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Cryo-electron microscopy (cryo-EM), especially single-particle analysis, has become a powerful technique for visualizing the structure of biological macromolecules at high resolution. However, data acquisition in cryo-EM is time consuming because it requires the collection of thousands of images to achieve a high-quality reconstruction. Real-time preprocessing and display of the images can greatly enhance the efficiency and quality of data collection. This study presents AutoEMage, a new open-source software package that automates data transfer, preprocessing and real-time display in cryo-EM experiments. AutoEMage also includes a real-time data monitoring system that alerts users to issues with their data, allowing them to take corrective actions accordingly. The software is equipped with an easy-to-use graphical user interface that provides seamless data screening and real-time feedback on data quality and microscope status.

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