title: Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy.
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.
authors.affiliation: Resource for Visualization of Biological Complexity, Wadsworth Center, Empire State Plaza, Albany, New York 12201, USA. marko@wadsworth.org
authors.forename: Michael
authors.initials: M
authors.lastname: Marko
authors.forename: Chyongere
authors.initials: C
authors.lastname: Hsieh
authors.forename: Richard
authors.initials: R
authors.lastname: Schalek
authors.forename: Joachim
authors.initials: J
authors.lastname: Frank
authors.forename: Carmen
authors.initials: C
authors.lastname: Mannella
mesh_terms: D019412:Anatomy, Cross-Sectional / Q000379:methods*; D020285:Cryoelectron Microscopy / Q000379:methods*; D004926:Escherichia coli / Q000648:ultrastructure*; D005612:Freeze Drying / Q000379:methods; D007089:Image Enhancement / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008867:Microtomy / Q000379:methods*; D011522:Protons; D013048:Specimen Handling / Q000379:methods*
chemical_list: D011522:Protons