MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge, UK.
Department of Structural Biology, Genentech, South San Francisco, CA, USA.
FEBS Open Bio. 2024 Nov;14(11):1788-1804. doi: 10.1002/2211-5463.13873. Epub 2024 Aug 15.
Electron tomography of frozen, hydrated samples allows structure determination of macromolecular complexes that are embedded in complex environments. Provided that the target complexes may be localised in noisy, three-dimensional tomographic reconstructions, averaging images of multiple instances of these molecules can lead to structures with sufficient resolution for de novo atomic modelling. Although many research groups have contributed image processing tools for these tasks, a lack of standardisation and interoperability represents a barrier for newcomers to the field. Here, we present an image processing pipeline for electron tomography data in RELION-5, with functionality ranging from the import of unprocessed movies to the automated building of atomic models in the final maps. Our explicit definition of metadata items that describe the steps of our pipeline has been designed for interoperability with other software tools and provides a framework for further standardisation.
冷冻、水合样品的电子断层扫描允许确定嵌入复杂环境中的大分子复合物的结构。只要目标复合物可以定位于嘈杂的三维断层重建中,对这些分子的多个实例的图像进行平均处理可以得到足够分辨率的结构,以便进行从头原子建模。尽管许多研究小组已经为这些任务贡献了图像处理工具,但缺乏标准化和互操作性对于该领域的新手来说是一个障碍。在这里,我们在 RELION-5 中展示了一个用于电子断层扫描数据的图像处理管道,其功能范围从未处理的电影导入到最终映射中自动构建原子模型。我们对描述管道步骤的元数据项的明确定义旨在与其他软件工具进行互操作,并为进一步的标准化提供了一个框架。
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