With the rapid development of Cryo Electron Microscopy (Cryo EM) technology, significant progress has been made in protein structure analysis, opening up new avenues for research in the fields of biology and medicine. This study aims to review the latest progress of cryo electron microscopy technology in protein structure analysis, and explore its research significance and application prospects. Through in-depth analysis of technological optimization, method innovation, and its application in the study of complex biological macromolecular structures, it was found that cryoelectron microscopy technology not only improves resolution and signal-to-noise ratio, but also successfully analyzes various complex protein structures, providing powerful tools for research in the fields of biology and medicine. The development of cryoelectron microscopy technology not only deepens the understanding of protein structure and function relationships, but also provides new ideas and methods for drug development and disease treatment. Therefore, further promoting the development and application of cryoelectron microscopy technology is of great significance for promoting progress in the field of life sciences.