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Benchmarking tomographic acquisition schemes for high-resolution structural biology
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Benchmarking tomographic acquisition schemes for high-resolution structural biology
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Journal Article
Benchmarking tomographic acquisition schemes for high-resolution structural biology
2020
Overview
Cryo electron tomography with subsequent subtomogram averaging is a powerful technique to structurally analyze macromolecular complexes in their native context. Although close to atomic resolution in principle can be obtained, it is not clear how individual experimental parameters contribute to the attainable resolution. Here, we have used immature HIV-1 lattice as a benchmarking sample to optimize the attainable resolution for subtomogram averaging. We systematically tested various experimental parameters such as the order of projections, different angular increments and the use of the Volta phase plate. We find that although any of the prominently used acquisition schemes is sufficient to obtain subnanometer resolution, dose-symmetric acquisition provides considerably better outcome. We discuss our findings in order to provide guidance for data acquisition. Our data is publicly available and might be used to further develop processing routines.
Here the authors systematically benchmark cryo-electron tomography acquisition schemes to optimize the attainable resolution for subtomogram averaging, and find that dose-symmetric acquisition with even angular sampling provides a better outcome than most currently used acquisition schemes.
Publisher
Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
Subject
/ 631/535
/ Cryoelectron Microscopy - methods
/ Cryoelectron Microscopy - standards
/ Electron Microscope Tomography - methods
/ Electron Microscope Tomography - standards
/ HIV
/ Human immunodeficiency virus
/ Humanities and Social Sciences
/ Macromolecular Substances - chemistry
/ Macromolecular Substances - ultrastructure
/ Molecular Biology - standards
/ Science
