研究生物大分子机器的结构和功能是理解细胞内生命过程的关键要素之一，近年来随着直接电子探测的成功应用和图像分析处理算法的飞速发展，冷冻电镜单颗粒分析技术给生物大分子机器的结构分析带来了革命性的进步，未来在细胞原位直接解析特定生物大分子机器的三维结构并分析其在细胞内部的结构动态将成为可能。本项目将通过发展细胞/组织样品冷冻减薄、冷冻光电关联成像、高分辨率冷冻电子断层数据收集和三维重构以及三维体数据精确均类等技术建立生物大分子机器的细胞原位结构解析和动态分析技术，并实现对典型超大分子复合体的亚纳米分辨率原位结构解析。

Understanding the structure and function of biological macromolecular machinery is one of the keys to resolve the mystery of life in the cell. With the recent success of direct electron detector and sophisticated image processing algorithms, the technique of cryo-electron microscopy single particle analysis has brought a revolution of structural biology to the structural analysis of macromolecular machineries. In the future, solving the structures of those machineries directly from their cellular context and analyzing their structural dynamics in situ will become possible. Here, we aim to develop a novel technology for in situ structural analysis of biological macromolecular machineries by developing cell/tissue cryo-thinning, cryo-fluorescence and electron microscopy, data collection, 3D reconstruction, and sub-tomogram classification and averaging techniques. We will also select a couple of importance macromolecular machineries such as RyR1 to give a proof-of-concept of our technology by obtaining their structures in-situ with sub-nanometer resolution.