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Investigation of immunogold-silver staining by electron microscopy

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Summary

Deposition of metallic silver on colloidal gold immunoreagents has been shown to be a very sensitive immunostaining technique capable of detecting low levels of immunoreactivity in tissue sections. Using electron microscopy we have shown that immunolabelling is highest with small sizes of gold which can penetrate sections better and achieve higher densities of particles in the section than larger particles. Chemical permeabilisation of the embedding medium aids the penetration of colloidal gold. The silver enhancement step in immunogold-silver staining was shown to be progressive, allowing optimisation of staining and the selection of the final size of silver deposits required. Some poorly understood features of the technique are rationalised and the additional knowledge gained will aid the wider application of this method.

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References

  • Arens H, Eggert J (1929) Das Wachstum des kolloiden Silbers in Gelatineschichten. Z Elektrochem 35:728–733

    Google Scholar 

  • Danscher G (1981a) Localisation of gold in biological tissue. Histochemistry 71:81–88

    Google Scholar 

  • Danscher G (1981b) Light and electron microscopic localisation of silver in biological tissue. Histochemistry 71:177–186

    Google Scholar 

  • Danscher G (1981c) Histochemical demonstration of heavy metals. Histochemistry 71:1–16

    Google Scholar 

  • Danscher G (1982) Exogenous selenium in the brain. Histochemistry 76:281–293

    Google Scholar 

  • Danscher G, Nörgaard JOR (1983) Light microscopic visualisation of colloidal gold on resin embedded tissue. J Histochem Cytochem 31:1394–1398

    Google Scholar 

  • Gu J, De Mey J, Moeremans M, Polak JM (1981) Sequential use of the PAP and immunogold staining methods for the light microscopical double staining of tissue antigens: its application to the study of regulatory peptides in the gut. Regul Pept 1:365–374

    Google Scholar 

  • Faulk WP, Taylor GM (1971) An immunocolloid method for the electron microscope. Immunochemistry 8:1081–1083

    Google Scholar 

  • Hacker GW, Springall DR, Van Noorden S, Bishop AE, Grimelius L, Polak JM (1985) The immunogold staining method — a powerful tool in histopathology. Virchows Arch (A) 406:449–461

    Google Scholar 

  • Holgate CS, Jackson P, Cowen PN, Bird CC (1983a) Immunogoldsilver staining: new method of immunostaining with enhanced sensitivity. J Histochem Cytochem 31:938–944

    Google Scholar 

  • Holgate CS, Jackson P, Lauder I, Cowen PN, Bird CC (1983b) Surface membrane staining of immunoglobins in paraffin sections of non-Hodgkin's lymphomas using the immunogold silver staining technique. J Clin Pathol 36:742–746

    Google Scholar 

  • Horisberger M (1981) Colloidal gold: a cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. Scan Electron Microsc II:9–31

    Google Scholar 

  • Huang WM, Gibson SJ, Facer P, Gu J, Polak JM (1983) Improved section adhesion for immunocytochemistry using high molecular weight polymers of l-lysine as a slide coating. Histochemistry 77:275–279

    Google Scholar 

  • James TH (1939) Reduction of silver ions by hydroquinone. J Am Chem Soc 61:648–658

    Google Scholar 

  • James TH (1977) The theory of the photographic process. Macmillan, New York

    Google Scholar 

  • Moeremans M, Daneels G, Van Dijck A, Langanger G, De Mey J (1984) Sensitive visualization of antigen-antibody reactions in dot and blot immuneoverlay assays with immunogold and immunogold/silver staining. J Immunol Methods 74:352–360

    Google Scholar 

  • Polak JM, Varndell IM (1984) Immunolabelling for electron microscopy. Elsevier, Amsterdam

    Google Scholar 

  • Scheuer PJ, Thorpe MEC, Marriott P (1967) A method for the demonstration of copper under the electron microscope. J Histochem Cytochem 15:300–301

    Google Scholar 

  • Springall DR, Hacker GW, Grimelius L, Polak JM (1984) The potential of the immunogold-silver staining method for paraffin sections. Histochemistry 81:603–608

    Google Scholar 

  • Varndell I, Tapia FJ, Probert L, Buchan AMJ, Gu J, De Mey J, Bloom SR, Polak JM (1982) Immunogold staining procedure for the localisation of regulatory peptides. Peptides 3:259–272

    Google Scholar 

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Authors and Affiliations

  1. Histochemistry Department, Royal Postgraduate Medical School, Hammersmith Hospital, Du Cane Road, W12 OHS, London, UK

    P. M. Lackie, R. J. Hennessy, G. W. Hacker & J. M. Polak

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  1. P. M. Lackie
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  2. R. J. Hennessy
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  3. G. W. Hacker
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  4. J. M. Polak
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Lackie, P.M., Hennessy, R.J., Hacker, G.W. et al. Investigation of immunogold-silver staining by electron microscopy. Histochemistry 83, 545–550 (1985). https://doi.org/10.1007/BF00492458

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  • DOI: https://doi.org/10.1007/BF00492458

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