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A melanin-bleaching methodology for molecular and histopathological analysis of formalin-fixed paraffin-embedded

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dc.contributor.author조한별-
dc.date.accessioned2017-10-26T07:35:59Z-
dc.date.available2017-10-26T07:35:59Z-
dc.date.issued2016-
dc.identifier.issn0023-6837-
dc.identifier.urihttps://ir.ymlib.yonsei.ac.kr/handle/22282913/152253-
dc.description.abstractRemoval of excessive melanin from heavily pigmented formalin-fixed paraffin-embedded (FFPE) melanoma tissues is essential for histomorphological and molecular diagnostic assessments. Although there have been efforts to address this issue, current methodologies remain complex and time-consuming, and are not suitable for multiple molecular applications. Herein, we have developed a robust and rapid melanin-bleaching methodology for FFPE tissue specimens. Our approach is based on quick bleaching (15?min) at high temperature (80?°C) with 0.5% diluted hydrogen peroxide (H2O2) in Tris-HCl, PBS, or Tris/Tricine/SDS buffer. Immunostaining for Ki-67 and HMB45 was enhanced by bleaching with 0.5% H2O2 in Tris/Tricine/SDS and Tris-HCl, respectively. In addition to histopathological applications, our approach also facilitates recovery of protein and nucleic acid from archival melanin-rich FFPE tissue sections. Protein extracted from bleached FFPE tissues was compatible with western blotting using anti-human GAPDH and AKT antibodies. Our bleaching condition significantly improved RNA quality compared with unbleached tissues without compromising the yield. Notably, the RNA/DNA obtained from bleached tissues was suitable for end point PCR and real-time quantitative RT-PCR. In conclusion, this improved melanin-bleaching method enhances and simplifies immunostaining procedures, and facilitates the use of melanin-rich FFPE tissues for histomorphological and PCR amplification-based molecular assays.-
dc.description.statementOfResponsibilityrestriction-
dc.languageEnglish-
dc.publisherNature Pub. Group-
dc.relation.isPartOfLABORATORY INVESTIGATION-
dc.rightsCC BY-NC-ND 2.0 KR-
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/2.0/kr/-
dc.titleA melanin-bleaching methodology for molecular and histopathological analysis of formalin-fixed paraffin-embedded-
dc.typeArticle-
dc.publisher.locationUnited States-
dc.contributor.collegeCollege of Medicine-
dc.contributor.departmentDept. of Obstetrics & Gynecology-
dc.contributor.googleauthorJoon-Yong Chung-
dc.contributor.googleauthorJiyeon Choi-
dc.contributor.googleauthorJohn D Sears-
dc.contributor.googleauthorKris Ylaya-
dc.contributor.googleauthorCandice Perry-
dc.contributor.googleauthorChel H Choi-
dc.contributor.googleauthorSeung-Mo Hong-
dc.contributor.googleauthorHanbyoul Cho-
dc.contributor.googleauthorKevin M Brown-
dc.contributor.googleauthorStephen M Hewitt-
dc.identifier.doi10.1038/labinvest.2016.90-
dc.contributor.localIdA03921-
dc.relation.journalcodeJ02150-
dc.identifier.eissn1530-0307-
dc.identifier.pmid27548802-
dc.identifier.urlhttp://www.nature.com/labinvest/journal/v96/n10/full/labinvest201690a.html-
dc.contributor.alternativeNameCho, Han Byoul-
dc.contributor.affiliatedAuthorCho, Han Byoul-
dc.citation.volume96-
dc.citation.number10-
dc.citation.startPage1116-
dc.citation.endPage1127-
dc.identifier.bibliographicCitationLABORATORY INVESTIGATION, Vol.96(10) : 1116-1127, 2016-
dc.date.modified2017-10-24-
dc.identifier.rimsid47990-
dc.type.rimsART-
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Obstetrics and Gynecology (산부인과학교실) > 1. Journal Papers

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