【摘要】 对生物大分子复合物的研究和结构分析对于全面了解其功能和生物学意义至关重要.冷冻电子显微镜在提供生物大分子结构及大分子分布等方面起到重要的作用.近年来,冷冻电子显微镜的硬件和软件的发展进一步提高了冷冻电子显微镜的有效性,使其对各种生物结构、蛋白质结构的解析更加准确快捷.但是,对于生物系统来说,蛋白质和大分子复合物等均处于复杂的生理环境中,因此原位检测生物分子的三维结构对于生物体系和结构生物学具有重要意义.冷冻电子断层扫描作为一种功能强大的技术,可以无需标记直接通过冷冻样品的固有衬度识别生物大分子的结构,并且可在原位生理环境中对生物分子进行纳米级分辨率的三维成像.本文综述了与冷冻电子断层扫描相关的样品制备和数据处理技术,并总结了冷冻电子断层扫描技术在分离的大分子复合物和整个细胞或组织中的生物学应用.更多还原

【Abstract】 The study and structural analysis of biological macromolecular complexes are crucial for a comprehensive understanding of their function and biological significance. The cryo-electron microscopy has become the mainstream technology for providing information about the structure and distribution of macromolecules in biology. In addition,the advancement of hardware and software in rencet years has further improved the effectiveness of cryo-electron microscopy,making it more accurate and faster to analyze various biological structures and proteins. However,for biological systems,proteins or macromolecular complexes are in a complex physiological environment,so in situ detection of the 3 D biological structures is crucial for biological system and structural biology. As a powerful technology,cryo-electron tomography can rely on the intrinsic contrast of frozen samples to identify the structure of biological macromolecules directly without marking,and can perform 3 D imaging with nano-resolution in their native environment. In this review,we summerized the methods of sample preparation and technical aspects relevant for cryo-electron tomography as well as the biological applications from isolated macromolecular complexes to entire cells and tissues.更多还原