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Influence of decalcifying agents on immunoreactivity of formalin-fixed, paraffin-embedded tissue

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Summary

The influence of eight decalcifying agents on the immunoreactivity of formalin-fixed, paraffin-embedded tissue for immunoglobulins, lysozyme, factor VIII-related antigen and keratin was studied using the unlabelled antibody peroxidase-antiperoxidase (PAP) method. Limited studies were also performed on tissues fixed in acid-formalin mixtures. All tissues were stained using an indirect immunoperoxidase method with mouse monoclonal antibodies to IgM, kappa and lambda light chains. The results suggest that, with controlled trypsinization of sections, it was possible to obtain optimal immunostaining for all tested antigens, with adequate preservation of histological structure, after decalcification in neutral EDTA or 10% aqueous acetic or formic acid. Tissue treated with agents containing mineral acids exhibited variable immunoreactivity and impaired counterstaining.

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References

  • Banks, P. M. (1979) Diagnostic applications of an immunoperoxidase method in hematopathology.J. Histochem. Cytochem. 24, 1192–4.

    Google Scholar 

  • Bosman, F. T., Lindeman, J., Kuiper G., Van Der Wal, A. &Kreunig, J. (1977) The influence of fixation on immunoperoxidase staining of plasma cells in paraffin sections of intestinal biopsy specimens.Histochemistry 53, 57–62.

    Google Scholar 

  • Brandtzaeg, P. (1983) Tissue preparation methods for immunohistochemistry. In:Techniques of Immunocytochemistry, Vol. 1 (edited byBullock, G. R. andPetrusz, P. pp. 1–75. New York, London: Academic Press.

    Google Scholar 

  • Bussolati, G. (1978) A fixation-decalcification procedure for bone biopsies.Histopathology 2, 329–34.

    Google Scholar 

  • Curran, R. C. &Gregory, J. (1978) Demonstration of immunoglobulin in cryostat and paraffin sections of human tonsil by immunofluorescence and immunoperoxidase techniques. Effects of processing on immunohistochemical performance of tissues and on the use of proteolytic enzymes to unmask antigens in sections.J. clin. Path. 31, 974–83.

    Google Scholar 

  • Curran, R. C. &Gregory, J. (1980) Effects of fixation and processing on immunohistochemical demonstration of immunoglobulin in paraffin sections of tonsil and bone marrow.J. clin. Path. 33, 1047–57.

    Google Scholar 

  • Heyderman, E. (1979) Immunoperoxidase technique in histopathology; application, methods and controls.J. clin. path. 32, 971–8.

    Google Scholar 

  • Jefferis, R., Mathews, J. B. &Bayley, P. M. (1978) Studies of human IgD myeloma proteins Conformational changes induced in the Fc fragment on heating or exposure to acid pH.Immunochemistry 15, 19–25.

    Google Scholar 

  • Leathem, A. &Atkins, N. (1980) Fixation and immunohistochemistry of lymphoid tissue.J. clin. Path. 33, 1010–2.

    Google Scholar 

  • Lillie, R. D. (1965)Histopathologic Technic and Practical Histochemistry, 3rd edn, p. 628. Maidenhead: McGraw-Hill.

    Google Scholar 

  • Matthews, J. B. (1981) Influence of clearing agent on immunohistochemical staining of paraffin-embedded tissue.J. clin. Path. 34, 103–5.

    Google Scholar 

  • Matthews, J. B. (1982) Influence of decalcification on immunohistochemical staining of formalin-fixed paraffin-embedded tissue.J. clin. Path. 35, 1392–4.

    Google Scholar 

  • Matthews, J. B. &Jefferis, R. (1977) The effects of chemical modification on the antigenicity of a human kappa Bence-Jones protein.Immunochemistry 14, 799–807.

    Google Scholar 

  • Matthews, J. B. &Basu, M. K. (1982) Plasma cell lesions within the oral tissues: Immunoperoxidase staining of routinely fixed and processed tissue.Oral Surg. 54, 414–23.

    Google Scholar 

  • Matthews, J. B. &Mason, G. I. (1983a) Granular cell myoblastoma: an immunoperoxidase study using a variety of antisera to human carcinoembryonic antigen.Histopathology 7, 77–82.

    Google Scholar 

  • Matthews, J. B. &Mason, G. I. (1983b) Immunoglobulin producing cells in human periapical granulomas.Br. J. Oral Surg. 21, 192–7.

    Google Scholar 

  • Mepham B. L. (1982) Influence of fixatives on the immunoreactivity of paraffin sections.Histochem. J. 14, 731–7.

    Google Scholar 

  • Mepham, B. L., Frater, W. &Mitchell, B. S. (1979) The use of proteolytic enzymes to improve immunoglobulin staining by the PAP technique.Histochem. J. 11, 345–58.

    Google Scholar 

  • Mukai, K. &Rosai, J. (1980) InProgress of Surgical Pathology, Vol. 1 (edited byFenoglio, C. M. andWolf, M.) pp. 15–49. New York: Masson Publishing USA, Inc.

    Google Scholar 

  • Robinson, G. (1982) InTheory and Practice of Histological Techniques, 2nd Edn. (edited byBancroft, J. D. andStevens, A.), Ch. 20. Edinburgh: Churchill Livingstone.

    Google Scholar 

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Matthews, J.B., Mason, G.I. Influence of decalcifying agents on immunoreactivity of formalin-fixed, paraffin-embedded tissue. Histochem J 16, 771–787 (1984). https://doi.org/10.1007/BF01095281

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  • DOI: https://doi.org/10.1007/BF01095281

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