Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy |
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| Authors: | Marko Michael Hsieh Chyongere Schalek Richard Frank Joachim Mannella Carmen |
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| Affiliation: | Resource for Visualization of Biological Complexity, Wadsworth Center, Empire State Plaza, Albany, New York 12201, USA. marko@wadsworth.org |
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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. |
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