|Entry||Database: EMDB / ID: EMD-9890|
|Title||Cryo-EM structure of human apoferritin at 1.9A resolution by CRYO ARM 300|
|Source||Homo sapiens (human)|
|Method||single particle reconstruction / cryo EM / Resolution: 1.9 Å|
|Authors||Hamaguchi T / Maki-Yonekura S / Naitow H / Matsuura Y / Ishikawa T / Yonekura K|
|Citation||Journal: J. Struct. Biol. / Year: 2019|
Title: A new cryo-EM system for single particle analysis.
Authors: Tasuku Hamaguchi / Saori Maki-Yonekura / Hisashi Naitow / Yoshinori Matsuura / Tetsuya Ishikawa / Koji Yonekura /
Abstract: 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.
|Date||Deposition: Apr 19, 2019 / Header (metadata) release: May 15, 2019 / Map release: May 15, 2019 / Update: May 15, 2019|
|Structure viewer||EM map: |
Downloads & links
|File||Download / File: emd_9890.map.gz / Format: CCP4 / Size: 512 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
|Voxel size||X=Y=Z: 0.495 Å|
|Symmetry||Space group: 1|
CCP4 map header:
|Entire||Name: apoferritinFerritin / Number of components: 1|
-Component #1: cellular-component, apoferritin
|Cellular-component||Name: apoferritinFerritin / Recombinant expression: No|
|Source||Species: Homo sapiens (human)|
|Specimen||Specimen state: Particle / Method: cryo EM|
|Sample solution||Specimen conc.: 3.5 mg/mL / pH: 7.5|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: ETHANE / Temperature: 277 K / Humidity: 100 %|
-Electron microscopy imaging
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 101 e/Å2 / Illumination mode: SPOT SCAN|
|Lens||Magnification: 100000.0 X (nominal) / Cs: 2.7 mm / Imaging mode: BRIGHT FIELD / Defocus: 3000.0 - 13000.0 nm|
|Specimen Holder||Model: JEOL|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Image acquisition||Number of digital images: 1292|
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