Abstract
A cryo-electron microscopy (cryo-EM) system equipped with a cold-field emission gun and an energy filter is well-suited for higher-resolution single particle analysis and high-precision 3D electron crystallography. We designed such system and installed a JEOL CRYO ARM 300 electron microscope- the first machine of this model. The microscope has provided high quality data in both imaging and diffraction since July 2018. For single particle analysis, we are routinely able to reconstruct higher-resolution structures with better estimated B-factors for map sharpening from smaller numbers of images than can be obtained with conventional Schottky emission. For 3D electron crystallography, we are also routinely able to obtain highest-quality diffraction data thanks to higher-energy electrons and energy filtration. Automated data collection allows high-throughput structure determination in both applications. This review introduces our recent results, progress, and perspective for running such cryo-EM system.
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Acknowledgements
We thank the JEOL cryo-EM group for technical assistance in setting up the CRYO ARM 300 electron microscope, Yasuhiro Kashino for collaborative work on A. marina photosystem I, NobuyukiYoshikawa for providing the ALSV sample, Haruaki Yanagisawa for providing a plasmid of mouse apoferritin, Satoru Inoue, Tatsuo Hasegawa, Yasutomo Segawa for providing crystal samples of organic molecules. The works presented in this review were partly supported by Japan Society for the Promotion of Science Grant-in-Aid for Scientific Research Grant 16H04757 (to K.Y.), Japan Society for the Promotion of Science Grant-in-Aid for Challenging Exploratory Research Grant 24657111 (to K.Y.), the RIKEN Pioneering Project, Dynamic Structural Biology (to K.Y., S.M.-Y., T.H., H.N. and K.T.), the Cyclic Innovation for Clinical Empowerment (CiCLE) from the Japan Agency for Medical Research and Development, AMED(to K.Y.), JST CREST Grant Number JPMJCR18J2, Japan (to K.Y.), and JST-Mirai Program Grant Number JPMJMI20G5 (to K.Y.).
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Yonekura, K., Maki-Yonekura, S., Hamaguchi, T., Naitow, H., Takaba, K., Kawakami, K. (2021). Cryo-EM and ED with a Cold-Field Emission Beam and Energy Filtration. In: Ghosal, P., Carter, C.B., Vinothkumar, K.R., Sarkar, R. (eds) Applications of Microscopy in Materials and Life Sciences. Springer Proceedings in Materials, vol 11. Springer, Singapore. https://doi.org/10.1007/978-981-16-2982-2_23
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