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首页> 外文期刊>Acta crystallographica. Section D, Structural biology >High-resolution structure determination using high-throughput electron cryo-tomography
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High-resolution structure determination using high-throughput electron cryo-tomography

机译：高分辨率结构确定使用高通量电子cryo-tomography

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摘要

Tomographic reconstruction of frozen-hydrated specimens followed by extraction and averaging of sub-tomograms has successfully been used to determine the structure of macromolecules in their native environment at resolutions that are high enough to reveal molecular level interactions. The low throughput characteristic of tomographic data acquisition combined with the complex data-analysis pipeline that is required to obtain high-resolution maps, however, has limited the applicability of this technique to favorable samples or to resolutions that are too low to provide useful mechanistic information. Recently, beam image-shift electron cryo-tomography (BISECT), a strategy to significantly accelerate the acquisition of tilt series without sacrificing image quality, was introduced. The ability to produce thousands of high-quality tilt series during a single microscope session, however, introduces significant bottlenecks in the downstream data analysis, which has so far relied on specialized pipelines. Here, recent advances in accurate estimation of the contrast transfer function and self-tuning exposure-weighting routines that contribute to improving the resolution and streamlining the structure-determination process using sub-volume averaging are reviewed. Ultimately, the combination of automated data-driven techniques for image analysis together with high-throughput strategies for tilt-series acquisition will pave the way for tomography to become the technique of choice for in situ structure determination.
机译：层析重建frozen-hydrated标本的提取和平均紧随其后sub-tomograms已成功被用来确定结构的大分子在决议,其原生环境高到足以揭示分子水平交互。层析数据采集的结合复杂的数据分析管道是必需的然而,获取高分辨率的地图限制了该技术的适用性良好的样品或决议,低提供有用的信息。最近,电子光束图像偏移cryo-tomography(平分),一个策略显著加快收购倾斜系列不牺牲图像质量,介绍了。在一个高质量的倾斜系列然而,显微镜,介绍在下游数据的重要瓶颈分析,到目前为止依靠专业管道。对比传递函数和估计自调优exposure-weighting例程,为提高分辨率和做出贡献简化结构测定过程使用sub-volume平均进行了综述。最终,自动化的结合数据驱动的图像分析技术结合高通量的策略tilt-series收购铺平道路断层扫描成为首选的技术原位结构的决心。

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