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Electron cryo‐microscopy of vitrified bulk biological specimens: ideal and real structures of water—lipid phases

Lipid‐water mixtures were studied by X‐ray cryodiffraction in order to assess the structural changes during freezing. We show that the water of aqueous lipid phases, in the concentration range of 10–30% (water weight/total weight), is vitrified by high‐pressure freezing. Vitrified lipid phases can be cryo‐sectioned and imaged by electron cryomicroscopy. Both the ideal or average and the real or local structures of the lipid mixtures can be studied at a resolution better than 2 nm. While the average structure of the lipid phases is in good agreement with that determined by X‐ray diffraction, the local structure reveals features that might play an important role in the function of biological membranes such as in endo‐ and exocytosis.