Genomics and Applied Biology 2024, Vol.15, No.1, 27-38 http://bioscipublisher.com/index.php/gab 35 collection and analysis possible, further improving the efficiency and resolution of structural biology research. Punjani et al. (2017) introduced cryoSPARC: a fast and unsupervised cryo EM structure determination algorithm that utilizes stochastic gradient descent (SGD) and branch and bound maximum likelihood optimization algorithms, enabling the main steps of cryo EM structure determination to be completed in a short period of time on affordable desktop computers. With the improvement of collection efficiency, the amount of data generated has also greatly increased, making data analysis a bottleneck in research. To address this challenge, researchers have developed a series of high-throughput data analysis tools that utilize advanced algorithms to automatically complete the process from image recognition to protein 3D structure reconstruction. The application of machine learning and artificial intelligence has further optimized this process, improved the speed and accuracy of data processing, and made the analysis of complex protein structures faster and more accurate. Kumar et al. (2021) introduced a user-friendly, high-throughput, and fully automated data acquisition software suitable for single particle cryo EM. By demonstrating the application of this software package in automatic imaging of SARS-CoV-2 spike proteins, the potential for fully automated image acquisition processes was emphasized. In addition, the integration of experimental processes is crucial for improving overall experimental efficiency. By integrating data collection, processing, and analysis into a continuous automated process, research efficiency has been significantly improved, while also ensuring standardization of the experimental process, reducing possible errors, and improving the reliability of structural analysis. This advancement in automation and integration not only accelerates the research of basic biology, but also has extremely important value in quickly understanding disease mechanisms, discovering new drug targets, and drug development. 4.3 Freezing electron microscopy combined with other technologies The combination of cryo electron microscopy (Cryo EM) technology and other scientific technologies is becoming an important trend in advancing the field of biomedical research. Through this interdisciplinary integration, scientists can analyze the structure and function of proteins from a more comprehensive and in-depth perspective, thereby deepening their understanding of the biomolecular world. For example, the combination of mass spectrometry technology not only enables researchers to accurately identify the composition and modification status of protein complexes, but also combines with spatial arrangement images provided by cryo electron microscopy to gain insight into complex biological processes at the molecular level. Engen and Komives (2020) emphasized the benefits of combining single particle cryo EM and hydrogen/deuterium exchange mass spectrometry (HDX-MS) methods, including low resolution density analysis, structural validation, analysis of individual proteins in large complexes, and research on isomers, protein quality control, and protein dynamics/motion during functional processes. By combining cryo electron microscopy with X-ray crystallography, a more accurate three-dimensional protein structure can be obtained based on the atomic level information provided by X-ray crystallography, while analyzing the structure of macromolecules using cryo electron microscopy. The application of this dual technology greatly improves the accuracy and reliability of structural analysis. Orlov et al. (2017) emphasized the integrated role of cryo EM in molecular and cellular structural biology, and examples of its combination with other methods such as X-ray crystallography, fluorescence imaging, or focused ion beam milling, such as studying ribosomes, viruses, chromatin, and nuclear receptors. Meanwhile, the combination of cryo electron microscopy and optical microscopy techniques, such as super-resolution microscopy and live cell imaging, provides possibilities for studying the function and behavior of proteins in a broader biological context. Li et al. (2018) introduced a new solution for non-integrated multiscale cryo related optical and electron microscopes (cryo CLEMs), enabling scientists to study biological samples at near atomic resolution while preserving their spatial localization in the cellular environment, aiming to improve
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