The Histochemical Journal

, Volume 14, Issue 5, pp 791–801 | Cite as

The preparation of protein A-gold complexes with 3 nm and 15 nm gold particles and their use in labelling multiple antigens on ultra-thin sections

  • Jurgen Roth
Papers

Summary

The preparation of a protein A-gold complex (pAg3) using 3 nm gold particles and its application for labelling of intracellular antigens on thin sections is reported. The 3 nm gold particle is the smallest metal particle currently available for cytochemistry and permits a higher resolution of the pAg technique. Furthermore, it can be used in double labelling experiments in conjunction with a pAg complex prepared from 15 nm gold particles. For double labelling, the pAg3 complex must be used for staining of the first antigen since otherwise a non-specific co-labelling of the two pAg complexes results.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Batten, T. F. C. &Hopkins, C. R. (1979) Use of protein A-coated colloidal gold particles for immunoelectronmicroscopic localization of ACTH on ultra-thin sections.Histochemistry 60, 317–20.Google Scholar
  2. Bendayan, M. (1980) Use of the protein A-gold technique for the morphological study of vascular permeability.J. Histochem. Cytochem. 28, 1251–4.Google Scholar
  3. Bendayan, M. (1981) Ultrastructural localization of actin in insulin-containing granules.Biol. Cell. 41, 157–60.Google Scholar
  4. Bendayan, M.,&Ørstavik, T. B. (1982) Immunocytochemical localization of kallikrein in the rat exocrine pancreas.J. Histochem. Cytochem. 30, 139–47.Google Scholar
  5. Bendayan, M., Roth, J., Perrelet, A. &Orci, L. (1980) Quantitative immunocytochemical localization of pancreatic secretory proteins in subcellular compartments of the rat acinar cell.J. Histochem. Cytochem. 28, 149–60.Google Scholar
  6. Bendayan, M. & Shore, G. C. (1982) Immunocytochemical localization of mitochondrial proteins in the rat hepatocyte.J. Histochem. Cytochem. (in press).Google Scholar
  7. Endresen, C. (1979) The binding to protein A of immunoglobulin G and of Fab and Fc fragments.Acta path. microbiol. scand. Sect. C 87, 185–9.Google Scholar
  8. Faulk, W. P. &Taylor, G. M. (1971) An immunocolloid method for the electron microscope.Immunochemistry 8, 1081–3.Google Scholar
  9. Forsgren, A. &Sjöquist, J. (1966) ‘Protein A’ fromS. aureus. I. Pseudoimmune reaction with human γ-globulin.J. Immun. 97, 822–7.Google Scholar
  10. Forsgren, A. &Sjöquist, J. (1967) ‘Protein A’ fromS. aureus. III. Reaction with rabbit γ-globulin.J. Immun. 99, 19.Google Scholar
  11. Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold solutions.Nature Phys. Sci. 241, 20–2.Google Scholar
  12. Grov, A. &Endresen, C. (1976) An examination of the ‘star phenomenon’, a three component immunoprecipitation involving staphylococcal protein A.Acta path. microbiol. scand. Sect. C 84, 333–6.Google Scholar
  13. Geuze, J. J., Slot, J. W. &Brands, R. (1981b) The occurrence of albumin in the rat liver. A light and electron microscope immunocytochemical study.Cell Biol. Int. Rep. 5, 463.Google Scholar
  14. Geuze, H. J., Slot, J. W., Van Der Ley, P. A. &Scheffer, R. C. T. (1981b) Use of colloidal gold particles in double-labelling immunoelectron microscopy of ultrathin frozen tissue sections.J. Cell Biol. 89, 653–65.Google Scholar
  15. Green, J., Griffiths, G., Louvard, D., Quinn, P. &Warren, G. (1981) Passage of viral membrane proteins through the Golgi complex.J. molec. Biol. 152, 663–98.Google Scholar
  16. Horisberger, M. (1979) Evaluation of colloidal gold as a cytochemical marker for transmission and scanning electron microscopy.Biol. Cell. 36, 253–8.Google Scholar
  17. Horisberger, M. &Rosset, J. (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy.J. Histochem. Cytochem. 25, 295–305.Google Scholar
  18. Horisberger, M. &Von Lanthen, M. (1978) Simultaneous localization of a hepatic binding protein specific for galactose and of galactose-containing receptors on rat hepatocytes.J. Histochem. Cytochem. 26, 960–6.Google Scholar
