{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,18]],"date-time":"2026-08-18T02:55:21Z","timestamp":1787021721800,"version":"build-2736575974"},"reference-count":33,"publisher":"SAGE Publications","issue":"6","license":[{"start":{"date-parts":[[2021,5,19]],"date-time":"2021-05-19T00:00:00Z","timestamp":1621382400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"funder":[{"DOI":"10.13039\/100011965","name":"integrated biobank of luxembourg","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100011965","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["J Histochem Cytochem."],"published-print":{"date-parts":[[2021,6]]},"abstract":"<jats:p>\n                    We evaluate the consequences of processing alcohol-fixed tissue in a processor previously used for formalin-fixed tissue. Biospecimens fixed in PAXgene Tissue Fixative were cut into three pieces then processed in a flushed tissue processor previously used for formalin-fixed, paraffin-embedded (FFPE) blocks (neutral buffered formalin [NBF]\n                    <jats:sup>+ve<\/jats:sup>\n                    ), a formalin-free system (NBF\n                    <jats:sup>\u2212ve<\/jats:sup>\n                    ), or left unprocessed. Histomorphology and immunohistochemistry were compared using hematoxylin\/eosin staining and antibodies for MLH-1, Ki-67, and CK-7. Nucleic acid was extracted using the PAXgene Tissue RNA\/DNA kits and an FFPE RNA extraction kit. RNA integrity was assessed using RNA integrity number (RIN), reverse transcription polymerase chain reaction (RT-PCR) (four amplicons), and quantitative RT-PCR (three genes). For DNA, multiplex PCR, quantitative PCR, DNA integrity number, and gel electrophoresis were used. Compared with NBF\n                    <jats:sup>\u2212ve<\/jats:sup>\n                    , RNA from NBF\n                    <jats:sup>+ve<\/jats:sup>\n                    blocks had 88% lower yield and poorer purity; average RIN reduced from 5.0 to 3.8, amplicon length was 408 base pairs shorter, and Cq numbers were 1.9\u20132.4 higher. Using the FFPE extraction kit rescued yield and purity, but RIN further declined by 1.1 units. Differences between NBF\n                    <jats:sup>+ve<\/jats:sup>\n                    and NBF\n                    <jats:sup>\u2212ve<\/jats:sup>\n                    in respect of DNA, histomorphology, and immunohistochemistry were either non-existent or small in magnitude. Formalin contamination of a tissue processor and its reagents therefore critically reduce RNA yield and integrity. We discuss the available options users can adopt to ameliorate this problem:\n                  <\/jats:p>","DOI":"10.1369\/00221554211017859","type":"journal-article","created":{"date-parts":[[2021,5,19]],"date-time":"2021-05-19T13:21:57Z","timestamp":1621430517000},"page":"389-405","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":5,"title":["Do Tissues Fixed in a Non-crosslinking Fixative Require a Dedicated Formalin-free Processor?"],"prefix":"10.1369","volume":"69","author":[{"given":"Sonia G.","family":"Frasquilho","sequence":"first","affiliation":[{"name":"Integrated Biobank of Luxembourg, Dudelange, Luxembourg"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ignacio","family":"Sanchez","sequence":"additional","affiliation":[{"name":"Integrated Biobank of Luxembourg, Dudelange, Luxembourg"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Changyoung","family":"Yoo","sequence":"additional","affiliation":[{"name":"The Catholic University of Korea, St. Vincent\u2019s Hospital, Seoul, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laurent","family":"Antunes","sequence":"additional","affiliation":[{"name":"Laboratoire National de Sant\u00e9, Dudelange, Luxembourg"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Camille","family":"Bellora","sequence":"additional","affiliation":[{"name":"Integrated Biobank of Luxembourg, Dudelange, Luxembourg"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"William","family":"Mathieson","sequence":"additional","affiliation":[{"name":"Integrated Biobank of Luxembourg, Dudelange, Luxembourg"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"179","published-online":{"date-parts":[[2021,5,19]]},"reference":[{"key":"bibr1-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1177\/33.8.3894502"},{"issue":"7156","key":"bibr2-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1038\/448959a","volume":"448","author":"Blow