AUTOMATED RCT/ORT DATA COLLECTION

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 利用电子显微镜的结构可视化提供了许多关于大分子组装的组成、排列和机理的见解。通过将样品保存在污渍或玻璃冰中，可以将它们成像为离散的颗粒，并从这些图像生成三维结构。这种单粒子方法有两个主要缺点：它需要一个初始模型来将2D数据重建成3D体积；当面临构象变化时，它经常失败。随机圆锥倾斜(RCT)和正交倾斜(OTR)是为克服这些问题而开发的方法，但所需的数据收集，特别是在冰层中，是困难和繁琐的。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Structural visualization using electron microscopy has provided many insights into the composition, arrangement and mechanism of macromolecular assemblies. By preserving samples in stain or vitreous ice it is possible to image them as discrete particles, and from these images generate three-dimensional structures. This 'single-particle' approach suffers from two major shortcomings; it requires an initial model to reconstitute 2D data into a 3D volume, and it often fails when faced with conformational variability. Random conical tilt (RCT) and orthogonal tilt (OTR) are methods developed to overcome these problems, but the data collection required, particularly in ice, is difficult and tedious.