Krios G4 apoferritin test with Falcon 4 (no filter) EPU 1x1x4 [3540 multi-frame micrographs composed of 720 frames each in EER format]

Cryo-EM Performance Testing of Hardware and Data Acquisition Strategies

Danev R, Yanagisawa H, Kikkawa M

Microscopy (Oxford, England) 1 (2021) 1-2

PMID: 33969878

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  • micrographs - multiframe
1. Unaligned multi-frame non-gain-normalized EER movies in TIFF format
micrographs - multiframe
Image format:
No. of images or tilt series:
Image size:
(4096, 4096)
Pixel type:
Pixel spacing:
(0.3948, 0.3948)
Please check the "dataset_details_apoF_EPU.txt" for details.

To test the overall performance of the Krios G4 at The University of Tokyo, five datasets were collected from the same apoferritin grid in the beginning of July 2020.
This was the third dataset in the series.

Sample: mouse heavy chain apoferritin
Grid: UltrAuFoil 300 mesh R1.2/1.3
Cemera: Falcon 4 installed at pre-fitler camera position
Camera mode: EER
TEM mode: NanoProbe
Mag: x195k
C2 aperture: 50 um
Spot: 6
Beam diameter: 0.65 um
Data collection software: EPU
Acquisition method: beam-image shift without beam-tilt compensation
Acquisition pattern: 1 x 1 holes, 4 images around the periphery of the hole
Physical Pixel: 0.3948 A/pix
EER 2x oversamples pixel: 0.1974 A/pix
Exposure rate: 2.74 e/pix/s (measured in EPU)
Exposure time: 3.0 s
Total exposure: 52.7 e/A2
Frames: 720 EER frames, processed with 10 frames/fraction resulting in 72 fractions
Defocus: 0.2 - 1.0 um
Movies: 3540
Data format: non-gain-normalized EER encapsulated in TIFF format
Gain reference: "gain_2xUps_8k.mrc" prepared with the command "relion_estimate_gain --i Import/job001/movies.star --o gain_2xUps_8k.mrc --eer_upsampling 2 --dont_invert --j 30" from the data
Final particles: 135k
Particles file: "run_data.star"
Optics groups: per-acquisition position and per hour
Resolution (with Ewald correction): 1.43 A
Rosenthal-Henderson B-factor: 42.9 A^2

November 24, 2020

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