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

该I-Corps项目的广泛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三维结构的方法。该方法在基础生命科学研究和制药领域越来越重要，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”过程并没有明显的进步，被广泛认为是整个方法的主要瓶颈。这个瓶颈也提供了一个重要的商业机会。目前的项目推进了新的纳米流体技术，旨在解决这一瓶颈。这项技术的成功开发将产生可用于商业化的可消费产品。这一目标的实现将极大地推进和打开冷冻电镜结构测定的新可能性。由此产生的基础科学和药物发现的加速和加强将转化为广泛的社会效益。这个I-Corps项目在原型数据的支持下，推进了一项新技术，突破了快速发展的低温电镜领域的主要瓶颈，在生命科学和药物发现方面具有广泛的影响。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蛋白质结构来执行的。为了成像并获得结构，必须首先将蛋白质样品冷冻成薄层（~50- 200nm）。利用现有技术，这一过程控制不佳，缺乏可重复性，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规格，给予样品几何形状和蛋白质环境的完全控制。蛋白质被被动地装载到流体细胞中，装载的设备被冷冻，然后可以直接成像。选择液体电池顶部和底部的材料，以最小的噪声传输电子束通过蛋白质样品。成功的原型测试和数据收集表明了核心设计的可行性，未来的工作将集中在改进设计和材料以及简化生产以最终实现商业化。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

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.