Cryo-electron microscopy (cryo-EM) is a structural biological method that is used to determine the 3D structures of biomacromolecules. After years of development, cryo-EM has made great achievements, which has led to a revolution in structural biology. In this article, the principle, characteristics, history, current situation, workflow, and common problems of cryo-EM are systematically reviewed. In addition, the new development direction of cryo-EM-cryo-electron tomography (cryo-ET), is discussed in detail. Also, cryo-EM is prospected from the following aspects: the structural analysis of small proteins, the improvement of resolution and efficiency, and the relationship between cryo-EM and drug development. This review is dedicated to giving readers a comprehensive understanding of the development and application of cryo-EM, and to bringing them new insights.

中文翻译：

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低温电子显微镜的发展，应用和前景。

低温电子显微镜（cryo-EM）是一种结构生物学方法，用于确定生物大分子的3D结构。经过多年的发展，cryo-EM取得了举世瞩目的成就，从而引发了结构生物学的一场革命。在本文中，系统地回顾了cryo-EM的原理，特性，历史，现状，工作流程和常见问题。此外，详细讨论了低温-EM-Cryo-Electro层析成像（cryo-ET）的新发展方向。另外，可从以下几个方面对cryo-EM进行展望：小蛋白的结构分析，分辨率和效率的提高以及cryo-EM与药物开发之间的关系。这篇评论致力于使读者全面了解cryo-EM的开发和应用，