A high precision piezo driven replacement goniometer for cryoelectron microscopy

项目摘要 项目名称：一种高精度的压电式低温电子替代测角仪 显微镜 公司名称：dba蜂鸟精密机械有限公司 主要调查者：约瑟夫·史蒂维克 摘要： 用低温电子显微镜(Cryo-EM)解析的生物分子结构 大大提高了我们对生命过程的理解，特别是在以下领域 药物设计、疫苗和其他微生物健康解决方案。这项技术具有 它是如此的成功和具有变革性，以至于它被授予诺贝尔化学奖 2017年，以及最近被用来解决依赖RNA的RNA的结构 新冠肺炎病毒的聚合酶。Cryo-EM生产的生化模型 像这样的研究对研究抗病毒药物和其他微生物学是至关重要的 解决方案除了帮助我们形成对分子结构的基本理解之外。 然而，在Cryo-EM硬件方面存在着显著的技术差距 被忽视了。透射电子显微镜侧入测角仪(样品 定位系统)是在开发低温电磁技术之前设计的， 低温侧向支撑架不稳定，使用不便。因此，闭环系统 自动化的Cryo-EM工作流变得效率低下，并且受限于什么定位 系统可以实现什么，而不是科学上重要的东西。耗时的图像 目前使用跟踪和纠错方法来补偿穷人 测角器和侧孔支架的机械定位能力。 我们的解决方案是设计一个现代化的样机运动控制台和相应的 样品架作为专门构建的组合，针对重复性、准确性、 以及在低温下的稳定性。拟议的设计将改善现有的 ART Cryo-EM自动化在精度和 将杜瓦寿命提高5倍。这一改进是由一种独特的 可同时优化工作台、托架和低温系统的地面设计。在……里面 目标1，我们将优化舞台和支架，利用一种非传统的设计，限制 耦合运动，并采用现代机电技术和传感方法 这将极大地提高我们舞台的闭环精度 最新的测角仪。在目标2中，我们将优化低温系统设计，以实现-170°C的稳定 样品温度和易用性与舞台和支架相协调。作为一名 商业产品，该设备将作为测角器的简单改装替代品 在已经致力于生物研究的数千个TEM上。我们的长期合作 目标是让更多的科学家拥有他们需要的仪器来实现 无需解决基本硬件问题即可获得最佳的Cryo-EM结果。

Project Summary Project Title: A high precision piezo driven replacement goniometer for cryoelectron microscopy Company Name: Hummingbird Precision Machine Co., dba Hummingbird Scientific Principal Investigator: Joseph Stevick Summary: Biomolecular structures resolved by cryoelectron microscopy (Cryo-EM) have significantly advanced our understanding of life processes, specifically in the areas of drug design, vaccines, and other microbiological health solutions. The technique has been so successful and transformative that it was awarded the Nobel Prize for Chemistry in 2017, and more recently was used to solve the structure of the RNA-dependent RNA polymerase from the COVID-19 virus. The biochemical models produced by Cryo-EM studies like this one are vital to researching anti-viral drugs and other microbiological solutions in addition to helping shape our basic understanding of molecular machinery. However, there is a significant technology gap in Cryo-EM hardware that has been overlooked. Transmission electron microscope (TEM) side-entry goniometers (sample positioning systems) were designed before the Cryo-EM technique was developed, and cryogenic side-entry holders are unstable and inconvenient. As a result, closed-loop automated of Cryo-EM workflows becomes inefficient and limited to what the positioning system can achieve rather than what is scientifically important. Time consuming image tracking and error correction methods are currently used to compensate for the poor mechanical positioning capabilities of goniometers and side-entry holders. Our solution is to design a modern sample motion control stage and a corresponding sample holder as a purpose-built combination that is optimized for repeatability, accuracy, and stability at cryogen temperatures. The proposed design will improve state-of-the- art Cryo-EM automation by more than an order of magnitude in precision and increase Dewar life by a factor of 5. This improvement is made possible by a unique ground-up design that optimizes the stage, holder, and cryo-system simultaneously. In Aim 1, we will optimize the stage and holder, utilizing an unconventional design that limits coupled motion, and employs modern mechatronic technologies and sensing methods that will dramatically improve the closed-loop precision of our stage in comparison with current goniometers. In Aim 2, we will optimize the cryo-system design for stable -170°C sample temperatures and ease of use in concert with the stage and holder. As a commercial product, this device would act as a simple retrofit replacement to goniometers on thousands of TEMs that are already dedicated to biological research. Our long-term objective is to empower more scientists with the instruments they need to achieve the best possible Cryo-EM results without having to struggle with basic hardware issues.