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

这个i-Corps项目的更广泛的影响和商业潜力如下：低温电子显微镜(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部门确定药物如何与其目标相互作用方面具有越来越重要的作用。近年来，冷冻-EM作为一个整体已经成熟为一个强大的工具，最终获得了2017年的诺贝尔化学奖，但用于3D研究的蛋白质样品准备过程并没有取得显著进展，人们普遍认为这是整个方法的主要瓶颈。这一瓶颈也带来了一个重大的商业机会。本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生产出一种随时可以商业化的消费品。这一目标的实现将极大地推进和开启低温EM结构确定的新可能性。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，以突破快速增长的低温电磁领域的主要瓶颈，对生命科学和药物发现产生广泛影响。冷冻-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来获得3D蛋白质结构来进行的。为了成像和获得结构，蛋白质样品必须首先被冷冻成一层薄的(~50-200 nm)。在现有技术下，这一过程控制不佳，缺乏重复性，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。它是一种自给自足的、可冷冻的流体细胞，符合规格，可以完全控制样品的几何形状和蛋白质环境。蛋白质被被动地加载到流体细胞中，加载的设备被冻结，然后可以直接成像。流体池顶部和底部的材料被选择来以最小的噪声通过蛋白质样品来传输电子束。成功的原型测试和数据收集表明了核心设计的可行性，未来的工作重点是完善设计和材料，并为最终商业化而简化制造。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.