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Preparation and Observation of Thick Biological Samples by Scanning Transmission Electron Tomography
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CryoSTEM tomography in biology.

Sharon G Wolf1, Michael Elbaum2

  • 1Department of Chemical Research Support, Weizmann Institute of Science, Rehovot, Israel.

Methods in Cell Biology
|July 22, 2019
PubMed
Summary

Scanning transmission electron cryo-tomography (cryo-STEM) allows 3D imaging of thick biological cells. This technique provides chemical characterization and elemental mapping of cellular structures without staining.

Keywords:
Amorphous calcium phosphateCSTETEDSEDXElectron tomographyEnergy-dispersive X-ray spectroscopyFerritinMetalloproteinMitochondrial depositsPolyphosphate bodiesScanning probe microscopyScanning transmission electron microscopy (STEM)Z contrastcryo-EMcryo-microscopy

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Area of Science:

  • Cellular and Molecular Imaging
  • Biophysics
  • Electron Microscopy

Background:

  • Electron cryo-tomography (cryo-STEM) offers 3D reconstruction of biological specimens.
  • Contrast in cryo-STEM is determined by mass, thickness, and atomic number.
  • This technique is suitable for unstained, vitrified samples up to 1μm thick.

Purpose of the Study:

  • To provide background and practical guidance for performing cryo-STEM tomography.
  • To highlight the capabilities of cryo-STEM for chemical characterization and mass mapping of cells.
  • To explain the application of energy-dispersive X-ray spectroscopy (EDX) in conjunction with cryo-STEM.

Main Methods:

  • Utilizing scanning transmission electron microscopy (STEM) for tomographic reconstruction.
  • Applying cryo-fixation techniques to preserve cellular structure in a vitrified state.
  • Performing energy-dispersive X-ray spectroscopy (EDX) for elemental analysis.

Main Results:

  • Achieved 3D reconstructions of intact prokaryotic and eukaryotic cells.
  • Demonstrated quantifiable mass mapping and chemical characterization based on contrast.
  • Successfully identified elemental composition in sub-cellular components and mineral deposits using EDX.

Conclusions:

  • Cryo-STEM tomography is a powerful tool for high-resolution 3D imaging of thick biological specimens.
  • The technique enables detailed chemical and elemental analysis of cellular structures.
  • Cryo-STEM provides valuable insights into the composition and organization of biological systems.