School of Medicine, Shaoguan University, Shaoguan, Guangdong, China.
Kobilka Institute of Innovative Drug Discovery, School of Medicine, the Chinese University of Hong Kong, Shenzhen, Guangdong, China.
Methods Mol Biol. 2025;2854:177-188. doi: 10.1007/978-1-0716-4108-8_18.
Cryo-electron microscopy is a powerful methodology in structural biology and has been broadly used in high-resolution structure determination for challenging samples, which are not readily available for traditional techniques. In particular, the strength of super macro-complexes and the lack of a need for crystals for cryo-EM make this technique feasible for the structural study of complexes involved in antiviral innate immunity. This chapter presents detailed information and experimental procedures of Cryo-EM for determining the structures of the complexes using STING as an example. The procedures included a sample quality check, high-resolution data acquisition, and image processing for Cryo-EM 3D structure determination.
冷冻电子显微镜技术是结构生物学领域的一种强大方法，已广泛应用于难以用传统技术获得的高分辨率结构测定的挑战性样品中。特别是，超级大分子复合物的优势以及冷冻电镜不需要晶体的特点，使得该技术可用于抗病毒先天免疫复合物的结构研究。本章以 STING 为例，介绍了用于确定复合物结构的冷冻电镜的详细信息和实验步骤。这些步骤包括样品质量检查、高分辨率数据采集和冷冻电镜 3D 结构测定的图像处理。
