van Bergen en Henegouwen P M, Leunissen J L
Histochemistry. 1986;85(1):81-7. doi: 10.1007/BF00508657.
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.
报道了一种制备直径在5至12纳米之间的胶体金颗粒的新方法。初始金颗粒群体的平均直径为5.6±0.9纳米，通过用白磷还原氯金酸制备。在胶体金颗粒存在的情况下，通过用白磷还原氯金酸使颗粒直径通过生长而增加。在大肠杆菌细胞超薄冷冻切片上标记β-半乳糖苷酶，研究了这些与蛋白A偶联的金颗粒在间接免疫标记实验中的标记效率。我们证明标记效率至少取决于颗粒直径、探针浓度和制备方法。此外还表明，用这种新方法可以制备直径分布重叠较小的金颗粒群体。因此，这些金探针适用于定性双重标记实验。讨论了免疫标记的定量方面。
