|Entry||Database: EMDB / ID: EMD-9261|
|Title||Electron cryotomogram of Bdellovibrio bacteriovorus acquired by continuous tilting|
|Sample||Bdellovibrio bacteriovorus (bacteria)|
|Biological species||Bdellovibrio bacteriovorus (bacteria)|
|Method||electron tomography / cryo EM|
|Authors||Chreifi G / Chen S / Metskas LA / Kaplan M / Jensen GJ|
|Citation||Journal: J. Struct. Biol. / Year: 2019|
Title: Rapid tilt-series acquisition for electron cryotomography.
Authors: Georges Chreifi / Songye Chen / Lauren Ann Metskas / Mohammed Kaplan / Grant J Jensen /
Abstract: Using a new Titan Krios stage equipped with a single-axis holder, we developed two methods to accelerate the collection of tilt-series. We demonstrate a continuous-tilting method that can record a ...Using a new Titan Krios stage equipped with a single-axis holder, we developed two methods to accelerate the collection of tilt-series. We demonstrate a continuous-tilting method that can record a tilt-series in seconds, but with loss of details finer than ∼4 nm. We also demonstrate a fast-incremental method that can record a tilt-series several-fold faster than current methods and with similar resolution. We characterize the utility of both methods in real biological electron cryotomography workflows. We identify opportunities for further improvements in hardware and software and speculate on the impact such advances could have on structural biology.
Downloads & links
|File||Download / File: emd_9261.map.gz / Format: CCP4 / Size: 1019.5 MB / Type: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)|
|Projections & slices|
Images are generated by Spider.
generated in cubic-lattice coordinate
|Voxel size||X=Y=Z: 17.28 Å|
|Symmetry||Space group: 1|
CCP4 map header:
-Entire Bdellovibrio bacteriovorus
|Entire||Name: Bdellovibrio bacteriovorus (bacteria) / Number of components: 1|
-Component #1: cellular-component, Bdellovibrio bacteriovorus
|Cellular-component||Name: Bdellovibrio bacteriovorus / Recombinant expression: No|
|Source||Species: Bdellovibrio bacteriovorus (bacteria)|
|Specimen||Specimen state: Cell / Method: cryo EM|
|Sample solution||pH: 7.8|
|Vitrification||Instrument: FEI VITROBOT MARK IV / Cryogen name: OTHER / Temperature: 296 K / Humidity: 100 %|
-Electron microscopy imaging
Model: Titan Krios / Image courtesy: FEI Company
|Imaging||Microscope: FEI TITAN KRIOS|
|Electron gun||Electron source: FIELD EMISSION GUN / Accelerating voltage: 300 kV / Electron dose: 0.08 e/Å2 / Illumination mode: FLOOD BEAM|
|Lens||Imaging mode: BRIGHT FIELD|
|Specimen Holder||Model: OTHER|
|Camera||Detector: GATAN K2 SUMMIT (4k x 4k)|
|Processing||Method: electron tomography / Number of sections: 1260|
|3D reconstruction||Algorithm: SIMULTANEOUS ITERATIVE (SIRT) / Software: TOMO3D|
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