PDF(Pubmed)
Abstract：
Single-particle cryo-electron microscopy has become a widely adopted method in structural biology due to many recent technological advances in microscopes, detectors and image processing. Before being able to inspect a biological sample in an electron microscope, it needs to be deposited in a thin layer on a grid and rapidly frozen. The VitroJet was designed with this aim, as well as avoiding the delicate manual handling and transfer steps that occur during the conventional grid-preparation process. Since its creation, numerous technical developments have resulted in a device that is now widely utilized in multiple laboratories worldwide. It features plasma treatment, low-volume sample deposition through pin printing, optical ice-thickness measurement and cryofixation of pre-clipped Autogrids through jet vitrification. This paper presents recent technical improvements to the VitroJet and the benefits that it brings to the cryo-EM workflow. A wide variety of applications are shown: membrane proteins, nucleosomes, fatty-acid synthase, Tobacco mosaic virus, lipid nanoparticles, tick-borne encephalitis viruses and bacteriophages. These case studies illustrate the advancement of the VitroJet into an instrument that enables accurate control and reproducibility, demonstrating its suitability for time-efficient cryo-EM structure determination.
摘要：
由于显微镜的许多最新技术进步，单粒子低温电子显微镜已成为结构生物学中广泛采用的方法。探测器和图像处理。在能够在电子显微镜中检查生物样本之前，它需要沉积在网格上的薄层中并迅速冷冻。VitroJet就是为了这个目的而设计的，以及避免在常规网格制备过程中发生的精细手动处理和转移步骤。自创建以来，许多技术的发展已经导致了现在在全球多个实验室中广泛使用的设备。它的特点是等离子治疗，通过针脚印刷进行少量样品沉积，通过喷射玻璃化对预剪的Autogrids进行光学冰厚测量和冷冻固定。本文介绍了VitroJet的最新技术改进以及它为cryo-EM工作流程带来的好处。显示了各种各样的应用：膜蛋白，核小体,脂肪酸合成酶，烟草花叶病毒,脂质纳米粒，蜱传脑炎病毒和噬菌体。这些案例研究说明了将VitroJet推进到能够实现精确控制和可重复性的仪器中，证明其适用于时间有效的低温EM结构测定。
