Automated batch fiducial-less tilt-series alignment in Appion using Protomo.
Automated batch fiducial-less tilt-series alignment in Appion using Protomo.
复制标题
使用Protomo在APPION中自动批量无基于基准的倾斜系列。
DOI：
10.1016/j.jsb.2015.10.003
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发表时间：
2015-11
影响因子：
3
通讯作者：
Stagg SM
中科院分区：
文献类型：
--
作者：
Noble AJ;Stagg SM
电子断层扫描领域从基于标记的倾斜序列比对的手动和半自动方法中获益良多，这些方法允许对大量原位细胞结构以及单个蛋白质复合物的大分子结构进行结构测定。能够检测单个电子的互补金属氧化物半导体探测器的出现，使低剂量、高对比度图像的收集成为可能，为可靠的基于相关的倾斜串联校准打开了大门。在这里，我们提出了一套自动化的，基于相关性的倾斜序列校准，对比度传递函数（CTF）校正和重建工作流程，用于与Appion/Leginon包结合使用，主要针对低温电子显微镜的自动化结构确定。
The field of electron tomography has benefited greatly from manual and semi-automated approaches to marker-based tilt-series alignment that have allowed for the structural determination of multitudes of in situ cellular structures as well as macromolecular structures of individual protein complexes. The emergence of complementary metal-oxide semiconductor detectors capable of detecting individual electrons has enabled the collection of low dose, high contrast images, opening the door for reliable correlation-based tilt-series alignment. Here we present a set of automated, correlation-based tilt-series alignment, contrast transfer function (CTF) correction, and reconstruction workflows for use in conjunction with the Appion/Leginon package that are primarily targeted at automating structure determination with cryogenic electron microscopy.