Tools for High-Throughput High-Resolution Three Dimensional Electron Microscopy

高通量高分辨率三维电子显微镜工具

• 批准号: 8817532
• 负责人: SCOTT M STAGG
• 金额: $ 28.78万
• 依托单位: FLORIDA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-30 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8817532
• 关键词: Automatic Data Processing; Biological; Classification; Complex; Computer software; Crystallography; Data; Data Collection; Disease; Electron Microscopy; Feedback; Filament; Generations; Health; Human; Image; Microscope; Molecular; Preparation; Process; Proteins; Research; Research Infrastructure; Research Personnel; Resolution; Sampling; Series; Side; Software Tools; Specimen; Structure; Techniques; Time; Virus; base; computerized data processing; cryogenics; data acquisition; density; electron tomography; human disease; improved; molecular assembly/self assembly; particle; protein complex; protein structure; public health relevance; reconstruction; screening; software development; tomography; tool

DESCRIPTION (provided by applicant): Three dimensional electron microscopy (3DEM) is a set of techniques for determining the structures of large proteins and protein complexes. It has been shown that cryogenic EM (cryo-EM) has the ability to determine structures to high enough resolution to unambiguously place side chains in the EM density. Though cryo-EM has the potential for atomic resolution, a number of hurdles are present in order to achieve high-throughput structure determination of low-symmetry and no symmetry complexes. Here we propose to develop software tools for high-throughput high-resolution 3DEM. In the first aim, we propose to develop software tools for single particle 3DEM including software for automated specimen screening, automated data acquisition parameter optimization, and an infrastructure for processing helical samples. In the second aim, we propose to develop tools for tomographic data collection and processing including acquisition of dual axis tilt series, and developing and infrastructure for processing tomographic subvolumes. This research will benefit human health by facilitating the determination of protein structures that are relevant for human disease.

描述（由申请人提供）：三维电子显微镜（3DEM）是一组用于确定大蛋白质和蛋白质复合物结构的技术。已经表明，低温EM（Cryo-EM）具有确定足够高分辨率的结构，以明确地将侧链放置在EM密度中。尽管冷冻EM具有原子分辨率的潜力，但仍存在许多障碍，以实现低对称性的高通量结构的测定和无对称复合物。在这里，我们建议开发用于高通量高分辨率3DEM的软件工具。在第一个目标中，我们建议为单个粒子3DEM开发软件工具，包括用于自动样品筛选的软件，自动数据采集参数优化以及用于处理螺旋样品的基础架构。在第二个目的中，我们建议开发用于层析成像数据收集和处理的工具，包括获得双轴倾斜系列，以及用于处理层析成像亚参数的开发和基础架构。这项研究将通过促进与人类疾病相关的蛋白质结构的确定来使人类健康受益。