{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T20:34:22Z","timestamp":1792787662505,"version":"4.5.0"},"reference-count":27,"publisher":"SAGE Publications","issue":"4","license":[{"start":{"date-parts":[[2024,4,22]],"date-time":"2024-04-22T00:00:00Z","timestamp":1713744000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"name":"Zoological Pathology Program"},{"DOI":"10.13039\/100000026","name":"NIH","doi-asserted-by":"publisher","award":["T35 OD011145"],"award-info":[{"award-number":["T35 OD011145"]}],"id":[{"id":"10.13039\/100000026","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["J VET Diagn Invest"],"published-print":{"date-parts":[[2024,7]]},"abstract":"<jats:p>RNAscope in situ hybridization (ISH) detects target RNA in formalin-fixed, paraffin-embedded (FFPE) tissues. Protocols suggest that prolonged FFPE storage and formalin fixation may impact signal detection, potentially limiting the utility of RNAscope ISH in retrospective studies. To develop parameters for RNAscope use with archived specimens, we evaluated the effect of formalin-fixation time by measuring the signal of a reference gene ( 16srRNA) in selected tissues fixed in 10% neutral-buffered formalin for 1, 2, 3, 5, 7, 10, 14, 21, 28, 60, 90, 180, and 270\u2009d. The signal intensity and percent area of signal decreased after 180\u2009d. Tissues had detectable signal at 180\u2009d but not at 270\u2009d of formalin fixation. To assess target detection in paraffin blocks, we qualitatively compared the signal of canine distemper virus (CDV) antigen via immunohistochemistry and CDV RNA via RNAscope ISH in replicate sections from blocks stored at room temperature for 6 mo, 1, 3, 6, 8, 11, 13, and 15\u2009y; RNA was detected in FFPE tissues stored for up to 15\u2009y. Our results demonstrate that RNAscope ISH can detect targets in tissues with prolonged paraffin storage intervals and formalin-fixation times.<\/jats:p>","DOI":"10.1177\/10406387241245777","type":"journal-article","created":{"date-parts":[[2024,4,23]],"date-time":"2024-04-23T00:39:37Z","timestamp":1713832777000},"page":"498-505","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":7,"title":["Effect of formalin-fixation and paraffin-embedded tissue storage times on RNAscope in situ hybridization signal amplification"],"prefix":"10.1177","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6674-772X","authenticated-orcid":false,"given":"Megan E.","family":"Colburn","sequence":"first","affiliation":[{"name":"Zoological Pathology Program, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, IL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martha A.","family":"Delaney","sequence":"additional","affiliation":[{"name":"Zoological Pathology Program, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, IL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1910-1410","authenticated-orcid":false,"given":"Gretchen C.","family":"Anchor","sequence":"additional","affiliation":[{"name":"Zoological Pathology Program, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, IL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karen A.","family":"Terio","sequence":"additional","affiliation":[{"name":"Zoological Pathology Program, College of Veterinary Medicine, University of Illinois at Urbana-Champaign, IL, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2024,4,22]]},"reference":[{"key":"bibr1-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1177\/0300985818758470"},{"key":"bibr2-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/aqx094"},{"key":"bibr3-10406387241245777","doi-asserted-by":"publisher","DOI":"10.3201\/eid2107.150228"},{"key":"bibr4-10406387241245777","doi-asserted-by":"publisher","DOI":"10.18632\/oncotarget.21851"},{"key":"bibr5-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1177\/104063870902100510"},{"key":"bibr6-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1638\/2022-0042"},{"key":"bibr7-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1111\/vco.12825"},{"key":"bibr8-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1093\/toxsci\/kfw161"},{"key":"bibr9-10406387241245777","doi-asserted-by":"publisher","DOI":"10.1097\/PDM.0b013e3181e71ba5"},{"key":"bibr10-10406387241245777","unstructured":"International Union for Conservation of Nature and Natural Resources (IUCN) SSC Antelope Specialist Group. 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