Visualization of Biological Complexity Resource

沃兹沃斯中心的生物复杂性可视化资源（RVBC）采用三维 (3D)电子显微镜在不同的分辨率水平，以研究细胞过程中的自然 背景和接近原生条件，并捕捉生命物质的层次结构，（i） 冷冻水化细胞和组织切片的低剂量冷冻断层扫描将通过利用 通过与纽约州立大学奥尔巴尼分校的合作，采用离子束铣削等新技术来减少切割 藏物将努力优化数据收集、调整和重建，以实现一系列 （ii）将研究大分子相互作用， 通过单粒子形式的复合物的3D冷冻成像和通过时间分辨的 成像方法，使用内部项目提供的丰富测试平台，如ryanodine 受体和核糖体。除了喷雾冷冻和使用紫外线激活笼化合物，我们将 探索在设计EM网格中使用新的纳米纤维方法， 伦斯勒理工学院，可以作为混合平台，（三）Heteroadministration被认为是 在单粒子重建中实现更高分辨率的一个最大的障碍， 大分子组装体监督和非监督分类的一般方法将 与主要专家合作开发，使用模型和实验数据集， 现场遇到的典型问题。 除了推进这三个技术和研究发展领域外， 研究在沃兹沃斯中心和外部合作，我们继续提供独特的设施， 生物社区服务项目的资源。

The Wadsworth Center's Resource for the Visualization of Biological Complexity (RVBC) employs threedimensional (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 timeresolved 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.