Krios G4 apoferritin test with K3/(slit out) SerialEM BIS 1x1x4 [4184 multi-frame micrographs composed of 76 frames each in TIFF 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 movies in LZW compressed TIFF format
micrographs - multiframe
Image format:
No. of images or tilt series:
Image size:
(11520, 8184)
Pixel type:
Pixel spacing:
(0.1997, 0.1997)
Please see the "dataset_details_apoF_set3_noEF.txt" file 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 last, fifth dataset in the series.

Sample: mouse heavy chain apoferritin
Grid: UltrAuFoil 300 mesh R1.2/1.3
Cemera: Gatan K3 post a BioQuantum energy filter
Camera mode: super resolution, CDS
TEM mode: EFTEM NanoProbe
Mag: x215k
C2 aperture: 50 um
Spot: 6
Beam diameter: 0.65 um
EF slit: out
Data collection software: SerialEM
Acquisition method: beam-image shift with beam-tilt compensation
Acquisition pattern: 1 x 1 holes, 4 images around the periphery of the hole
Physical Pixel: 0.3994 A/pix
SuperRes pixel: 0.1997 A/pix
Exposure rate: 3.62 e/pix/s (CDS)
Exposure time: 3.04 s
Total exposure: 69.0 e/A2
Frames: 76
Defocus: 0.2 - 0.9 um
Movies: 4184
Data format: non-gain-normalized LZW compressed TIFFs
Gain reference: "apoF_set3_noEF_gain.mrc" prepared from the data
Final particles: 113k
Particles file: "run_data.star"
Optics groups: per-acquisition position
Resolution (with Ewald correction): 1.43 A
Rosenthal-Henderson B-factor: 42.7 A^2
Note: The air-conditioning temperature of the Krios G4 room was reduced by 2 degC around noon on the second day (July 13th) which caused a gradual increase in beam tilt/coma in the data. Therefore, only data from the first two acquired grid squares (314 and 321) was used.

November 26, 2020

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