ArticleLaboratory investigation; a journal of technical methods and pathology2016
A melanin-bleaching methodology for molecular and histopathological analysis of formalin-fixed paraffin-embedded tissue.
Article in Laboratory investigation; a journal of technical methods and pathology, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 29 citations in OpenAlex.
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- Mapping of individual sensory nerve axons from digits to spinal cord with the transparent embedding solvent system.Cell research · 2024Article
- TIPARP as a prognostic biomarker and potential immunotherapeutic target in male papillary thyroid carcinoma.Cancer cell international · 2024Article
- BRAFCancer cell international · 2023Article
- The Application of Guanidinium to Improve Biomolecule Quality in Fixed, Paraffin-embedded Tissue.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2023Article
- Protein and lipid mass concentration measurement in tissues by stimulated Raman scattering microscopy.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Histomorphological and Molecular Assessments of the Fixation Times Comparing Formalin and Ethanol-Based Fixatives.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2018Article
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Authors and funding
10 authors at 5 institutions in 2 countries.
Funding
Abstract
Removal 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.
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