Abstract
Electron microscopy permits the detailed study of cell relationships within tissues and organelles within cells. Two ultrastructural techniques in which gold probes have proven invaluable are immunocytochemistry and in situ hybridization, and these will be described here. Until 1980, peroxidase was the marker of choice, but now colloidal gold is almost universally used. The advantages of colloidal gold are that:
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1.
It is particulate and very dense, therefore easily identified on biological sections;
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2.
It is small and will not obscure fine structural details,
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3.
It can be prepared in several different sizes for use in multiple labeling experiments and quantitation, and
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4.
It is easily conjugated to immunoglobulins or proteins A or G.
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Beesley, J. (1989) Colloidal gold: a new perspective for cytochemical marking. Royal Microscopical Society Handbook 17. Oxford Science Publications, Oxford, UK.
Gross, U., Bormuth, H., Gaissmaier, C, Dittrich, C, Krenn, V., Bohne, W, and Ferguson, D. J. P. (1995) Monoclonal rat antibodies directed against Toxoplasma gondii suitable for studying tackyzoite-bradyzoite interconversion in vivo Clin Diag Lab Immunol 2, 542–548
Levy, E R. and Hemngton, C. S (eds). (1995) Nonisotopic Methods in Molecular Biology—A Practical Approach Oxford University Press, Oxford
Morel, G. (ed). (1993) Hybridization Techniques for Electron Microscopy CRC, Boca Raton, FL
Morey, A L, Ferguson, D J P, Leslie, K O., Taatses, D. J, and Fleming, K. A (1993) Detection of parvoviral and human DNA sequences at the ultrastructural level by in situ hybridization with digoxigenin-labeled probes. Histochem J 25, 421–429
Morey, A L. (1995) Nonisotopic in situ hybridization at the ultrastructural level J Pathol 176, 113–121
Morey, A L., Ferguson, D. J P., and Fleming, K. A. (1995) Combined immunocytochemistry and nonisotopic in situ hybridization for ultrastructural investigation of human parvovirus B19 infection Histochem J 27, 46–53.
Slot, J W. and Geuze, H J (1984) Gold markers for single and double immunolabeling of ultrathin cryosections, in Immunolabehng for Electron Microscopy (Polak, J M. and Varndell, I. M, eds), Elsevier, Amsterdam, pp 129–142
Tokuyasu, K T. (1978) A study of positive staining of ultrathin frozen sections. J Ultrastruct Res 63, 287–307
Reynolds, E. S. (1963) The use of lead citrate at high pH as an electron-opaque stain in electron microscopy J Cell Biol 17, 208–212
Carlemalm, E, Garavito, R M., and Villiger, W (1982) Resin development for electron microscopy and an analysis of embedding at low temperature J Microsc 126, 123–143.
Griffiths, G, McDowall, A., Back, R, and Dubochet, J (1984) On the preparation of cryosections for immunocytochemistry J Ultrastruct Res 89, 68–78
Tokuyasu, K. T. (1986) Immunocryoultramicrotomy J Microsc 143, 139–149
Mannweiler, K., Hohenberg, H., Bohn, W., and Rutter, G. (1982) Protein A-gold particles as markers in replica immunocytochemistry. high resolution electron microscope investigations of plasma membrane surfaces J Microsc 126, 145–149
Hodges, G M., Southgate, J, and Toulson, E. C (1987) Colloidal gold-a powerful tool in S E. M. immunocytochemistry an overview of bioapplications. Scanning Electron Microscopy 1, 301–318.
Murphy, J A. (1980) Noncoating techniques to render biological specimens conductive/1980 update Scanning Electron Microscopy 1, 209–220
mnPinto da Silva, P., Barbosa, M. L. F, and Aguas, A P. (1986) A guide to fracture label, cytochemical labeling of freeze-fractured cells, in Advanced Techniques in Biological Electron Microscopy (Koehler, J K, ed), Springer-Verlag, Berlin, pp 201–227
mnPinto da Silva, P. and Kan, F. W. (1984) Label-fracture a method for high resolution labeling of cell surfaces J Cell Biol 99, 1156–1161
Robards, A. W. and Sleytr, U. B (1985) Low temperature methods in biological electron microscopy, in Practical Methods in Electron Microscopy, vol 10 (Glauert, A M, ed), Elsevier, Amsterdam
Robertson, D, Monaghan, P., Clark, C, and Atherton, A. J (1992) An appraisal of low temperature embedding by progressive lowering of temperature into Lowicryl HM20 for immunocytochemical studies J Microsc. 168, 85–100
Dudek, R W, Varndell, I. M., and Polak, J. M (1984) Combined quick-freeze and freeze-drying techniques for improved electron immunocytochemistry, in Immunolabeling for Electron Microscopy (Polak, J M and Varndell, I M, eds.), Elsevier, Amsterdam, pp 235–248
Oprins, A D., Geuze, H J, and Slot, J W (1994) Cryosubstitution dehydration of aldehyde-fixed tissue, a favourable approach to quantitative immunocytochemistry J Histochem Cytochem 42, 497–503
Monaghan, P and Robertson, D (1990) Freeze-substitution without aldehyde or osmium fixatives ultrastructural and implication for immunocytochemistry. J Microsc 158, 355–363
Shimizu, H. and Nishikawa, T. (1993) Application of an image analyser to gold labeling in immunoelectron microscopy to achieve better demonstration and quantitative analysis J Histochem Cytochem 41, 123–128
Morris, R. E, Ciraolo, G M, and Saelinger, C. B (1991) Gold enhancement of gold-labeled probes gold-intensified staining technique (GIST). J Histochem Cytochem 39, 1585–1591.
Lackie, P M, Hennessy, R J, Hacker, G W, and Polak, J M (1985) Investigation of immunogold-silver staining using electron microscopy Histochemistry 83, 545–550
Manara, G C, Ferrari, C, Torresam, C, Sansoni, P, and De Panfilis, G. (1990) The immunogold-silver staining approach in the study of lymphocyte subpopulations in transmission electron microscopy. J Immunol Methods 128, 59–63
Otsuki, Y, Maxwell, L. E, Magan, S., and Kubo, H (1990) Immunogold-silver staining method for light and electron microscopic detection of lymphocyte cell surface antigens with monoclonal antibodies J Histochem Cytochem 38, 1215–1221.
Burry, R. W., Vandrè, D D., and Hayes, D M. (1992) Silver enhancement of gold antibody probes in preembedding electron microscopic immunocytochemistry J Histochem Cytochem 40, 1849–1856
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© 1998 Humana Press Inc., Totowa, NJ
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Ferguson, D.J.P., Hughes, D.A., Beesley, J.E. (1998). Immunogold Probes in Electron Microscopy. In: Pound, J.D. (eds) Immunochemical Protocols. Methods in Molecular Biology™, vol 80. Humana Press. https://doi.org/10.1007/978-1-59259-257-9_31
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DOI: https://doi.org/10.1007/978-1-59259-257-9_31
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