Platinum-shadowed, whole-mounts for transmission electron microscopy of cells cultured on plastic.
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.
Research Institute for Medical Biophysics, South African Medical Research Council, Parow Valley, Cape Town.
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Sanan
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D046508:Culture Techniques / Q000379:methods*; D004727:Endothelium / Q000166:cytology / Q000648:ultrastructure; D005347:Fibroblasts / Q000648:ultrastructure; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods*; D010984:Platinum; D011137:Polystyrenes
D011137:Polystyrenes; D010984:Platinum