Development and Commercialization of a Sample Preparation System for Time Resolved Cryo-Electron Microscopy

时间分辨冷冻电子显微镜样品制备系统的开发和商业化

• 批准号: 10081915
• 负责人: JOACHIM FRANK
• 金额: $ 25.21万
• 依托单位: HUMMINGBIRD PRECISION MACHINE COMPANY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10081915
• 关键词: Biological; Biological Process; Cone; Consumption; Cost efficiency; Cryoelectron Microscopy; Data; Development; Dimensions; Documentation; Electron Beam; Electrons; Environmental Monitoring; Ethane; Evaluation; Evolution; Freezing; Gases; Government; Humidity; Hydration status; Image; Imaging Techniques; Kinetics; Liquid substance; Measures; Methods; Microfluidics; Molecular Structure; Names; Pathway interactions; Persons; Phase; Population; Population Distributions; Positioning Attribute; Preparation; Principal Investigator; Process; Production; Reaction; Reaction Time; Recipe; Reproducibility; Research; Research Personnel; Resolution; Running; Salmon; Sampling; Small Business Technology Transfer Research; Speed; Standardization; Structure; System; Techniques; Temperature; Testing; Time; Universities; Work; X-Ray Crystallography; biological research; biological systems; commercialization; cost; design; experimental study; improved; millisecond; operation; particle; preservation; pressure; prototype; software development; temporal measurement; touchscreen; user-friendly

Project Summary Project Title: Development and Commercialization of a Sample Preparation System for Time Resolved Cryo-Electron Microscopy Company Name: Hummingbird Precision Machine Co., dba Hummingbird Scientific Principal Investigator: Norman Salmon Summary: Cryo-electron microscopy (cryo-EM) is a powerful technique for imaging biological molecules in which samples are preserved in a frozen, hydrated state before imaging using an electron beam. The technique provides an alternative to X-ray crystallography for structural characterization of biological samples that are not crystallizable or do not crystallize in their native state. While cryo- EM sample preparation technique has become more efficient in recent years with the development of automatic sample vitrification systems to perform grid preparation, the field of cryo-EM is hindered by the inability to perform time-resolved experiments on a sufficiently fast time scale to observe the progression of many biological reactions due to the inherent lag of traditional blotting methods. The Frank group at Columbia University has developed a lab- prototype of a system to enable this missing capability and enables millisecond time control over the reaction time of sampling biological reactions in the grid preparation process. The purpose of this Fast-Track STTR proposal is to develop the time-resolved system built by the Frank group into a commercial production unit and launch it as a product. Initial Phase I milestones in the proposed work will focus on building a prototype with added control of reaction parameters to determine necessary control variables in the system and to optimize sample grid preparation and time resolution. Further work in Phase II of this Fast-Track proposal will create a streamlined, user-friendly production unit which will maintain the necessary controls as determined in initial milestones, while optimizing manufacturing time and reducing overall production costs. The resulting system will add another dimension to the field of cryo-EM, providing users with the capability to capture transient intermediates in reaction progression studies and further clarifying proposed reaction mechanisms with time resolution.

项目摘要 项目名称：时间准备系统的开发和商业化 分辨的冷冻电子显微镜 公司名称：DBA Hummingbird Scientific Sc​​ientific 首席研究员：诺曼鲑鱼 概括： 冷冻电子显微镜（Cryo-EM）是成像生物分子的强大技术 在使用电子束进行成像之前，将哪些样品保存在冷冻的水合状态下。 该技术为X射线晶体学提供了替代方案，用于结构表征 无法结晶或不结晶的生物样品。而冷冻 - 近年来，EM样品制备技术已经变得更加有效 开发自动样品玻璃化系统以执行网格准备， 由于无法在足够快的快速上执行时间分辨的实验，因此阻碍了Cryo-EM 时间尺度观察由于许多生物反应的进展 传统印迹方法。哥伦比亚大学的弗兰克集团已经开发了一个实验室 系统的原型，以实现这种缺失的功能并实现毫秒的时间控制 在网格制备过程中抽样生物反应的反应时间。 这项快速通道的STTR提案的目的是开发由该系统建立的时间分辨系统 弗兰克集团进入商业生产部门并将其作为产品启动。初始阶段I。 拟议工作中的里程碑将集中于建立一个原型，并增加了反应的控制 参数以确定系统中必要的控制变量并优化样品网格 准备和时间分辨率。在此快速提案的第二阶段中进一步工作将创建一个 流线型，用户友好的生产单元将维持必要的控件 在初始里程碑中确定，同时优化制造时间并减少整体 生产成本。最终的系统将为Cryo-Em的字段添加另一个维度， 为用户提供捕获反应进程中瞬态中间体的能力 研究并进一步澄清提出的反应机制，并通过时间分辨率进行研究。