Takizawa T, Robinson J M
Department of Cell Biology, Neurobiology and Anatomy, Ohio State University, Columbus.
J Histochem Cytochem. 1994 Dec;42(12):1615-23. doi: 10.1177/42.12.7983362.
We present a new application for the use of small immunogold particles (approximately 1.4-nm diameter) for ultrastructural immunocytochemistry. These small gold particles have been used on ultra-thin cryosections in conjunction with a silver enhancement procedure that does not degrade ultrastructural detail. We have used the human neutrophil as a model system, in which known protein markers of two different cytoplasmic granules were localized, in the development of this procedure. The 1.4-nm immunogold particles coupled with silver enhancement yield intense labeling for localization of lactoferrin, a marker for the specific granules, and myeloperoxidase, a marker for the azurophil granules. Double labeling in which one antigen was visualized with 1.4-nm gold and silver enhancement and a second antigen was detected with colloidal gold-IgG on the same ultra-thin cryosection was successfully achieved. We also show that 1.4-nm diameter immunogold particles penetrate into cryosectioned neutrophils to a greater extent than 5-nm or 10-nm immunogold probes. These results show that small immunogold particles, along with silver enhancement, are a useful addition to the immunolabeling methods available for use with ultra-thin cryosections.
我们展示了一种将小免疫金颗粒（直径约1.4纳米）用于超微结构免疫细胞化学的新应用。这些小金颗粒已用于超薄冷冻切片，并结合一种不会破坏超微结构细节的银增强程序。在开发此程序的过程中，我们以人类中性粒细胞作为模型系统，定位了两种不同细胞质颗粒的已知蛋白质标记物。与银增强相结合的1.4纳米免疫金颗粒能产生强烈的标记信号，用于定位乳铁蛋白（一种特异性颗粒的标记物）和髓过氧化物酶（一种嗜天青颗粒的标记物）。在同一超薄冷冻切片上成功实现了双重标记，其中一种抗原用1.4纳米金和银增强进行可视化，另一种抗原用胶体金 - IgG进行检测。我们还表明，直径1.4纳米的免疫金颗粒比5纳米或10纳米的免疫金探针更能深入冷冻切片的中性粒细胞。这些结果表明，小免疫金颗粒与银增强一起，是可用于超薄冷冻切片的免疫标记方法中的一项有用补充。
