A high precision piezo driven replacement goniometer for cryoelectron microscopy

项目摘要 项目名称：一种用于低温电子的高精度压电驱动替代测角仪 显微镜 公司名称：蜂鸟精密机械有限公司dba蜂鸟科技 主要研究者：Joseph Stevick 总结： 通过冷冻电子显微镜（Cryo-EM）解析的生物分子结构 极大地推进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。该技术具有 它是如此的成功和变革，以至于它被授予诺贝尔化学奖， 在2017年，最近被用来解决依赖RNA的RNA的结构， COVID-19病毒的聚合酶。Cryo-EM产生的生化模型 像这样的研究对于研究抗病毒药物和其他微生物制剂至关重要。 解决方案，除了帮助塑造我们对分子机制的基本理解。 然而，在冷冻EM硬件方面存在重大技术差距， 忽视透射电子显微镜（TEM）侧入式测角仪（样品 定位系统）是在冷冻EM技术开发之前设计的， 低温侧入式支架不稳定且不方便。因此，闭环 冷冻EM工作流程的自动化变得效率低下，并且局限于定位 而不是科学上重要的东西。耗时图像 跟踪和误差校正方法目前用于补偿 测角器和侧入式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品运动控制平台和相应的 样品保持器作为一个专用的组合，优化了重复性，准确性， 和在冷冻剂温度下的稳定性。设计方案将改善... 先进的Cryo-EM自动化技术在精度上提高了一个数量级， 杜瓦瓶寿命提高5倍。这种改进是由一种独特的 全新设计，同时优化载物台、保持器和冷冻系统。在 目标1，我们将优化载物台和保持器，利用非传统的设计， 耦合运动，并采用现代机电技术和传感方法 这将大大提高我们的阶段的闭环精度相比， 电流测角仪在目标2中，我们将优化低温系统设计，以实现稳定的-170 ° C 样品温度和与载物台和保持器配合使用的易用性。作为 商业产品，该设备将作为一个简单的改造替代量角器 已经致力于生物学研究的数千个TEM上。我们的长期 我们的目标是让更多的科学家拥有他们所需要的工具， 最好的冷冻EM结果，而不必与基本的硬件问题作斗争。

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.