Sanan D A
Research Institute for Medical Biophysics, South African Medical Research Council, Parow Valley, Cape Town.
J Microsc. 1987 Sep;147(Pt 3):341-6. doi: 10.1111/j.1365-2818.1987.tb02846.x.
A simple and convenient method is described whereby cells cultured on polystyrene plastic substrata may be critical point-dried, rotary-shadowed with platinum/carbon and finally released by means of propylene oxide for examination whole in the transmission electron microscope. The technique is particularly useful for the localization of colloidal gold probes, both intra- and extra-cellularly. The problem of premature melting of polystyrene at about 307 K after prolonged exposure to carbon dioxide is solved by limiting the residence time in the critical point-drier to 1 h. Details for cleaning and mounting the released films of cells on specimen grids are given. The technique expands the application of the 'whole-mount' approach of Hopkins and co-workers (Hopkins et al., 1981; Hopkins, 1985) to cells cultured directly on polystyrene substrata, that is, in conventional plastic culture vessels.
本文描述了一种简单便捷的方法，通过该方法，培养在聚苯乙烯塑料基质上的细胞可进行临界点干燥、用铂/碳进行旋转投影，最后通过环氧丙烷释放，以便在透射电子显微镜下进行整体观察。该技术对于细胞内和细胞外胶体金探针的定位特别有用。通过将临界点干燥器中的停留时间限制在1小时，解决了长时间暴露于二氧化碳后聚苯乙烯在约307K时过早熔化的问题。文中给出了清洁和将释放的细胞薄膜安装在标本网格上的详细步骤。该技术将霍普金斯及其同事（霍普金斯等人，1981年；霍普金斯，1985年）的“整装”方法的应用扩展到直接培养在聚苯乙烯基质上的细胞，即在传统塑料培养容器中的细胞。
