Freeze-substitution studies of bacteria

Typically, models of bacterial structure combine biochemical data obtained from bulk analyses of cell populations with electron microscopic observation of individual cells. Recent development of a battery of cryotechniques specific for biological electron microscopy have begun to supercede routine procedures such as conventional thin sectioning. One of these cryotechniques, freeze-substitution, combines the advantages of ultrarapid freezing with standard microtomy methods. This technique is particularly well suited to the examination of bacterial structure and has yielded additional ultrastructural information consistent with biochemical data but often challenging models of cell structure obtained from conventional microscopical methods. In addition to retaining more accurately the spatial distribution of cell components, freeze-substitution has been successfully combined with immunochemical labelling techniques and has enabled identification and localization of specific molecules both within the cell and on the cell surface. In this review, I describe current ideas on bacterial ultrastructure, modified in accordance with new data obtained from recent freeze-substitution studies.