TRD4: Design and Engineering of MicroED hardware for nanocrystallization and time resolved studies

TRD4：用于纳米结晶和时间分辨研究的 MicroED 硬件的设计和工程

• 批准号: 10641827
• 负责人: JOHN Charles SPENCE
• 金额: $ 21.05万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10641827
• 关键词: Acceleration; Affect; Alcohol consumption; Alcohols; Amino Acids; Area; Binding; Biological; Biological Assay; Carbon; Charge; Chemicals; Chemistry; Complex; Crystallization; Data; Data Collection; Detergents; Development; Diffusion; Electron Diffraction Microscopy; Engineering; Environment; Equilibrium; Equipment; Freezing; Growth; Humidity; Hydrophobicity; Lead; Ligands; Methodology; Methods; Microscope; Modality; Molecular Structure; Oils; Pharmaceutical Preparations; Play; Preparation; Procedures; Proteins; Protocols documentation; Reaction; Robot; Roentgen Rays; Role; Sampling; Solvents; Structure; Surface; System; Techniques; Thinness; Time; Work; X-Ray Crystallography; X-Ray Medical Imaging; aqueous; beamline; chemical property; design; electron diffraction; experimental study; imaging facilities; improved; macromolecular assembly; nano; nanolitre; prototype; receptor; response; screening; small molecule; success; tool

TRD4. Design and Engineering of MicroED hardware for nanocrystallization and time resolved studies – Spence (Lead) Summary The initial development of microcrystal electron diffraction (MicroED), and subsequent improvements to the technique, have been driven by new data collection methods and hardware for controlling the microscope. Of particular importance is turning MicroED into a high-throughput method with time-resolved capability to be able to rapidly determine time-dependent structure-function relation of functioning macomolecular complexes. This will propel the technique forward, but it requires additional design and engineering efforts in order to make these desired features a reality. Here, we will focus on three MicroED engineering aims: (1) the development of high throughput on-grid crystallization equipment to increase the efficiency and success rate of MicroED experiments, (2) the adaptation and design of new equipment and methods to conduct time-resolved MicroED experiments, (3) the design of a multi-functional grid surface, or “lab on a grid” for the streamlined preparation of small molecule samples for MicroED structure determination. The aims of this project are 1. Development of high throughput on-grid crystallization for MicroED; 2. New sample preparation methodology for time-resolved MicroED; 3. “Lab on a grid” for MicroED analysis of small molecules. Overall, we will design, engineer, and implement new tools and techniques for three major aspects of MicroED. These new tools will greatly accelerate the throughput capabilities of the MicroED and improve the quality of collected data. Additionally, new experimental modalities will be made possible through the work in Aim 2, opening the door to precise time-resolved studies by MicroED.

TRD4.用于纳米晶化和时间分辨研究的MicroED硬件的设计和工程 斯宾塞(领衔) 摘要 微晶电子衍射(MicroED)的初步发展，以及后来对微晶电子衍射的改进 新的数据收集方法和控制显微镜的硬件推动了这一技术的发展。的 特别重要的是将MicroED转变为具有时间分辨能力的高通量方法 快速测定功能性大分子络合物随时间变化的结构-功能关系。这 将推动技术向前发展，但它需要额外的设计和工程努力才能使这些 想要的特征变成了现实。在这里，我们将集中讨论三个微电火花工程目标：(1)发展高 通过在线晶化设备提高MicroED实验的效率和成功率， (2)适应和设计新的设备和方法以进行时间分辨的MicroED实验， (3)用于流线型小分子制备的多功能网格面的设计，或“网格上的实验室” 用于MicroED结构测定的样品。该项目的目标是：1.发展高吞吐量 时间分辨微电子能谱样品制备新方法；“实验室 用于小分子的微电子能谱分析。总体而言，我们将设计、设计和实施新工具 以及MicroED的三个主要方面的技术。这些新工具将极大地提高吞吐量 提高微型ED的能力，提高所收集数据的质量。此外，新的实验模式 将通过AIM 2的工作成为可能，为MicroED进行精确的时间分辨研究打开了大门。