Benchmarking tomographic acquisition schemes for high-resolution structural biology.,Nature Communications

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Benchmarking tomographic acquisition schemes for high-resolution structural biology.
Nature Communications ( IF 14.7 ) Pub Date : 2020-02-13 , DOI: 10.1038/s41467-020-14535-2
Beata Turoňová ₁ , Wim J H Hagen ₁ , Martin Obr _{2,

3} , Shyamal Mosalaganti ₁ , J Wouter Beugelink _{1,

4,

5} , Christian E Zimmerli _{1,

6} , Hans-Georg Kräusslich ₂ , Martin Beck _{1,

7}

Affiliation

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.

中文翻译：

高分辨率结构生物学的断层摄影获取方案的基准。

低温电子层析成像和随后的子图平均化技术是一种强大的技术，可以在自然背景下对大分子复合物进行结构分析。尽管原则上可以获得接近原子的分辨率，但尚不清楚单个实验参数如何有助于达到分辨率。在这里，我们使用未成熟的HIV-1晶格作为基准样本，以优化可实现的子图平均化分辨率。我们系统地测试了各种实验参数，例如投影顺序，不同的角度增量和Volta相板的使用。我们发现，尽管任何最常用的采集方案都足以获得亚纳米分辨率，但剂量对称采集可提供更好的结果。我们讨论我们的发现，以便为数据获取提供指导。我们的数据可公开获得，并可用于进一步开发处理程序。

更新日期：2020-02-13

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