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European journal of histochemistry : EJH2019; 63(4); doi: 10.4081/ejh.2019.3071

Bleaching melanin in formalin-fixed and paraffin-embedded melanoma specimens using visible light: a pilot study.

Abstract: In fluorescence microscopy, light radiation can be used to bleach fluorescent molecules in formalin-fixed and paraffin-embedded (FFPE) samples, in order to increase the ratio between signal of interest and background autofluorescence. We tested if the same principle can be exploited in bright field microscopy to bleach pigmented melanoma FFPE sections together with cell morphology maintenance. After dewaxing and rehydration, serial FFPE sections of a feline diffuse iris melanoma, a canine dermal melanoma, a gray horse dermal melanoma and a swine cutaneous melanoma were irradiated with visible light for 1, 2, 3, 4 and 5 days, prior to Hematoxylin and Eosin staining. Complete bleaching was obtained after 1-day treatment in feline and swine melanomas, while 2 and 3 days were required in canine and equine neoplasms, respectively. In all treated samples, cell morphology was maintained. Photo-induced bleaching combined with immunohistochemistry was tested after a 3-day photo-treatment using five different markers. According to the literature, in all samples neoplastic cells stained positive for vimentin, S100 and PNL2, while negative for FVIII and pancytokeratin. In conclusion, visible light can be effectively exploited to bleach pigmented melanoma FFPE sections prior to perform routine histochemical and immunohistochemical stains.
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Publication Date: 2019-10-31 PubMed ID: 31833329PubMed Central: PMC6829521DOI: 10.4081/ejh.2019.3071Google Scholar: Lookup
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  • Journal Article

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study explores the use of visible light to bleach melanin in formalin-fixed and paraffin-embedded (FFPE) melanoma samples for better microscopy results. The researchers established that sufficient light exposure, with durations varying between species, effectively bleached melanin, while preserving cell structure and compatibility with immunohistochemistry.

Research Methodology

  • The researchers used samples including a feline diffuse iris melanoma, a canine dermal melanoma, a gray horse dermal melanoma, and a swine cutaneous melanoma. These samples were formalin-fixed and paraffin-embedded (FFPE).
  • Following the process of dewaxing and rehydration, these FFPE sections were exposed to visible light for periods ranging from 1 to 5 days. This was done prior to staining them with Hematoxylin and Eosin

Observations and Findings

  • After 1-day light treatment, complete bleaching was achieved in the feline and swine melanomas.
  • For the canine and equine melanomas, bleaching required 2 to 3 days respectively.
  • In all cases, the cellular morphology was preserved which is crucial for maintaining the integrity of the samples for further analysis.
  • Besides, the microscopic analysis could be performed using bright field microscopy as well.

Immunohistochemistry Post Bleaching

  • Following a 3-day light exposure, the researchers performed immunohistochemistry using five different markers.
  • The neoplastic cells in all samples were found to test positive for vimentin, S100, and PNL2, and negative for FVIII and pancytokeratin, according to existing literature.

Conclusion

  • Overall, the study validates the use of visible light to effectively bleach pigmented melanoma FFPE sections.
  • This method not only preserves the cell morphology but also facilitates further histochemical and immunohistochemical stains, opening up opportunities for better visualization and diagnostic potential for melanomas.

Cite This Article

APA
Pigoli C, Gibelli LR, Caniatti M, Moretti L, Sironi G, Giudice C. (2019). Bleaching melanin in formalin-fixed and paraffin-embedded melanoma specimens using visible light: a pilot study. Eur J Histochem, 63(4). https://doi.org/10.4081/ejh.2019.3071

Publication

European journal of histochemistry : EJH
ISSN: 2038-8306
NlmUniqueID: 9207930
Country: Italy
Language: English
Volume: 63
Issue: 4

Researcher Affiliations

Pigoli, Claudio
  • Department of Veterinary Medicine (DIMEVET), University of Milan, Lodi. claudio.pigoli@unimi.it.
Gibelli, Lucia Rita
    Caniatti, Mario
      Moretti, Luca
        Sironi, Giuseppe
          Giudice, Chiara

            MeSH Terms

            • Animals
            • Antibodies, Monoclonal, Murine-Derived / immunology
            • Cats
            • Dogs
            • Formaldehyde / chemistry
            • Goats
            • Horses
            • Immunohistochemistry
            • Light
            • Melanins / radiation effects
            • Melanoma / pathology
            • Melanoma / veterinary
            • Mice
            • Paraffin Embedding
            • Photobleaching
            • Pilot Projects
            • Rabbits
            • S100 Proteins / immunology
            • Swine
            • Temperature
            • Vimentin / immunology

            Conflict of Interest Statement

            Conflict of interest: the authors declare they have no competing interests.

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            Citations

            This article has been cited 3 times.
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            3. Pellicciari C. Twenty years of histochemistry in the third millennium, browsing the scientific literature. Eur J Histochem 2020 Dec 29;64(4).
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