Japan Agency for Medical Research and Development (AMED)
the Cyclic Innovation for Clinical Empowerment (CiCLE)
日本
Japan Society for the Promotion of Science
24657111
日本
Japan Science and Technology
SENTAN
日本
引用
ジャーナル: J Struct Biol / 年: 2019 タイトル: A new cryo-EM system for single particle analysis. 著者: Tasuku Hamaguchi / Saori Maki-Yonekura / Hisashi Naitow / Yoshinori Matsuura / Tetsuya Ishikawa / Koji Yonekura / 要旨: A new cryo-EM system has been investigated for single particle analysis of protein structures. The system provides parallel illumination of a highly-coherent 300 kV electron beam from a cold-field ...A new cryo-EM system has been investigated for single particle analysis of protein structures. The system provides parallel illumination of a highly-coherent 300 kV electron beam from a cold-field emission gun, and boosts image contrast with an in-column energy filter and a hole-free phase plate. It includes motorized cryo-sample loading and automated liquid-nitrogen filling for cooling multiple samples. In this study, we describe gun and electron beam characteristics, and demonstrate the suitability of this system for single particle reconstructions. The performance of the system is tested on two examples, a spherical virus and apoferritin. GUI programs have also been developed to control and monitor the system for correct illumination, imaging with less ellipticity and steady magnification, and timing of flashing and liquid-nitrogen filling. These programs are especially useful for efficient application of the system to single particle cryo-EM.