1887

Abstract

Summary

Immunogold-silver staining is a sensitive staining technique that enables the visualisation of the presence of individual antigens by conventionallight microscopy. The application of this method to detect the antigenic heterogeneity of bacterial surface components and also the localisation of intracellular or extracellular bacteria is described. The latter applicationinvolved selective immuno-silver staining of the extracellular bacteria andcounterstaining of the intracellular bacteria and the eukaryotic cells bycrystal violet. The efficacy of the assay was confirmed by transmission electronmicroscopy of the silver-stained specimens. Immunogold-silver staining was shown to be useful for studying bacterial antigen variation and the uptake of bacteria by eukaryotic cells.

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1990-09-01
2024-04-23
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References

  1. Danscher G. Histochemical demonstration of heavy metals. A revised version of the sulphide silver method suitable for both light and electron microscopy. Histochemistry 1981; 71:1–16
    [Google Scholar]
  2. De Waele M, De Mey J, Renmans W, Labeur C, Reynaert P, van Camp B. Immunogold-silver staining of lymphocyte surface antigens on cells in suspension and in lymphnode cryostat sections. J Microsc 1986; 143:151–160
    [Google Scholar]
  3. Holgate CS, Jackson P, Cowen PN, Bird CC. Immunogold-silver staining: a new method of immunostaining with enhanced sensitivity. J Histochem Cytochem 1983; 31:938–944
    [Google Scholar]
  4. Springall DR, Hacker GW, Grimelius L, Polack JM. The potential of immunogold-silver staining method for paraffin sections. Histochemistry 1984; 81:603–608
    [Google Scholar]
  5. Brada D, Roth J. “Golden blot”—detection of polyclonal and monoclonal antibodies bound to antigens on nitrocellulose by protein A-gold complexes. Anal Biochem 1984; 142:79–83
    [Google Scholar]
  6. Tjia KF, Van Putten JPM, Pels E, Zanen HC. The interaction between Neisseria gonorrhoeae and the human cornea in organ culture. An electron microscopic study. Graefes Arch Clin Exp Ophthalmol 1988; 226:341–345
    [Google Scholar]
  7. Slot J W, Geuze HJ. A new method of preparing gold probes for multiple-labelling cytochemistry. Eur J Cell Biol 1985; 38:87–93
    [Google Scholar]
  8. Tam MR, Buchanan TM, Sandstrom EG et al. Serological classification of Neisseria gonorrhoeae with monoclonal antibodies. Infect Immun 1982; 36:1042–1053
    [Google Scholar]
  9. Poolman JT, Buchanan TM. Monoclonal antibody activity against native and denatured forms of gonococcal outer membrane proteins as detected by ultrathin, longitudinal slices of polyacrylamide gels. J Immunol Methods 1984; 75:265–274
    [Google Scholar]
  10. Weel JFL, Van Putten JPM. Ultrastructural localization of gonococcal antigens in infected epithelial cells as visualized by post-embedding immuno-electron-mi- croscopy. Microb Pathog 1988; 4:213–222
    [Google Scholar]
  11. Apicella MA, Shero M, Jarvis GA, Griffiss JM, Mandrell RE, Schneider H. Phenotypic variation of antigen expression of the lipooligosaccharide of Neisseria gonorrhoeae. In: Poolman JT, Zanen HC, Meyer TF et al. (eds) Gonococci and meningococci Dordrecht: Kluwer Academic Publishers; 1988477–483
    [Google Scholar]
  12. Meyer TF, Frosch M, Gibbs CP et al. Virulence functions and antigen variation in pathogenic Neisseriae. Antonie van Leeuwenhoek 1988; 54:421–430
    [Google Scholar]
  13. Meyer TF, Van Putten JPM. Genetic mechanisms and biological implications of phase variation in pathogenic neisseriae. Clin Microbiol Rev 1989; 2:S139–S145
    [Google Scholar]
  14. Apicella MA, Shero M, Jarvis GA, Griffiss JM, Mandrell RE, Schneider H. Phenotypic variation in epitope expression of the Neisseria gonorrhoeae lipooligosac-charide. InfectImmun 1987; 55:1755–1761
    [Google Scholar]
  15. Thomas DW, Hill JC, Tyeryar FJ. Interaction of gonococci with phagocytic leukocytes from men and mice. Infect Immun 1973; 8:98–104
    [Google Scholar]
  16. Finlay BB, Falkow S. Comparison of the invasion strategies used by Salmonella cholerae-suis, Shigella flexneri and Yersinia enterocolitica to enter cultured animal cells: endosome acidification is not required for bacterial invasion or intracellular replication. Biochimie 1988; 70:1089–1099
    [Google Scholar]
  17. McCoubrey WK, Howard LV. Incorporation of [14C]-methionine by Yersinia enterocolitica after invasion of HeLa cells. Infect Immun 1981; 32:956–959
    [Google Scholar]
  18. Hed J. The extinction of fluorescence by crystal violet and its use to differentiate between attached and ingested microorganisms in phagocytosis. FEMS Microbiol Lett 1977; 1:357–361
    [Google Scholar]
  19. Kihlstrom E. Infection of HeLa cells with Salmonella typhimurium 395 MS and MR 10 bacteria. Infect Immun 1977; 17:290–295
    [Google Scholar]
  20. Bukholm G, Johansen BV, Namork E, Lassen J. Bacterial adhesiveness and invasiveness in cell culture monolayer. Acta Pathol Microbiol Immunol Scand Sect B 1982; 90:403–408
    [Google Scholar]
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