Summary
A new method is reported for the preparation of colloidal gold particles with diameters ranging between 5 and 12 nm. The initial gold particle population, with an average diameter of 5.6±0.9 nm, is prepared by reduction of chloroauric acid with white phosphorous. An increase in particle diameter by growth is obtained by reduction of chloroauric acid with white phosphorous in the presence of colloidal gold particles. The labelling efficiency of these gold particles, conjugated with protein A, in indirect immunolabelling experiments is investigated by labelling of β-galactosidase on ultrathin cryosections of Escherichia coli cells. We demonstrate that the labelling efficiency is at least dependent on particle diameter, probe concentration and preparation method. In addition it is shown, that with this new method, gold particle populations can be prepared with minor overlap in diameter spreading. Therefore these gold probes are suitable for qualitative double labelling experiments. The quantitative aspect of immunolabelling is discussed.
Similar content being viewed by others
References
Brada D, Roth J (1984) ‘Golden blot’-Detection of polyclonal and monoclonal antibodies bound to antigens on nitrocellulose by protein A-gold complexes. Anal Biochem 142:79–83
De Mey J (1983) Colloidal gold probes in immunochemistry. In: Polak S, Van Noorden S (eds) Immunochemistry. Praotical applications in pathology and biology. Wright-PSG, London, pp 82–112
De Mey J, Moeremans M, Geuens G, Nuydens R, De Brabander M (1981) High resolution light and electron microscopic localization of tubulin with the IGS (immuno gold staining) method. Cell Biol Int Rep 5:889–899
Faulk W, Taylor G (1971) An immunocolloid method for the electron microscope. Immunochemistry 8:1081–1083
Geoghegan W, Ackerman G (1977) Absorbtion of horseradish peroxidase, ovomucoid and anti-immunoglobulin to colloidal gold for the indirect detection of concanavaline A, wheat germ agglutinin and goat anti-human immunoglobulin G on cell surfaces at the electron microscopic level: a new method, theory and application. J Histochem Cytochem 25:1187–1200
Geuze HJ, Slot JW, Van der Ley P, Schuffer R, Griffith J (1981) Use of colloidal gold particles in double labelling immunoelectron microscopy on ultrathin frozen sections. J cell Biol 89:653–665
Griffiths G, Brands R, Burke B, Louvard D, Warren G (1982) Viral membrane proteins acquire galactose in trans golgi cisternae during intracellular transport. J Cell Biol 95:781–792
Holgate C, Jackson P, Cowen P, Bird C (1983) Immunogold-silver staining: new method of immunostaining with enhanced sensitivity. J Histochem Cytochem 31:938–944
Horisberger M (1981) Colloidal gold: a cytochemical marker for light and fluorescent microscopy and for transmission and scanning electron microscopy. Scan Electron Microsc 2:9–28
Horisberger M (1983) Colloidal gold as a tool in molecular biology. Trends Biochem Sci 7:395–397
Horisberger M, Rosset J (1977) Colloidal gold, a useful marker for transmission and scanning electron microscopy. J Histochem Cytochem 25:295–305
Horisberger M, Vonlanthen M (1979) Multiple marking of cell surface receptors by gold granules: simultaneous localization of three lectin receptors on human erythrocyte. J Microsc 115:97–102
Hutchison N, Langer-Safer P, Ward D, Hamkalo B (1982) In situ hybridization at the electron microscope level: hybrid detection by autoradiography and colloidal gold. J Cell Biol 95:609–618
Moeremans M, Daneels G, De Mey J (1985) Sensitive colloidal metal (gold or silver) staining of protein blots on nitrocellulose membranes. Anal Biochem 145:315–321
Romano EL, Romano M (1977) Staphylococcal protein A bound to colloidal gold: a useful reagent to label antigen-antibody sites in electronmicroscopy. Immunochemistry 14:711–715
Romano EL, Romano M (1984) Historical aspects. In: Polak JM, Varndell IM (eds) Immunolabelling for electronmicroscopy. Elsevier, Amsterdam, pp 3–15
Romano EL, Stolinsky C, Hughes-Jones NC (1974) An antiglobulin reagent labelled with colloidal gold for use in electronmicroscopy. Immunochemistry 11:521–522
Slot JW, Gueze HJ (1981) Sizing of protein A-colloidal gold probes for immunoelectron microscopy. J Cell Biol 90:533–536
Slot JW, Geuze HJ (1984) Gold markers for single and double immunolabelling of ultrathin cryosections. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elsevier, Amsterdam, pp 129–142
Slot JW, Geuze HJ (1985) A new method of preparing gold probes for multiple-labeling cytochemistry. Eur J Cell Biol 38:87–93
Surolia A, Pain D, Khan MI (1982) Protein A: nature's universal anti-antibody. Trend Biochem Sci 7:74–76
Tokuyasu KT (1980) Immunocytochemistry on ultrathin frozen sections. Histochem J 12:381–403
Tokuyasu KT (1983) Present state of immunocryoultramicrotomy. J Histochem Cytochem 31:164–167
Van der Ley P, Amesz H, Tommassen J, Lugtenberg B (1985) Monoclonal antibodies directed against the cell-surface-exposed part of PhoE pore protein of the Escherichia coli K12 outer membrane. Eur J Biochem 147:401–407
Vernooy-Gerritsen M, Leunissen JLM, Veldink GA, Vliegenthart JFG (1984) Intracellular localization of lipoxygenases-1 and-2 in germinating soybean seeds by indirect labelling with protein A-colloidal gold complexes. Plant Physiol 76:1070–1079
Voorhout WF, Leunissen-Bijvelt JJM, Leunissen JLM, Verkleij AJ (1985) Steric hindrance in immunolabelling. J Microsc (in press)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
van Bergen en Henegouwen, P.M.P., Leunissen, J.L.M. Controlled growth of colloidal gold particles and implications for labelling efficiency. Histochemistry 85, 81–87 (1986). https://doi.org/10.1007/BF00508657
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00508657