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Subtomogram averaging from cryo-electron tomograms.
Journal article

Subtomogram averaging from cryo-electron tomograms.

  • Leigh KE Max Planck Institute for Biophysics, Frankfurt am Main, Germany; Buchmann Institute for Molecular Life Sciences, Goethe University of Frankfurt, Frankfurt am Main, Germany.
  • Navarro PP Center for Cellular Imaging and Nano-Analytics (C-CINA), Biozentrum, University of Basel, Basel, Switzerland.
  • Scaramuzza S Center for Cellular Imaging and Nano-Analytics (C-CINA), Biozentrum, University of Basel, Basel, Switzerland.
  • Chen W Max Planck Institute for Biophysics, Frankfurt am Main, Germany; Buchmann Institute for Molecular Life Sciences, Goethe University of Frankfurt, Frankfurt am Main, Germany.
  • Zhang Y Max Planck Institute for Biophysics, Frankfurt am Main, Germany; Buchmann Institute for Molecular Life Sciences, Goethe University of Frankfurt, Frankfurt am Main, Germany.
  • Castaño-Díez D Center for Cellular Imaging and Nano-Analytics (C-CINA), Biozentrum, University of Basel, Basel, Switzerland; BioEM Lab, Center for Cellular Imaging and Nano-Analytics (C-CINA), Biozentrum, University of Basel, Basel, Switzerland. Electronic address: daniel.castano@unibas.ch.
  • Kudryashev M Max Planck Institute for Biophysics, Frankfurt am Main, Germany; Buchmann Institute for Molecular Life Sciences, Goethe University of Frankfurt, Frankfurt am Main, Germany. Electronic address: misha.kudryashev@biophys.mpg.de.
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  • 2019-07-22
Published in:
  • Methods in cell biology. - 2019
Algorithms
Classification
Contrast transfer function
Cryo-electron tomography
Image processing
Software
Sub-nanometer resolution
Subtomogram averaging
Tilt series alignment
Tomogram reconstruction
Cryoelectron Microscopy
Electron Microscope Tomography
Image Processing, Computer-Assisted
Imaging, Three-Dimensional
Software
English Cryo-electron tomography (cryo-ET) allows three-dimensional (3D) visualization of frozen-hydrated biological samples, such as protein complexes and cell organelles, in near-native environments at nanometer scale. Protein complexes that are present in multiple copies in a set of tomograms can be extracted, mutually aligned, and averaged to yield a signal-enhanced 3D structure up to sub-nanometer or even near-atomic resolution. This technique, called subtomogram averaging (StA), is powered by improvements in EM hardware and image processing software. Importantly, StA provides unique biological insights into the structure and function of cellular machinery in close-to-native contexts. In this chapter, we describe the principles and key steps of StA. We briefly cover sample preparation and data collection with an emphasis on image processing procedures related to tomographic reconstruction, subtomogram alignment, averaging, and classification. We conclude by summarizing current limitations and future directions of this technique with a focus on high-resolution StA.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/232475
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