Summary
The suitability of three acrylic resins for the immunoelectron microscopical localization of cell surface and cytoskeletal antigens in surgically excised, immersion fixed human breast cancer, using an immunogold system, has been assessed.
Good localization of milk fat globule membrane was achieved with LR White, LR Gold and Lowicryl K11M, although the embedding schedule for LR White had to be modified. The best results were achieved with Lowicryl K11M. Only scanty labelling of actin and cytokeratin was seen in LR White embedded tissue, whereas there was clear localization in LR Gold and Lowicryl K11M embedded samples. Tubulin and α-actinin was detected at low level in tissues in the low temperature embedding resins, but not in LR White embedded samples. The morphology of the latter was poorer, and there was greater variability in ultrastructure and labelling.
Of the two low temperature embedding resins, Lowicryl K11M gave slightly better results. However, the advantages could be outweighed by the problem incurred in achieving the low temperatures, and by poorer handling properties than LR Gold.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Acetarin, J.-D., Carlemalm, E. &Villiger, W. (1986) Developments of new Lowicryl® resins for embedding biological specimens at even lower temperatures.J. Microsc. 143, 81–8.
Armbruster, B. L., Garavito, R. M. &Kellenberger, E. (1983) Dehydration and embedding temperatures affect the antigenic specificity of tubulin and immunolabelling of the Protein A-colloidal gold technique.J. Histochem. Cytochem. 31, 1380–4.
Bendayan, M. &Shore, G. C. (1982) Immunohistochemical localization of mitochondrial proteins in the rat hepatocyte.J. Histochem. Cytochem. 30, 139–47.
Bendayan, M. (1983) Ultrastructural localization of actin in muscle, epithelial and secretory cells by applying the protein A-gold immunocytochemical technique.Histochem. J. 15, 39–58.
Bendayan, M., Nanci, A. &Kan, F. W. K. (1987) Effect of tissue processing on colloidal gold cytochemical technique.J. Histochem. Cytochem. 35, 983–96.
Carlemalm, E., Garvito, R. M. &Villiger, W. (1982) Resin development for electron microscopy and an analysis of embedding at low temperature.J. Microsc. 126, 123–43.
Frens, G. (1973) Controlled nucleation for the regulation of particle size in monodisperse gold solutions.Nat. Phys. Sci. 241, 20–2.
Kellenberger, E., Durrenberger, M., Villiger, W., Carlemalm, E. &Wurtz, M. (1987) The efficiency of immunolabel on Lowicryl sections compared to theoretical predictions.J. Histochem. Cytochem. 35, 959–69.
Lemanski, L. F., Paulson, D. J., Hill, C. S., Davis, L. A., Riles, L. C. &Lim, S. S. (1985) Immunoelectron microscopic localization of α-actinin on Lowicryl-embedded thin-sectioned tissues.J. Histochem. Cytochem. 33, 515–22.
Loesser, K. E., Doane, K. J., Wilson, F. J., Roisen, F. J. &Malamed, S. (1986) Improved immunoelectron microscopic method for localizing cytoskeletal proteins in Lowicryl K4M embedded tissues.J. Histochem. Cytochem. 34, 1477–85.
Major, P., Lavallee, M., Minassian, H., Kovac, P. &Wang, N.-S. (1987) Ultrastructural localization of a breast tumour-associated antigen.J. Histochem. Cytochem. 35, 375–9.
Maldonado, C. A. &Aoki, A. (1986) Influence of embedding media in prolactin labelling with immunogold techniques.Histochem. J. 18, 429–33.
McPhail, G. D., Finn, T. &Isaacson, P. G. (1987) A useful low temperature method for post-embedding electron immunocytochemistry in routine histopathology.J. Pathol. 151, 231–8.
Newman, G. R., Jasani, B. &Williams, E. D. (1983) A simple post-embedding system for the rapid demonstration of tissue antigens under the electron microscope.Histochem. J. 15, 543–5.
Newman, G. R. &Jasani, B. (1984) Post embedding immunoenzyme techniques. InImmuno-labelling for Electron Microscopy (edited byPolak, J. M. &Varndell, I. M.) pp. 53–70. Amsterdam, New York: Elsevier.
Newman, G. R. &Hobot, J. A. (1987) Modern acrylics for post-embedding immunostaining techniques.J. Histochem. Cytochem. 34, 971–81.
Petersen, O. W. &Van Deurs, B. (1986) Characterization of epithelial membrane antigen expression in human mammary epithelium by ultra-structural immunoperoxidase cytochemistry.J. Histochem. Cytochem. 34, 801–9.
Roth, J. (1982) The protein A-gold (pAg) technique — a qualitative and quantitative approach for antigen localization on thin sections. InTechniques in Immunocytochemistry (edited byBullock, G. R. &Petrusz, P.) Vol. 1, pp. 107–33. London: Academic Press.
Roth, J. (1983) The colloidal gold marker system for light and electron microscopic cytochemistry. InTechniques in Immunocytochemistry (edited byBullock, G. R. &Petrusz, P.) Vol. 2, pp. 217–84. London: Academic Press.
Roth, J. (1986) Post-embedding cytochemistry with goldlabelled reagents: a review.J. Microsc. 143, 125–37.
Roth, J., Bendayan, M., Carlemalm, E., Villiger, W. &Garvito, M. (1981) Enhancement of structural preservation and immunocytochemical staining in low temperature-embedded pancreatic tissue.J. Histochem. Cytochem. 29, 663–71.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Corcoran, D., Walker, R.A. Post embedding immunoelectron microscopy of human breast cancer: a comparison of three acrylic resins. Histochem J 22, 291–298 (1990). https://doi.org/10.1007/BF01387184
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01387184