AUTOMATED RCT/ORT DATA COLLECTION

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 使用电子显微镜的结构可视化提供了许多见解的组成，安排和机制的大分子组装。 通过将样品保存在染色剂或玻璃冰中，可以将它们成像为离散颗粒，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，并且当面对构象变化时，它经常失败。 随机锥形倾斜（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，特别是在冰上，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.