title: Cryo-scanning transmission electron tomography of vitrified cells.
abstract: Cryo-electron tomography (CET) of fully hydrated, vitrified biological specimens has emerged as a vital tool for biological research. For cellular studies, the conventional imaging modality of transmission electron microscopy places stringent constraints on sample thickness because of its dependence on phase coherence for contrast generation. Here we demonstrate the feasibility of using scanning transmission electron microscopy for cryo-tomography of unstained vitrified specimens (CSTET). We compare CSTET and CET for the imaging of whole bacteria and human tissue culture cells, finding favorable contrast and detail in the CSTET reconstructions. Particularly at high sample tilts, the CSTET signals contain more informative data than energy-filtered CET phase contrast images, resulting in improved depth resolution. Careful control over dose delivery permits relatively high cumulative exposures before the onset of observable beam damage. The increase in acceptable specimen thickness broadens the applicability of electron cryo-tomography.
authors: Department of Chemical Research Support, Weizmann Institute of Science, Rehovot, Israel.
authors: Sharon Grayer
authors: SG
authors: Wolf
authors: 1] Ernst Ruska-Centre for Microscopy and Spectroscopy with Electrons, Forschungszentrum Jülich GmbH, Jülich, Germany. [2] Peter Grünberg Institute, Forschungszentrum Jülich GmbH, Jülich, Germany.
authors: Lothar
authors: L
authors: Houben
authors: Department of Materials and Interfaces, Weizmann Institute of Science, Rehovot, Israel.
authors: Michael
authors: M
authors: Elbaum
mesh_terms: D016960:Agrobacterium tumefaciens / Q000166:cytology*; D000818:Animals; D002244:Carbon / Q000737:chemistry*; D007053:Ice / Q000032:analysis*; D017348:Microscopy, Electron, Scanning Transmission / Q000379:methods*; D058989:Vitrification
chemical_list: D007053:Ice; D002244:Carbon