I-Corps: A freezable fluid cell for high-resolution cryo-electron microscopy

I-Corps：用于高分辨率冷冻电子显微镜的可冷冻流体池

• 批准号: 1844868
• 负责人: Rebecca Menapace
• 金额: $ 5万
• 依托单位: Brandeis University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2020-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1844868&HistoricalAwards=false
• 关键词: Corps; freezable; fluid; cell; resolution

The broader impact and commercial potential of this I-Corps project are as follows: Cryo-Electron Microscopy (Cryo-EM) is a method for obtaining 3D structures of biological molecules. The method has growing importance in basic life science research and in the pharmaceutical sector for defining how drugs interact with their targets. Cryo-EM as a whole has matured into a powerful tool in recent years, culminating in the 2017 Nobel Prize in Chemistry, but the process of protein 'sample preparation' for 3D study has not advanced appreciably and is widely considered to represent the major bottleneck in the overall method. This bottleneck also presents a major commercial opportunity. The present project advances new nanofluidic-based technology aimed at addressing this bottleneck. Successful development of this technology will yield a consumable product ready for commercialization. Fulfillment of this goal will greatly advance and open new possibilities in cryo-EM structure determination. The resulting acceleration and enhancement of basic science and drug discovery would translate to broad societal benefit.This I-Corps project advances a new technology, supported by prototype data, to breach the major bottleneck in the rapidly growing cryo-EM field, with broad impact in life science and drug discovery. Cryo-EM is performed by taking images of frozen proteins using an electron microscope, then processing the images with a computer to obtain a 3D protein structure. To image and obtain a structure, the protein sample must first be frozen into a thin layer (~50-200 nm). With existing technology, this process is poorly controlled, lacks reproducibility and substantially adds to the time required to get a structure. Our device is designed to directly mitigate these sample preparation problems. It is a self-contained, freezable fluid cell nanofabricated to specification, giving full control over sample geometry and protein environment. Protein is passively loaded into the fluid cell, the loaded device is frozen, and can then be imaged directly. Materials on the top and bottom of the fluid cell are selected to transmit the electron beam through the protein sample with minimal noise. Successful prototype testing and data collection has shown the viability of the core design, with future work focused on refining the design and materials and streamlining manufacturing for eventual commercialization.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这个I-Corps项目的更广泛的影响和商业潜力如下：冷冻电子显微镜（Cryo-EM）是一种获得生物分子3D结构的方法。该方法在基础生命科学研究和制药部门中越来越重要，用于确定药物如何与其靶标相互作用。近年来，Cryo-EM作为一个整体已经成熟为一种强大的工具，并在2017年诺贝尔化学奖中达到顶峰，但用于3D研究的蛋白质“样品制备”过程并没有明显进展，并且被广泛认为是整个方法的主要瓶颈。这一瓶颈也带来了重大的商业机会。目前的项目推进了旨在解决这一瓶颈的基于纳米流体的新技术。这项技术的成功开发将产生一种可供商业化的消费品。这一目标的实现将大大推进和开辟新的可能性，在冷冻电镜结构测定。由此产生的基础科学和药物发现的加速和增强将转化为广泛的社会效益。这个I-Corps项目推进了一项新技术，由原型数据支持，以突破快速发展的cryo-EM领域的主要瓶颈，对生命科学和药物发现产生广泛影响。Cryo-EM是通过使用电子显微镜拍摄冷冻蛋白质的图像，然后用计算机处理图像以获得3D蛋白质结构来进行的。为了成像并获得结构，蛋白质样品必须首先冷冻成薄层（约50-200 nm）。利用现有技术，该过程控制不良，缺乏再现性，并且大大增加了获得结构所需的时间。我们的设备旨在直接缓解这些样品制备问题。它是一个独立的，可冷冻的流体细胞纳米制造的规格，提供完全控制样品的几何形状和蛋白质环境。蛋白质被动加载到流体细胞中，加载的设备被冷冻，然后可以直接成像。选择流体池顶部和底部的材料，以使电子束以最小的噪声穿过蛋白质样品。成功的原型测试和数据收集表明了核心设计的可行性，未来的工作重点是改进设计和材料，并简化制造，以最终实现商业化。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估来支持。