{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,15]],"date-time":"2026-08-15T09:46:51Z","timestamp":1786787211193,"version":"build-2736575974"},"reference-count":12,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2011,5,4]],"date-time":"2011-05-04T00:00:00Z","timestamp":1304467200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pathology International"],"published-print":{"date-parts":[[2011,6]]},"abstract":"<jats:p>Excessive amounts of melanin pigments may hamper histopathological assessments of melanocytic lesions by obscuring cellular morphology and hindering antibody\u2013antigen interactions. To determine the optimal melanin\u2010bleaching conditions for histopathological examination, heavily pigmented melanomas were treated with warm hydrogen peroxide (H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>) diluted with various diluents (1% disodium hydrogen phosphate 12H<jats:sub>2<\/jats:sub>O ( (Na<jats:sub>2<\/jats:sub> HPO<jats:sub>4<\/jats:sub>); phosphate buffer 0.05\u2003M, pH 7.4 (PB); and PBS 0.05\u2003M, pH 7.4) at varying temperatures (50\u00b0C, 55\u00b0C, and 60\u00b0C) and for varying incubation times (0.5, 1, 2, and 3\u2003h). The effect of the sequential order of antigen retrieval and bleaching on preserving tissue morphology was then evaluated. Additionally, the effect of melanin bleaching using warm diluted H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> on the antigenicity of melanoma\u2010related markers (HMB\u201045, MART\u20101, and S\u2010100) and other markers used for histopathology was examined in amelanotic melanomas and tonsil tissue. Optimal and complete bleaching was achieved using warm 3% H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> in PB treatment at 55\u00b0C for 2\u2003h following antigen retrieval with microwaving or digestion with trypsin. Under these conditions, the tissue morphology and antigenicity of various immunohistochemical markers were also well preserved. Bleaching with warm 3% H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> PB is a fast and efficient method of bleaching melanin pigments and performing immunohistochemical examination in heavily melanin\u2010pigmented lesions.<\/jats:p>","DOI":"10.1111\/j.1440-1827.2011.02667.x","type":"journal-article","created":{"date-parts":[[2011,5,4]],"date-time":"2011-05-04T08:58:55Z","timestamp":1304499535000},"page":"345-350","source":"Crossref","is-referenced-by-count":33,"title":["Re\u2010evaluation of melanin bleaching using warm diluted hydrogen peroxide for histopathological analysis"],"prefix":"10.1111","volume":"61","author":[{"given":"Masanobu","family":"Momose","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiroyoshi","family":"Ota","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masayoshi","family":"Hayama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2011,5,4]]},"reference":[{"key":"e_1_2_6_2_2","first-page":"170","article-title":"The immunoalkaline phosphatase technique in immunohistochemistry: The effect of permanganate\u2010oxalate melanin bleaching upon four final reaction products","volume":"53","author":"Alexander RA","year":"1996","journal-title":"Br J Biomed Sci"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1600-0560.1991.tb01381.x"},{"key":"e_1_2_6_4_2","first-page":"1143","article-title":"A comparison of melanin bleaching and azure blue counterstaining in the immunohistochemical diagnosis of malignant melanoma","volume":"12","author":"Kligora CJ","year":"1999","journal-title":"Mod Pathol"},{"key":"e_1_2_6_5_2","first-page":"121","article-title":"Effect of melanin bleaching on immunoperoxidase, with reference to ocular tissues and lesions","volume":"43","author":"Alexander RA","year":"1986","journal-title":"Med Lab Sci"},{"key":"e_1_2_6_6_2","doi-asserted-by":"crossref","first-page":"480","DOI":"10.1093\/ajcp\/88.4.480","article-title":"The effect of melanin pigment removal on the peroxidase\u2010antiperoxidase immunoperoxidase technic","volume":"88","author":"McGovern J","year":"1987","journal-title":"Am J Clin Pathol"},{"key":"e_1_2_6_7_2","first-page":"22","article-title":"Immunohistochemical techniques: The effect of melanin bleaching","volume":"52","author":"Foss AJ","year":"1995","journal-title":"Br J Biomed Sci"},{"key":"e_1_2_6_8_2","first-page":"123","article-title":"Ueber die bleichung des melanins","volume":"19","author":"Strauss A","year":"1932","journal-title":"Zeitshrift fur Wissenschaftliche Microskopie fur mikroskopische Technik"},{"key":"e_1_2_6_9_2","first-page":"325","article-title":"Immunohistochemical staining followed by bleaching of melanin: A practical method for ophthalmic pathology","volume":"52","author":"Kivela T","year":"1995","journal-title":"Br J Biomed Sci"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1023\/A:1003700224900"},{"key":"e_1_2_6_11_2","first-page":"188","article-title":"Heavily pigmented melanocytic neoplasms: Comparison of two melanin\u2010bleaching techniques and subsequent immunohistochemical staining","volume":"56","author":"Orchard GE","year":"1999","journal-title":"Br J Biomed Sci"},{"key":"e_1_2_6_12_2","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1080\/09674845.2007.11978097","article-title":"Use of heat provides a fast and efficient way to undertake melanin bleaching with dilute hydrogen peroxide","volume":"64","author":"Orchard GE","year":"2007","journal-title":"Br J Biomed Sci"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1097\/00000372-199808000-00006"}],"container-title":["Pathology International"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fj.1440-1827.2011.02667.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/j.1440-1827.2011.02667.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,31]],"date-time":"2023-10-31T05:31:17Z","timestamp":1698730277000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/j.1440-1827.2011.02667.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,5,4]]},"references-count":12,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2011,6]]}},"alternative-id":["10.1111\/j.1440-1827.2011.02667.x"],"URL":"https:\/\/doi.org\/10.1111\/j.1440-1827.2011.02667.x","archive":["Portico"],"relation":{},"ISSN":["1320-5463","1440-1827"],"issn-type":[{"value":"1320-5463","type":"print"},{"value":"1440-1827","type":"electronic"}],"subject":[],"published":{"date-parts":[[2011,5,4]]}}}