  19. Nakane, P. K. (1968) Simultaneous localization of multiple tissue antigens using the peroxidase-labelled antibody method. A study on pituitary glands of the rat.J. Histochem. Cytochem. 16, 557–60.Google Scholar
  20. Nygren, H., Rozell, B., Holmgren, A. &Hansson, H.-A. (1981) Immunoelectron microscopic localization of glutaredoxin and thioredoxin inEscherichia coli cells.FEBS Lett. 133, 145–50.Google Scholar
  21. Ravazzola, M. &Orci, L. (1980) Glucagon and glicentin immunoreactivity are topologically segregated in the α-granule of human pancreatic A cell.Nature, Lond. 284, 66–67.Google Scholar
  22. Romano, E. L. &Romano, M. (1977) Staphylococcal protein A bound to colloidal gold: a useful reagent to label antigen-antibody sites in electron microscopy.Immunochemistry 14, 711–5.Google Scholar
  23. Roth, J. (1982a) The protein A-gold (pAg) technique. A qualitative and quantitative approach for antigen localization on thin sections. InTechniques in Immunocytochemistry (edited byBullock, G. B. andPetrusz, P.) New York, London: Academic Press.Google Scholar
  24. Roth, J. (1982b) Applications of immunocolloids in light microscopy. Preparation of protein A-silver and protein A-gold complexes and their application for localization of single and multiple antigens in paraffin sections.J. Histochem. Cytochem. (in press).Google Scholar
  25. Roth, J. &Binder, M. (1978) Colloidal gold, ferritin and peroxidase as markers for electron microscopic double labelling lectin techniques.J. Histochem. Cytochem. 26, 163–9.Google Scholar
  26. Roth, J. &Berger, E. C. (1982) Immunocytochemical localization of galactosyltransferase in HeLa cells: codistribution with thiamine pyrophosphatase in trans Golgi cisternae.J. Cell Biol. 93, 223–9.Google Scholar
  27. 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.Google Scholar
  28. Roth, J., Bendayan, M. &Orci, L. (1980) FITC-protein A-gold complex for light and electron microscopic immunocytochemistry.J. Histochem. Cytochem. 28, 55–7.Google Scholar
  29. Roth, J., Ravazzola, M., Bendayan, M. &Orci, L. (1981a) Application of the protein A-gold technique for electron microscopic demonstration of polypeptide hormones.Endocrinology 108, 247–53.Google Scholar
  30. Roth, J., Thorens, B., Hunziker, W., Norman, A. W. &Orci, L. (1981b) Vitamin D-dependent calcium binding protein: immuncytochemical localization in chick kidney.Science 214, 197–200.Google Scholar
  31. Roth, J., Bendayan, M., Carlemalm, E., Villiger, W. &Garavito, M. (1981c) Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue.J. Histochem. Cytochem. 29, 663–71.Google Scholar
  32. Slot, J. W. &Geuze, J. J. (1981) Sizing of protein A-colloidal gold probes for immunoelectronmicroscopy.J. Cell Biol. 90, 533–6.Google Scholar
  33. Tanaka, T., Gresik, E. W., Michelakis, A. M. &Barka, T. (1980) Immunocytochemical localization of renin in kidneys and submandibular glands of SWR/J and C 57 Bl/6J mice.J. Histochem. Cytochem. 28, 1113–8.Google Scholar
  34. Tanaka, T., Gresik, E. W. &Barka, T. (1981a) Immunocytochemical localization of amylase in the parotid gland of developing and adult rats.J. Histochem. Cytochem. 29, 1189–95.Google Scholar
  35. Tanaka, T., Gresik, E. W. &Barka, T. (1981b) Epidermal growth factor and renin in mouse submandibular glands.J. Histochem. Cytochem. 29, 1229–31.Google Scholar
  36. Thorens, B., Roth, J., Norman, A. W., Perrelet, A. & Orci, L. (1982) Immunocytochemical localization of the vitamin D-dependent calcium binding protein in chick duodenum.J. Cell. Biol. (in press).Google Scholar
  37. Zikan, J. (1980) Interactions of pig Fab γ fragments with protein A fromStaphylococcus aureus.Folia Microbiol. 25, 246–53.Google Scholar
  38. Zsigmondy, R. (1905)Zur Erkenntnis der Kolloide. Jena.Google Scholar
  39. Zsigmondy, R. &Thiessen, P. A. (1925)Das Kolloidale Gold. Leipzig: Akademische Verlagsgesellschaft.Google Scholar

Copyright information

© Chapman and Hall Ltd. 1982

Authors and Affiliations

  • Jurgen Roth
    • 1
  1. 1.Institute of Histology and EmbryologyUniversity of Geneva Medical SchoolGeneva 4Switzerland

Personalised recommendations