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Improved applicability and robustness of fast cryo-electron tomography data acquisition.
Journal article

Improved applicability and robustness of fast cryo-electron tomography data acquisition.

  • Eisenstein F Department of Biology, Institute of Molecular Biology & Biophysics, Eidgenössische Technische Hochschule Zürich, Otto-Stern-Weg 5, 8093 Zürich, Switzerland.
  • Danev R Graduate School of Medicine, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
  • Pilhofer M Department of Biology, Institute of Molecular Biology & Biophysics, Eidgenössische Technische Hochschule Zürich, Otto-Stern-Weg 5, 8093 Zürich, Switzerland. Electronic address: pilhofer@biol.ethz.ch.
  • 2019-08-20
Published in:
  • Journal of structural biology. - 2019
Calibration
Cryo-EM
Cryo-electron tomography
Data acquisition
Electron cryotomography
K2
K3
Throughput
Tilt series
Algorithms
Cryoelectron Microscopy
Electron Microscope Tomography
Escherichia coli
Ribosomes
English The power of cryo-electron tomography (cryoET) lies in its capability to characterize macromolecules in their cellular context. Structure determination by cryoET, however, is time-consuming compared to single particle approaches. A recent study reported significant acceleration of data acquisition by a fast-incremental single-exposure (FISE) tilt series scheme. Here we improved the method and evaluated its efficiency and performance. We show that (1) FISE combined with the latest generation of direct electron detectors speeds up collection considerably, (2) previous generation (pre-2017) double-tilt axis Titan Krios holders are also suitable for FISE data acquisition, (3) x, y and z-specimen shifts can be compensated for, and (4) FISE tilt series data can generate averages of sub-nanometer resolution. These advances will allow for a widespread adoption of cryoET for high-throughput in situ studies and high-resolution structure determination across different biological research disciplines.
Language
  • English
Open access status
hybrid
Identifiers
Persistent URL
https://sonar.ch/global/documents/185475
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