Abstract
Freeze-drying followed by heavy-metal shadowing is an established and straightforward approach to routine structural studies of dehydrated biological specimens (Nermut and Frank 1971; Kistler et al. 1977; Gross et al. 1985). Very thin specimens such as isolated membranes or isolated macromolecules can be directly adsorbed on carbon-coated grids. After rapid freezing, the grids are transferred into suitable high-vacuum equipment for freeze-drying and shadowing. After stabilizing the heavy-metal coat with carbon (C-backing), the specimens are warmed to room temperature and transferred via atmospheric conditions into the transmission electron microscope (TEM). In other words, the entire sandwich consisting of C-supporting film, biological specimen, metal coat and C-backing film is examined in the microscope.
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