ABSTRACT
Efforts to study unstained, fully hydrated thin film biological specimens in vitreous ice by TEM started some time ago (Fernandez-Moran, 1960; Hutchinson et al., 1978; Taylor and Glaeser, 1974, 1976). However, such approaches were not realistically achieved until the development of a method for rapidly freezing thin aqueous films in the early 1980s, by Jacques Dubochet and his colleagues (Dubochet and McDowall, 1981; Dubochet et al., 1982). Biological materials could then routinely be prepared as frozen-hydrated specimens, with untreated material (unfixed and unstained) embedded within a thin layer of vitreous/non-crystalline ice.
