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Double labelling of cell surface antigens with colloidal gold markers

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Summary

Particles of colloidal gold of different diameters (15nm and 40) have been used to distinctively label different surface antigens expressed on the surface of human peripheral blood B and T lymphocytes. Silver enhancement has been used to facilitate the observation of the gold particles. Observations were carried out in the backscattered electron imaging mode of the scanning electron microscope. Two different methods have been compared: in Method I the two antigens have been identified by monoclonal antibodies of different classes (IgG and IgM); in Method II monoclonal antibodies of the same subclass were used but the ligands were different (goat anti-murine Ig versus biotin/streptavidin). Some cross-reactivity was observed with Method I, but not with Method II.

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References

  • Bastholm, L., Nielsen, M. H. &Larson, L. I. (1987) Simultaneous demonstration of two antigens in ultrathin cryosections by a novel application of an immunogold staining method using primary antibodies from the same species.Histochemistry 87, 229–31.

    Article  PubMed  Google Scholar 

  • Bendayan, M. (1982) Double immunocytochemical labelling applying the protein A-gold technique.J. Histochem. Cytochem. 30, 81–5.

    PubMed  Google Scholar 

  • Bendayan, M. (1984) Protein A-gold electron microscopic immunocytochemistry: methods, application, and limitations.J. Electron Microsc. Tech. 1, 243–70.

    Article  Google Scholar 

  • Bienz, K., Egger, D. &Pasamontes, L. (1986) Electron microscopic immunocytochemistry. Silver enhancement of colloidal gold marker allows double labelling with the same primary antibody.J. Histochem. Cytochem. 34, 1337–42.

    PubMed  Google Scholar 

  • Danscher, G. &Norgaard, J. o. R. (1983) Light microscopic visualization of colloidal gold on resin-embedded tissue.J. Histochem. Cytochem. 31, 1394–8.

    PubMed  Google Scholar 

  • De Harven, E., Leung, R. &Christensen, H. (1984) A novel approach for scanning electron microscopy of colloidal gold-labelled cell surfaces.J. Cell Biol. 99, 53–7.

    PubMed  Google Scholar 

  • De Harven, E. &Soligo, D. (1986) Scanning electron microscopy of cell surface antigens labelled with colloidal gold.Am. J. Anat. 175, 277–87.

    Article  PubMed  Google Scholar 

  • De Harven, E., Soligo, D., Christensen, H. & Bhupal, V. (1986) Comparative surface morphology of human B-and T-derived peripheral lymphocytes. Proceedings of the 6th International Congress of Immunology, Toronto, p. 239.

  • De Harven, E. &Soligo, D. (1989) Backscattered electron imaging of the colloidal gold marker on cell surfaces. InColloidal Gold, Principles, Methods and Applications (edited byHayat, M. A.) San Diego: Academic Press, (in press).

    Google Scholar 

  • De Harven, E., Christensen, H., Leung, R. & Ackerley, C. (1989) Quantification of cell-surface expressed antigenic sites with 5nm gold markers: getting close to biologically relevant data.Proceedings of the 47th Annual Meeting of the Electron Microscopy Society of America. San Antonio: TX, p. 1050–1.

  • deMey, J. (1983) Colloidal gold probes in immunocytochemistry. InImmunocytochemistry: Practical Applications in Pathology and Biology (edited byPolak, J. & vanNoorden, S.) pp. 82–113. London: J. Wright.

    Google Scholar 

  • De Waele, M., De Mey, J., Renmans, W., Labeur, C., Jochmans, K. &Van Camp, B. (1986) Potential of immunogold-silver staining for the study of leukocyte subpopulations as defined by monoclonal antibodies.J. Histochem. Cytochem. 34, 1257–63.

    PubMed  Google Scholar 

  • Doerr-schott, J. &Lichte, C. M. (1986) A triple ultrastructural immunogold staining method. Application to the simultaneous demonstration of three hypophyseal hormones.J. Histochem. Cytochem. 34, 1101–4.

    PubMed  Google Scholar 

  • Goode, D. &Maugel, T. K. (1987) Backscattered electron imaging of immunogold-labelled and silver-enhanced microtubules in cultured mammalian cells.J. Electron Microsc. Tech. 5, 263–73.

    Article  Google Scholar 

  • Hoogeveen, Y. L., Smit, J. W., Blom, N. R. &Halie, R. M. (1988) Gold probe choice in simultaneous detection of human lymphocyte surface antigens at the ultrastructural level.J. Histochem. Cytochem. 36, 821–5.

    PubMed  Google Scholar 

  • Horisberger, M., Rosset, J. &Bauer, H. (1975) Colloidal gold granules as markers for cell surface receptors in the scanning electron microscope.Experimentia 31, 1147.

    Google Scholar 

  • Horisberger, M. &Rosset, J. (1977) Colloidal gold: a useful marker for transmission and scanning electron microscopy.J. Histochem. Cytochem. 25, 295–305.

    PubMed  Google Scholar 

  • Namork, E. &Heier, H. E. (1989) Silver enhancement of gold probes (5–40 nm): Single and double labelling of antigenic sites on cell surfaces imaged with backscattered electrons.J. Electron Microsc. Tech. 11, 102–8.

    Article  PubMed  Google Scholar 

  • Roth, J., Bendayan, M., &Orci, L. (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex.J. Histochem. Cytochem. 26, 1074–81.

    PubMed  Google Scholar 

  • Sako, H., Nakane, Y., Okino, K., Nishihara, K., Kodama, M., Kawata, M. &Yamada, H. (1986) Simultaneous detection of B-cells and T-cells by a double immunohistochemical technique using immunogold-silver staining and the avidin-biotin-peroxidase complex method.Histochemistry 86, 1–4.

    Article  PubMed  Google Scholar 

  • Scopsi, L., Larson, L. I., Bastholm, L. &Nielsen, M. H. (1986) Silver-enhanced colloidal gold probes as markers for scanning electron microscopy.Histochemistry 86, 35–41.

    Article  PubMed  Google Scholar 

  • Soligo, D., De Harven, E., Nava, M. T. &Lambertenghi-Deliliers, G. (1985) Immunocytochemistry with backscattered electrons. InScience of Biological Specimen Preparation (edited byMueller, M., Becker, R. P., Boyde, A. &Wolosewick, J. J.) pp. 289–297. Chicago: Scanning Electron Microscopy, Inc.

    Google Scholar 

  • Soligo, D., De Harven, E., Quirici, N. & Lambertenghi-Deliliers, G. (1987a) Studio immunologico delle sottopopolazioni linfocitarie al microscopio elettronico a scansione.Attualita in Ematologia, Genova, 947–50.

  • Soligo, D., Lambertenghi-Deliliers, G. &De Harven, E. (1987b) Immuno-scanning electron microscopy of normal and leukemic leukocytes labelled with colloidal gold.Scanning Microscopy 1, 719–25.

    PubMed  Google Scholar 

  • Walther, P. & Muller, M. (1985) Detection of small (5–15 nm) gold-labelled surface antigens by backscattered electrons.Proceedings of the 43rd Annual Meeting of the Electron Microscopy Society of America (edited by B-upailey, G. W.) pp. 538–41.

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De Harven, E., Soligo, D. & Christensen, H. Double labelling of cell surface antigens with colloidal gold markers. Histochem J 22, 18–23 (1990). https://doi.org/10.1007/BF01962875

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