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)使用不同分辨率水平的三维(3D)电子显微镜来研究自然环境和接近自然条件下的细胞过程，并捕捉生物物质的分层组织。(I)将通过与纽约州立大学奥尔巴尼分校合作，利用离子束研磨等新技术，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，以实现可识别分子特征的一系列分辨率。(2)将利用兰尼定受体和核糖体等内部项目提供的丰富的试验台，通过单粒子形式的复合体的三维冷冻成像和时间分辨成像方法的开发，将大分子相互作用作为动态过程进行研究。除了喷雾冷冻和使用紫外线激活的笼状化合物外，我们还将与伦斯勒理工学院达成联盟协议，探索使用新的纳米制造方法来设计可用作混合平台的EM网格。(3)异质性被认为是在大分子组装体的单粒子重建中实现更高分辨率的唯一最大剩余障碍。将与牵头专家合作，利用代表实地遇到的典型问题的模型和实验数据集，制定监督和非监督分类的一般方法。 除了推进这三个领域的技术和研究开发，这三个领域是由沃兹沃斯中心的研究和外部合作推动的，我们继续为生物社区的服务项目提供资源的独特设施。

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.