Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy.

Marko, Michael; Hsieh, Chyongere; Schalek, Richard; Frank, Joachim; Mannella, Carmen · Nat Methods · 2007

basic_science · Level V

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Abstract

Cryo-electron microscopy can provide high-resolution structural information about cells and organelles in the nearly native, frozen-hydrated state. Applicability, however, is limited by difficulties encountered in preparing suitably thin, vitreously frozen biological specimens. We demonstrate, by cryo-electron tomography of Escherichia coli cells, that a focused ion beam (FIB) can be used to thin whole frozen-hydrated cells in a convenient and essentially artifact-free way.

Medical subject headings