <jats:italic>p<\/jats:italic>\u2009=\u20091.6e\u221244, moderate vs. short: <jats:italic>p<\/jats:italic>\u2009=\u20094.3e\u221216), was enhanced using HmiRR TRS pH\u00a09 with or without pepsin providing high sensitivity and specificity. These observations conflict with other ISH techniques (e.g., messenger ribonucleic acid), which typically require shorter fixation periods, and therefore, further studies are warranted.<\/jats:p>","DOI":"10.1038\/s41598-021-83888-5","type":"journal-article","created":{"date-parts":[[2021,2,24]],"date-time":"2021-02-24T11:03:35Z","timestamp":1614164615000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["A novel approach for microRNA in situ hybridization using locked nucleic acid probes"],"prefix":"10.1038","volume":"11","author":[{"given":"Isabella W.","family":"Paulsen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Bzorek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesper","family":"Olsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Birgitte","family":"Grum-Schwensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jesper T.","family":"Troelsen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ole B.","family":"Pedersen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2021,2,24]]},"reference":[{"key":"83888_CR1","doi-asserted-by":"publisher","first-page":"1813","DOI":"10.1101\/gad.5.10.1813","volume":"5","author":"B Wightman","year":"1991","unstructured":"Wightman, B., B\u00fcrglin, T. 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