Improved applicability and robustness of fast cryo-electron tomography data acquisition
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Author / Producer
Date
2019-11-01
Publication Type
Journal Article
ETH Bibliography
yes
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Abstract
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.
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Publication status
published
External links
Editor
Book title
Journal / series
Volume
208 (2)
Pages / Article No.
107 - 114
Publisher
Elsevier
Event
Edition / version
Methods
Software
Geographic location
Date collected
Date created
Subject
Cryo-electron tomography; Electron cryotomography; Cryo-EM; Tilt series; Data acquisition; Throughput; K2; K3; Calibration
Organisational unit
09463 - Pilhofer, Martin / Pilhofer, Martin
Notes
Funding
679209 - Multiscale model of bacterial cell-cell interactions (EC)
Related publications and datasets
Is referenced by: http://hdl.handle.net/20.500.11850/639504