Semi–automated standardisation of melanin bleaching procedures of heavily pigmented melanocytic lesions for immunohistochemical analysis on an automated platform

Orchard, Guy; Gabriel, J; Shams, Mohammad Ghasem; Fernando, P; Satoc, J.; Nwokie, T; Ismail, Farzana; D’Amico, Cosimo

doi:10.1080/09674845.2019.1624083

Cited by 4 publications

(4 citation statements)

References 7 publications

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“…Diagnosis of malignant melanoma and other melanophilic tissues is frustrated by the possibility that over-pigmentation can mask crucial antibodyantigen reactions using di-amino benzidine. Orchard and colleague presented a method for reducing heavy pigmentation, so enabling more rapid and accurate diagnosis [32]. The same group also reported an advance in methods for screening for malignant melanoma, the genetic basis of many being a mutation in BRAF, which can be detected by a DNA probe.…”

Section: Cell Pathology and Molecular Geneticsmentioning

confidence: 99%

British Journal of Biomedical Science in 2019. What have we learned?

Blann

2019

British Journal of Biomedical Science

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In 2019 the British Journal of Biomedical Science published 40 articles in the various disciplines that comprise biomedical science. These were one review, 22 original articles and 17 'In Brief' short reports. Of those citing original data, the majority were in cellular pathology (14 papers), clinical chemistry (9 papers), and microbiology (6 papers: 4 in bacteriology and 2 in virology). There were 3 papers in haematology and 2 in andrology, whilst 5 papers crossed traditional discipline boundaries (such as the molecular genetics of IL6, liver function tests, and hepatocellular carcinoma). Over two-thirds of papers used techniques in molecular genetics. The present report will summarise key aspects of these publications that are of greatest relevance to laboratory scientists.

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Section: Cell Pathology and Molecular Geneticsmentioning

confidence: 99%

British Journal of Biomedical Science in 2019. What have we learned?

Blann

2019

British Journal of Biomedical Science

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“…Recently, some laboratories have developed semi-automatic or automated depigmentation and staining procedures through a series of automated standard operating modes, which have improved the quality of staining results and reduced manual errors. [16][17][18] In this study, diluted H 2 O 2 was placed in a fully automatic immunostaining machine, and then the experimental conditions were set to decolorize the melanin for histological staining and IHC. The purpose of this study was to observe whether melanin could be completely depigmented by oxidizing agents (H 2 O 2 ) without disrupting tissue morphology and antibody expression.…”

Section: Introductionmentioning

confidence: 99%

An Optimized Melanin Depigmentation Method for Histopathological and Immunohistochemical Staining of Formalin-Fixed Paraffin-Embedded Tissues

et al. 2023

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Introduction. The difficulty in diagnosis of severe melanocytic lesions is a problem to be overcome in pathological practice. Melanin bleaching is an effective approach to ameliorate melanin disturbances in severely pigmented lesions. Although various methods for improving melanin pigmentation in immunohistochemical staining have been reported, these depigmentation methods still need to be optimized and standardized. In this study, the coloring efficiency of 3,3′-diaminobenzidine (DAB) and alkaline phosphatase (AP) after melanin depigmentation was compared under the automatic immunohistochemical staining platform. Methods. The applicability of the optimized depigmentation method was validated in 10 formalin-fixed paraffin-embedded (FFPE) blocks of ocular melanoma tissues. Specimens were demelaninized with 10% hydrogen peroxide at 60°C for immunohistochemical staining (Melan-A and SOX10), and tissue chromogenic staining was performed with DAB and AP detection systems, respectively. Results. The optimized depigmentation method including immunohistochemistry (IHC) could be completed in 3 h, effectively preserving cell morphology and immunoreactivity. Among these, the color-rendering effect and contrast of AP are better than DAB. Conclusion. This optimized method can effectively remove melanin and improve the accuracy of IHC staining interpretation. AP staining has better visibility and readability without the interference of residual melanin. The comparison results showed that after melanin depigmentation, the immunohistochemical staining agent was replaced with red AP, which avoided the misjudgment caused by brown DAB when melanin depigmentation was incomplete. This improved method can be applied to future histopathological and immunohistochemical staining of melanin-deposited tissues.

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“…Strong oxidants, such as hydrogen peroxide and potassium permanganate (KMnO 4 ), are conventional depigmentation agents used in pathology laboratories 5–7 . Nevertheless, hydrogen peroxide bleaching is time-consuming (12 to 48 h) 8 .…”

mentioning

confidence: 99%

“…Strong oxidants, such as hydrogen peroxide and potassium permanganate (KMnO 4 ), are conventional depigmentation agents used in pathology laboratories. [5][6][7] Nevertheless, hydrogen peroxide bleaching is time-consuming (12 to 48 h). 8 Despite being a remarkable melanin remover, KMnO 4 damages the antigenicity of multiple tissue antigens, thus affecting the IHC staining and disease diagnosis.…”

mentioning

confidence: 99%

Depigmentation of Melanin-containing Tissues Using Hypochlorous Acid to Enhance Hematoxylin-eosin and Immunohistochemical Staining

Wang,

Wang

2023

Applied Immunohistochemistry &Amp; Molecular Morphology

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Pathologists diagnose diseases by observing the histologic and cellular morphology microscopically. However, the high pigmentation in melanin-containing tumors can hide the tumor cell structures, making diagnosing challenging. Previously, hydrogen peroxide and potassium permanganate were utilized for melanin bleaching with several limitations. For instance, hydrogen peroxide has a weak bleaching ability, and the process is time-consuming (12 h). Meanwhile, potassium permanganate affects the antigenicity of antigens and is unsuitable for immunohistochemical (IHC) staining. In this study, the hypochlorous acid (HClO) solution was applied to hematoxylin-eosin and IHC staining of melanin tissue sections. The study discovered that 1% HClO could completely bleach melanin particles in tumor tissues in a short period (19.95 ± 2.53 min) without compromising the hematoxylin-eosin staining. In addition, 2% HClO was utilized for bleaching at room temperature for 61.17 ± 4.32 minutes after the tissue was incubated with 3,3’-diaminobenzidine in IHC staining. This treatment effectively removed melanin without negatively impacting 3,3’-diaminobenzidine signal expression, thus ensuring that the sections met the necessary diagnostic requirements. Therefore, this method could facilitate pathologists in disease diagnosis of melanin-containing tissues.

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Semi–automated standardisation of melanin bleaching procedures of heavily pigmented melanocytic lesions for immunohistochemical analysis on an automated platform

Cited by 4 publications

References 7 publications

British Journal of Biomedical Science in 2019. What have we learned?

British Journal of Biomedical Science in 2019. What have we learned?

An Optimized Melanin Depigmentation Method for Histopathological and Immunohistochemical Staining of Formalin-Fixed Paraffin-Embedded Tissues

Depigmentation of Melanin-containing Tissues Using Hypochlorous Acid to Enhance Hematoxylin-eosin and Immunohistochemical Staining

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