N.","year":"2007","journal-title":"Nature"},{"key":"bibr3-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1038\/gim.2017.241"},{"key":"bibr4-00221554211017859","doi-asserted-by":"publisher","DOI":"10.3390\/ijms18020308"},{"issue":"5","key":"bibr5-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1080\/10520295.2018.1446101","volume":"93","author":"Gaffney EF","year":"2018","journal-title":"Biotech Histochem"},{"key":"bibr6-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-5347(17)53865-3"},{"key":"bibr7-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1097\/00007611-198509000-00007"},{"issue":"6","key":"bibr8-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1016\/j.anndiagpath.2008.07.004","volume":"12","author":"Buesa RJ","year":"2008","journal-title":"Annals of Diagnostic Pathology"},{"issue":"5","key":"bibr9-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1016\/j.jmoldx.2012.05.002","volume":"14","author":"Viertler C","year":"2012","journal-title":"J Mol Diagn"},{"key":"bibr10-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/aqy026"},{"key":"bibr11-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1089\/bio.2015.29031.hmm"},{"key":"bibr12-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1093\/ajcp\/aqw023"},{"key":"bibr13-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1089\/bio.2015.0008"},{"key":"bibr14-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1038\/sj.bjc.6602889"},{"issue":"4","key":"bibr15-00221554211017859","volume":"9","author":"Wang FF","year":"2007","journal-title":"J Mol Diagn"},{"key":"bibr16-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1089\/bio.2018.0009","volume":"16","author":"Mathieson W","year":"2018","journal-title":"Biopreserv Biobank"},{"key":"bibr17-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1016\/j.cancergen.2015.05.001"},{"key":"bibr18-00221554211017859","unstructured":"PreAnalytiX. Technical note: influence of formalin contamination during processing of PAXgene tissue fixed, paraffin-embedded tissue (PFPE) on RNA yield, integrity, and performance in quantitative RT-PCR 2010. Available from: https:\/\/www.preanalytix.com\/storage\/download\/_ProductResources_\/TechnicalNotes\/PROM-2102-002_TN_Tissue_RNA_System_Influence_of_formalin_contamination_during_PFPE_tissue_processing_on_RNA_yield_integrity_amplification_0610_WW.pdf"},{"issue":"1","key":"bibr19-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1097\/PAS.0000000000000961","volume":"42","author":"Hognas G","year":"2018","journal-title":"Am J Surg Pathol"},{"issue":"8","key":"bibr20-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1016\/j.prp.2015.11.008","volume":"212","author":"Belder N","year":"2016","journal-title":"Pathol Res Pract"},{"issue":"3","key":"bibr21-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1097\/01.pdm.0000213450.99655.54","volume":"15","author":"Koch I","year":"2006","journal-title":"Diagn Mol Pathol"},{"issue":"1","key":"bibr22-00221554211017859","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1111\/j.1556-4029.2009.01245.x","volume":"55","author":"Opel KL","year":"2010","journal-title":"J Forensic Sci"},{"key":"bibr23-00221554211017859","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.jcmgh.2020.07.010","volume":"11","author":"Dotsenko V","year":"2020","journal-title":"Cell Mol Gastroenterol Hepatol"},{"issue":"9","key":"bibr24-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1136\/jclinpath-2012-201283","volume":"66","author":"Staff S","year":"2013","journal-title":"J Clin Pathol"},{"key":"bibr25-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1186\/s12876-019-1089-7"},{"key":"bibr26-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1002\/cjp2.145"},{"issue":"1","key":"bibr27-00221554211017859","volume":"42","author":"Liu Y","year":"2017","journal-title":"Curr Eye Res"},{"issue":"10","key":"bibr28-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1021\/pr100664e","volume":"9","author":"Ergin B","year":"2010","journal-title":"J Proteome Res"},{"issue":"3","key":"bibr29-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1293\/tox.2017-0049","volume":"31","author":"Yamazaki M","year":"2018","journal-title":"J Toxicol Pathol"},{"issue":"5","key":"bibr30-00221554211017859","doi-asserted-by":"crossref","DOI":"10.1089\/bio.2014.0101","volume":"13","author":"Hara K","year":"2015","journal-title":"Biopreserv Biobank"},{"key":"bibr31-00221554211017859","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0236104"},{"key":"bibr32-00221554211017859","unstructured":"Agilent Technologies. 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