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Conical Fourier shell correlation applied to electron tomograms.

C A Diebolder1, F G A Faas2, A J Koster2

  • 1Department of Molecular Cell Biology, Leiden University Medical Center, P.O. Box 9600, 2300 RC Leiden, The Netherlands; Crystal and Structural Chemistry, Bijvoet Center for Biomolecular Research, Department of Chemistry, Faculty of Science, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.

Journal of Structural Biology
|April 7, 2015
PubMed
Summary

Dual-axis electron tomography improves image resolution isotropy by reducing anisotropic effects inherent in single-axis methods. This study introduces conical Fourier shell correlation (cFSCe/o) to assess tomogram quality and compare acquisition strategies.

Keywords:
Conical Fourier shell correlationCryo-electron microscopyDual-axis cryo-electron tomographyFourier shell correlationResolution estimation

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Area of Science:

  • Structural biology
  • Microscopy techniques
  • Image processing

Background:

  • Electron tomogram resolution is limited by anisotropic effects from data collection geometry.
  • Dual-axis tilt series acquisition can mitigate these anisotropic resolution issues.

Purpose of the Study:

  • To evaluate the benefit of dose fractionation for dual-axis cryo-electron tomography.
  • To compare the resolution isotropy of single-axis versus dual-axis electron tomograms.
  • To introduce a novel metric, conical Fourier shell correlation (cFSCe/o), for assessing tomographic reconstruction quality.

Main Methods:

  • Development and application of conical Fourier shell correlation (cFSCe/o) for resolution isotropy assessment.
  • Comparison of single-axis and dual-axis (cryo-)electron tomograms using even/odd data splits.
  • Evaluation of different alignment methods for dual-axis tomograms.

Main Results:

  • Dual-axis tomograms exhibit improved resolution isotropy in the plane orthogonal to the electron beam compared to single-axis.
  • High-resolution artifacts along the tilt axis are reduced in dual-axis acquisitions.
  • Different reconstruction programs yield varying degrees of anisotropy in dual-axis tomograms.

Conclusions:

  • Dual-axis electron tomography offers enhanced resolution isotropy, particularly when dose is fractionated.
  • cFSCe/o is a valuable tool for comparing acquisition parameters, reconstruction methods, and hardware in electron tomography.
  • Optimizing dual-axis acquisition and reconstruction is crucial for achieving isotropic resolution in cryo-electron tomography.