Peters K R, Rutter G, Gschwender H H, Haller W
J Cell Biol. 1978 Aug;78(2):309-18. doi: 10.1083/jcb.78.2.309.
For high resolution labeling of influenza virus cell surface antigens on HeLa cells, an immunospecific marker is used with silica sphere cores of 13--14 nm average diameter. These markers are formed using commercially available silica sphere sols. Two other size ranges are available, 7--8 nm and 22--25 nm. The steps for chemical derivatization are described in detail. Amino and aldehyde functions are covalently introduced onto the sphere surface. Sols of these derivatized silica spheres (DSS) are physicochemically stable and therefore usable for years. Coupling of IgG to DSS followed by permeation chromatography on controlled pore glass results in size-defined immunospecific silica sphere markers (DSS-markers). Saturation labeling of cell surface antigens on HeLa cells on cover slips is obtained with the final sphere concentration of 10(14) DSS-marker/cm3 within 20 min. With usual protective conditions, the marker stability and labeling ability are preserved for months. The visibility and the fine structure of the DSS-marker on cell surfaces are shown by using transmission electron microscopy (TEM) with stereo replicas and ultrathin sections.
为了对HeLa细胞上的流感病毒细胞表面抗原进行高分辨率标记，使用了平均直径为13 - 14纳米的二氧化硅球核的免疫特异性标记物。这些标记物是使用市售的二氧化硅球溶胶形成的。还有另外两种尺寸范围可供选择，即7 - 8纳米和22 - 25纳米。详细描述了化学衍生化的步骤。氨基和醛基功能被共价引入到球表面。这些衍生化二氧化硅球（DSS）的溶胶在物理化学上是稳定的，因此可以使用数年。将IgG与DSS偶联，然后在可控孔径玻璃上进行渗透色谱，得到尺寸确定的免疫特异性二氧化硅球标记物（DSS - 标记物）。在20分钟内，最终球浓度为10(14) DSS - 标记物/cm3时，可实现盖玻片上HeLa细胞表面抗原的饱和标记。在通常的保护条件下，标记物的稳定性和标记能力可保持数月。通过使用带有立体复制品和超薄切片的透射电子显微镜（TEM），展示了细胞表面DSS - 标记物的可见性和精细结构。
