Focused-ion-beam thinning of frozen-hydrated biological specimens for cryo-electron microscopy.
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.
Marko
Michael
M
Resource for Visualization of Biological Complexity, Wadsworth Center, Empire State Plaza, Albany, New York 12201, USA. marko@wadsworth.org
Hsieh
Chyongere
C
Schalek
Richard
R
Frank
Joachim
J
Mannella
Carmen
C
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*
D011522:Protons