{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T22:26:26Z","timestamp":1792794386580,"version":"4.5.0"},"reference-count":47,"publisher":"Oxford University Press (OUP)","issue":"7","license":[{"start":{"date-parts":[[2021,5,6]],"date-time":"2021-05-06T00:00:00Z","timestamp":1620259200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Oxford Medical Research Council Doctoral Training Programme","award":["MR\/N013468\/1"],"award-info":[{"award-number":["MR\/N013468\/1"]}]},{"name":"Fondation Hoffmann supporting R.A and the Petroleum Technology Development Fund","id":[{"id":"https:\/\/ror.org\/03zj57g67","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2021,6,18]]},"abstract":"Abstract<\/jats:title>\n \n STUDY QUESTION<\/jats:title>\n Can ovarian tissue morphology be better preserved whilst enabling histological molecular analyses following fixation with a novel fixative, neutral buffered formalin (NBF) with 5% acetic acid (referred to hereafter as Form-Acetic)?<\/jats:p>\n <\/jats:sec>\n \n SUMMARY ANSWER<\/jats:title>\n Fixation with Form-Acetic improved ovarian tissue histology compared to NBF in multiple species while still enabling histological molecular analyses.<\/jats:p>\n <\/jats:sec>\n \n WHAT IS KNOWN ALREADY<\/jats:title>\n NBF fixation results in tissue shrinkage in various tissue types including the ovary. Components of ovarian tissue, notably follicles, are particularly susceptible to NBF-induced morphological alterations and can lead to data misrepresentation. Bouin\u2019s solution (which contains 5% acetic acid) better preserves tissue architecture compared to NBF but is limited for immunohistochemical analyses.<\/jats:p>\n <\/jats:sec>\n \n STUDY DESIGN, SIZE, DURATION<\/jats:title>\n A comparison of routinely used fixatives, NBF and Bouin\u2019s, and a new fixative, Form-Acetic was carried out. Ovarian tissue was used from three different species: human (n\u2009=\u20095 patients), sheep (n\u2009=\u20093; 6 ovaries; 3 animals per condition) and mouse (n\u2009=\u200914 mice; 3 ovaries from 3 different animals per condition).<\/jats:p>\n <\/jats:sec>\n \n PARTICIPANTS\/MATERIALS, SETTING, METHODS<\/jats:title>\n Ovarian tissue from humans (aged 13\u2009weeks to 32\u2009years), sheep (reproductively young i.e. 3\u20136\u2009months) and mice (10\u2009weeks old) were obtained and fixed in 2\u2009ml NBF, Bouin\u2019s or Form-Acetic for 4, 8, and 24\u2009h at room temperature. Tissues were embedded and sectioned. Five-micron sections were stained with haemotoxylin and eosin (H&E) and the percentage of artefact (clear space as a result of shrinkage) between ovarian structures was calculated. Additional histological staining using Periodic acid-Schiff and Masson\u2019s trichrome were performed on 8 and 24\u2009h NBF, Bouin\u2019s and Form-Acetic fixed samples to assess the compatibility of the new fixative with stains. On ovarian tissue fixed for both 8 and 24\u2009h in NBF and Form-Acetic, immunohistochemistry (IHC) studies to detect FOXO3a, FoxL2, collagen IV, laminin and anti-M\u00fcllerian hormone (AMH) proteins were performed in addition to the terminal deoxynucleotidyl transferase nick end labelling (TUNEL) assay to determine the compatibility of Form-Acetic fixation with types of histological molecular analyses.<\/jats:p>\n <\/jats:sec>\n \n MAIN RESULTS AND THE ROLE OF CHANCE<\/jats:title>\n Fixation in Form-Acetic improved ovarian tissue morphology compared to NBF from all three species and either slightly improved or was comparable to Bouin\u2019s for human, mouse and sheep tissues. Form-Acetic was compatible with H&E, Periodic acid-Schiff and Masson\u2019s trichrome staining and all proteins (FOXO3a, FoxL2, collagen IV and laminin and AMH) could be detected via IHC. Furthermore, Form-Acetic, unlike NBF, enabled antigen recognition for most of the proteins tested without the need for antigen retrieval. Form-Acetic also enabled the detection of damaged DNA via the TUNEL assay using fluorescence.