300 keV FEG Electron Cryo Microscope for Single Particle Analyses

用于单粒子分析的 300 keV FEG 冷冻电子显微镜

• 批准号: 342246990
• 金额: --
• 依托单位: Universität Hamburg
• 依托单位国家: 德国
• 项目类别: Major Research Instrumentation
• 财政年份: 2017
• 资助国家: 德国
• 起止时间: 2016-12-31 至 无数据
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/342246990?language=en
• 关键词: 300; keV; FEG; Electron; Cryo

Single particle electron cryo microscopy (cryoEM) enables structural analyses of the macromolecular building blocks of cells. Probing structure-function relationships is essential for understanding basic molecular mechanisms in biology. We here aim to analyse in detail macromolecules and their dynamic changes in the course of infections – a major research focus in Hamburg. The technical advancements and break troughs of the recent years have moved the technology to a state allowing to achieve routinely three-dimensional structures in a resolution range between 2.5 und 6Å for stable protein complexes. This ‚revolution’ is based on the following factors: (1) development of robust 300keV high-resolution electron cryo microscopes with parallel illumination and robotic sample handling, (2) introduction of direct electron detectors, (3) automated data acquisition software, (4) more robust and versatile analysis software packages and (5) better computing hardware. Together these developments have moved single particle cryoEM to an efficient and crystallisation independent alternative for high-resolution structural analyses of biological macromolecules. In particular for larger more flexible macromolecular complexes with various functional conformations single particle cryoEM in combination with robust classification algorithms provides a highly attractive approach in structural biology. Demand for respective cryoEM access has risen immensely and includes now communities beyond ‘classical’ structural biologists, like biochemist and cell biologists (cf. the broad user base of the instrument).

单粒子电子冷冻显微镜（cryoEM）能够对细胞的大分子结构进行结构分析。探索结构-功能关系对于理解生物学中的基本分子机制至关重要。我们在这里的目标是详细分析大分子及其在感染过程中的动态变化--这是汉堡的一个主要研究重点。近年来的技术进步和突破已经使该技术发展到允许在2.5和6 μ m之间的分辨率范围内实现稳定蛋白质复合物的常规三维结构的状态。这场“革命”基于以下因素：（1）开发具有平行照明和机器人样品处理功能的强大的300 keV高分辨率电子冷冻显微镜，（2）引入直接电子检测器，（3）自动数据采集软件，（4）更强大和多功能的分析软件包和（5）更好的计算硬件。这些发展共同推动了单粒子cryoEM成为生物大分子高分辨率结构分析的有效和独立于结晶的替代方案。特别是对于具有各种功能构象的更大更灵活的大分子复合物，单粒子cryoEM与鲁棒的分类算法相结合，在结构生物学中提供了一种非常有吸引力的方法。对相应cryoEM访问的需求已经大大增加，现在包括“经典”结构生物学家以外的社区，如生物化学家和细胞生物学家（参见。仪器的广泛用户基础）。