Cryo-scanning transmission electron tomography of vitrified cells

@article{Wolf2014CryoscanningTE,
  title={Cryo-scanning transmission electron tomography of vitrified cells},
  author={Sharon Grayer Wolf and Lothar Houben and Michael Elbaum},
  journal={Nature Methods},
  year={2014},
  volume={11},
  pages={423-428}
}
Cryo-electron tomography (CET) of fully hydrated, vitrified biological specimens has emerged as a vital tool for biological research. [...] Key Result 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.Expand

Paper Mentions

Elemental Analysis and Cryo-STEM Tomography of Vitrified Cells
Cryo-electron tomography (CET) has provided in recent years unparalleled sub-nm views into cellular ultrastructure for unfixed, unstained specimens in as-close-to-native state as possible. ImageExpand
Cryo-STEM Tomography of Intact Vitrified Fibroblasts
Cryo-tomography of intact, vitrified cells provides a three dimensional view of their structure and organization in a snapshot of the living state. Lacking heavy metal stains, tilt series images areExpand
Quantitative Cryo-Scanning Transmission Electron Microscopy of Biological Materials.
  • M. Elbaum
  • Materials Science, Medicine
  • Advanced materials
  • 2018
TLDR
The extension of STEM to cryomicroscopy and tomography of cells and macromolecules is summarized herein. Expand
Dose-Efficient Cryo-STEM Imaging of Vitrified Biological Samples
Cryo-transmission electron microscopy (cryo-TEM) has enabled new insights into biology as it allows the study of specimens preserved in their near-native state through vitrification. Cryo-scanningExpand
Cryo-scanning transmission electron tomography of biological cells
The electron microscope has made paramount contributions to understanding the structure of biological molecules, cells, and tissues. In general, the most faithful preservation of biological specimensExpand
CryoSTEM tomography in biology.
TLDR
This chapter provides basic background and practical information for performing cryo-STEM tomography on vitrified biological cells. Expand
Three-dimensional deconvolution processing for STEM cryotomography
TLDR
It is shown here that deconvolution leads to similar improvements in volume data reconstructed from cryoscanning transmission electron tomography (CSTET), namely a dramatic in-plane noise reduction and improved representation of features in the axial dimension. Expand
Exploring the theoretical basis and limitations of cryo-STEM tomography for thick biological specimens.
TLDR
This work forms an imaging model based on a Boltzmann transport equation, complemented by Monte Carlo simulations, and proposes to generalize the on-axis bright field detector to collect at angles well beyond the illumination cone to justify the generation of 3D images for micron thicknesses and beyond. Expand
Expanding horizons of cryo-tomography to larger volumes.
  • M. Elbaum
  • Biology, Medicine
  • Current opinion in microbiology
  • 2018
TLDR
New imaging modalities are now available to extendcryo-tomography to thicker specimens: cryo-scanning transmission electron tomography (CSTET), soft X-raytomography (SXT), and serial surface imaging using the focused ion beam- scanning electron microscope (FIB-SEM). Expand
Preparing samples from whole cells using focused-ion-beam milling for cryo-electron tomography
TLDR
This protocol describes how to prepare thin specimens using FIB milling from frozen cells on grids, which enables direct structural analysis of biomolecules in their native environments, i.e., cells. Expand
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