<\/jats:p>\n <\/jats:sec>\n \n LARGE SCALE DATA<\/jats:title>\n N\/A<\/jats:p>\n <\/jats:sec>\n \n LIMITATIONS, REASONS FOR CAUTION<\/jats:title>\n In this study, IHC analysis was performed on a select number of protein types in ovarian tissue thus encouraging further studies to confirm the use of Form-Acetic in enabling the detection of a wider range of protein forms in addition to other tissue types.<\/jats:p>\n <\/jats:sec>\n \n WIDER IMPLICATIONS OF THE FINDINGS<\/jats:title>\n The simplicity in preparation of Form-Acetic and its superior preservative properties whilst enabling forms of histological molecular analyses make it a highly valuable tool for studying ovarian tissue. We, therefore, recommend that Form-Acetic replaces currently used fixatives and encourage others to introduce it into their research workflow.<\/jats:p>\n <\/jats:sec>\n \n STUDY FUNDING\/COMPETING INTEREST(S)<\/jats:title>\n This work was supported by the Oxford Medical Research Council Doctoral Training Programme (Oxford MRC-DTP) grant awarded to B.D.B. (Grant no. MR\/N013468\/1), the Fondation Hoffmann supporting R.A. and the Petroleum Technology Development Fund (PTDF) awarded to B.V.A.<\/jats:p>\n <\/jats:sec>","DOI":"10.1093\/humrep\/deab075","type":"journal-article","created":{"date-parts":[[2021,3,26]],"date-time":"2021-03-26T00:13:29Z","timestamp":1616717609000},"page":"1871-1890","source":"Crossref","is-referenced-by-count":30,"title":["Improved preservation of ovarian tissue morphology that is compatible with antigen detection using a fixative mixture of formalin and acetic acid"],"prefix":"10.1093","volume":"36","author":[{"given":"B V","family":"Adeniran","sequence":"first","affiliation":[{"name":"Nuffield Department of Women\u2019s and Reproductive Health, Women\u2019s Centre, John Radcliffe Hospital, University of Oxford, Oxford, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B D","family":"Bjarkadottir","sequence":"additional","affiliation":[{"name":"Nuffield Department of Women\u2019s and Reproductive Health, Women\u2019s Centre, John Radcliffe Hospital, University of Oxford, Oxford, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0170-1963","authenticated-orcid":false,"given":"R","family":"Appeltant","sequence":"additional","affiliation":[{"name":"Nuffield Department of Women\u2019s and Reproductive Health, Women\u2019s Centre, John Radcliffe Hospital, University of Oxford, Oxford, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"S","family":"Lane","sequence":"additional","affiliation":[{"name":"Future Fertility Programme Oxford, Oxford, UK"},{"name":"Department of Paediatric Oncology and Haematology, Children's Hospital Oxford, Oxford University Hospitals NHS Foundation Trust, Oxford, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1798-976X","authenticated-orcid":false,"given":"S A","family":"Williams","sequence":"additional","affiliation":[{"name":"Nuffield Department of Women\u2019s and Reproductive Health, Women\u2019s Centre, John Radcliffe Hospital, University of Oxford, Oxford, UK"},{"name":"Future Fertility Programme Oxford, Oxford, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2021,5,6]]},"reference":[{"key":"2021070807484037800_deab075-B1","doi-asserted-by":"crossref","first-page":"312","DOI":"10.1016\/B978-0-407-00154-1.50017-8","volume-title":"Introduction to Medical Laboratory Technology","author":"Baker","year":"1976"},{"key":"2021070807484037800_deab075-B2","doi-asserted-by":"crossref","DOI":"10.5962\/bhl.title.5905","volume-title":"Principles of Biological Microtechnique; 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