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Light and electron microscopical combined staining by immunohistochemical and enzyme histochemical methods using rat prolactin-secreting pituitary tumours

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Summary

Combined immunohistochemical staining (IHCS) and enzyme histochemical staining (EHCS) methods for light microscopy (LM) and electron microscopy (EM) are reported, using oestrogeninduced rat pituitary tumours. For LM, combined staining for alkaline phosphatase and acid phosphatase by EHCS, using the azo dye method, and for prolactin and ACTH by IHCS, using the enzyme-labelled antibody method, gave the best results on 1 μm glycol methacrylate sections. For EM, combined staining by EHCS on 30 μm tissue sections followed by IHCS for prolactin on ultrathin Epon sections (enzyme-labelled antibody method) provided acceptable results. By these combined staining methods, the neoplastic prolactin cells were shown to have close affinity to rich alkaline phosphatase-positive capillaries and to possess an alkaline phosphatase-positive cell membrane. Furthermore, they revealed acid phosphatase-positive lysosomal and secretory granules. These combined staining methods may be valuable in studies on the actual functional status of cells.

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References

  • Ashford, A. E., Allaway, U. G. &McCully, M. E. (1972) Low temperature embedding in glycol methacrylate for enzyme histochemistry in plant and animal tissues.J. Histochem. Cytochem. 20, 986–90.

    Google Scholar 

  • Barka, T. &Anderson, P. F. (1962) Histochemical methods for acid phosphatase using hexazonium pararosanilin as coupler.J. Histochem. Cytochem. 10, 741.

    Google Scholar 

  • Hoshino, M. (1971) The use of glycol methacrylate as an embedding medium for the histochemical demonstration of acid phosphatase activity.J. Histochem. Cytochem. 19, 575–7.

    Google Scholar 

  • Hoshino, M. &Kobayashi, H. (1973) Glycol methacrylate embedding in immunocytochemical methods.J. Histochem. Cytochem. 20, 743–5.

    Google Scholar 

  • Ito, A. (1976) Pituitary tumours in rats (animal model of human disease).Am. J. Pathol. 83, 423–6.

    Google Scholar 

  • Kawarai, Y. &Nakane, P. K. (1970) Localization of tissue antigens on the ultrathin sections with peroxidase-labeled antibody method.J. Histochem. Cytochem. 18, 161–6.

    Google Scholar 

  • Mayahara, H., Hirano, H., Saito, T. &Ogawa, K. (1967) The new lead citrate method for the ultracytochemical demonstration of activity of non-specific alkaline phosphatase (orthophosphoric monoester phosphohydrolase).Histochemie 11, 88–96.

    Google Scholar 

  • Nakane, P. K. (1971) Application of peroxidase-labeled antibodies to the intracellular localization of hormones.Acta endocrinol. suppl.153, 190–204.

    Google Scholar 

  • Nakane, P. K. (1975) Recent progress in peroxidase-labeled antibody method.Ann. N.Y. Acad. Sci. 254, 203–11.

    Google Scholar 

  • Nakane, P. K. &Kawaoi, A. (1974) Peroxidase-labeled antibody. A new method of conjugation.J. Histochem. Cytochem. 22, 1084–91.

    Google Scholar 

  • Novikoff, A. B. (1963). Lysosomes in the physiology and pathology of cells: contributions of staining methods. InCiba Foundation Symposium: Lysosomes (edited byde Reuck, A. V. S. andCameron, M. P.), pp. 36–73. Churchill: London.

    Google Scholar 

  • Osamura, R. Y. &Watanabe, K. (1976) Peroxidase-labeled antibody method. Its principles and applications to endocrinology.Clin. Endocrinol. 24, 1074–87 (in Japanese).

    Google Scholar 

  • Osamura, R. Y., Akatsuka, A. &Watanabe, K. (1978a) Localization of anterior pituitary hormones on Epon sections by peroxidase-labeled antibody method-Light and electron-microscopic observations.Acta histochem. cytochem. 11, 399–407.

    Google Scholar 

  • Osamura, R. Y., Watanabe, K., Teramoto, A., Hirakawa, K., Kawano, N. &Morii, S. (1978b) Male prolactin secreting pituitary adenomas in human studied by peroxidase-labeled antibody method.Acta endocrinol. 88, 643–52.

    Google Scholar 

  • Spaur R. C. &Moriarty, G. C. (1977) Improvement of glycol methacrylate. 1. Its use as an embedding medium for electron microscopic studies.J. Histochem. Cytochem. 25, 163–74.

    Google Scholar 

  • Ueda, G., Moy, P. &Furth, J. (1973) Multihormonal activities of normal and neoplastic pituitary cells as indicated by immunohistochemical staining.Int. J. Cancer 12, 100–14.

    Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, Tokai University, School of Medicine, Boseidai, 259-11, Isehara City, Kanagawa, Japan

    R. Yoshiyuki Osamura & Etsuko Nakahashi

  2. Cell Biology Research Laboratory, Tokai University, School of Medicine, Boseidai, 259-11, Isehara-City, Kanagawa, Japan

    Noriyuki komatsu

Authors
  1. R. Yoshiyuki Osamura
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  2. Noriyuki komatsu
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  3. Etsuko Nakahashi
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  4. Keiichi Watanabe
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Osamura, R.Y., komatsu, N., Nakahashi, E. et al. Light and electron microscopical combined staining by immunohistochemical and enzyme histochemical methods using rat prolactin-secreting pituitary tumours. Histochem J 12, 371–379 (1980). https://doi.org/10.1007/BF01011955

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  • DOI: https://doi.org/10.1007/BF01011955

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