Visualization of Biological Complexity Resource

描述（由申请人提供）： 沃兹沃斯中心的生物复杂性可视化资源（RVBC）采用不同分辨率水平的三维（3D）电子显微镜来研究原生环境和接近原生条件下的细胞过程，并捕捉生命物质的层次组织。(i)通过与纽约州立大学奥尔巴尼分校的合作，将利用离子束铣削等新技术进一步开发冷冻水合细胞和组织切片的低剂量冷冻断层扫描，以减少切割伪影。将努力优化数据收集，比对和重建，以实现分子特征可以识别的分辨率范围。(ii)大分子相互作用将作为动态过程进行研究，通过单颗粒形式的复合物的3D冷冻成像和时间分辨成像方法的发展，使用内部项目提供的丰富的测试平台，如兰尼碱受体和核糖体。除了喷雾冷冻和使用紫外线激活笼状化合物，我们将探索使用新的纳米纤维方法设计EM网格，在与伦斯勒理工学院的联盟协议，可以作为混合平台。(iii)杂原子数被认为是在大分子组装体的单粒子重构中实现更高分辨率的单个最大剩余障碍。监督和非监督分类的一般方法将与主要专家合作开发，使用代表该领域遇到的典型问题的幻影和实验数据集。 除了推进这三个领域的技术和研究发展，这是由研究驱动的沃兹沃斯中心和外部合作，我们继续提供独特的设施的资源，以服务项目的生物社区。

DESCRIPTION (provided by applicant): The Wadsworth Center's Resource for the Visualization of Biological Complexity (RVBC) employs three-dimensional (3D) electron microscopy at different levels of resolution to study cell processes in the native context and at close-to-native conditions, and to capture the hierarchical organization of living matter. (i) Low-dose cryo-tomography of frozen-hydrated cell and tissue sections will be further developed by utilizing novel techniques such as ion beam milling, through a collaboration with SUNY Albany, to reduce cutting artifacts. Efforts will be made to optimize data collection, alignment, and reconstruction to achieve a range of resolution where molecular signatures can be recognized. (ii) Macromolecular interactions will be studied as dynamic processes by 3D cryo-imaging of complexes in single-particle form and by development of time-resolved imaging methods, using the rich test bed provided by in-house projects such as the ryanodine receptor and the ribosome. In addition to spray-freezing and use of UV-activated caged compounds, we will explore the use of novel nanofabrication methods in designing EM grids, in a consortium agreement with the Rensselaer Polytechnic Institute, that can act as mixing platforms. (iii) Heterogeneity is recognized as the single biggest remaining obstacle to the achievement of higher resolution in single-particle reconstruction of macromolecular assemblies. General methods of supervised and unsupervised classification will be developed in collaboration with lead experts, using both phantom and experimental data sets that represent typical problems encountered in the field. In addition to advancing these three areas of technological and research development, which are driven by research at the Wadsworth Center and outside collaborations, we continue to offer the unique facilities of the Resource to the Biological Community in service projects.