Controlled growth of colloidal gold particles and implications for labelling efficiency.
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 beta-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.
D003102:Colloids; D004926:Escherichia coli / Q000201:enzymology* / Q000648:ultrastructure; D005696:Galactosidases / Q000032:analysis*; D006046:Gold; D008854:Microscopy, Electron / Q000379:methods; D013205:Staphylococcal Protein A; D001616:beta-Galactosidase / Q000032:analysis*
D003102:Colloids; D013205:Staphylococcal Protein A; D006046:Gold; D005696:Galactosidases; D001616:beta-Galactosidase