REL
2020-05-13
2021-05-17
2021-05-17
Single-Particle CryoEM of Human Apoferritin Light Chain Vitrified Using Back-it-up
0000-0001-6656-6320
Yong Zi
Tan
The Hospital for Sick Children
686 Bay Street
Toronto
Ontario
Canada
M5G0A4
0000-0003-0566-2209
John
L
Rubinstein
The Hospital for Sick Children
686 Bay Street
Toronto
Ontario
Canada
M5G0A4
Tan YZ
Rubinstein JL
2.1
10.6019/EMPIAR-10421
EMDB
EMD-21951
Tan YZ
Rubinstein JL
Through-grid wicking enables high-speed cryoEM specimen preparation
bioRxiv
2020
10.1101/2020.05.03.075366
Tan YZ
Rubinstein JL
Through-grid wicking enables high-speed cryoEM specimen preparation
Acta crystallographica. Section D, Structural biology
Acta Crystallogr D Struct Biol
Pt 11
76
1092
1103
2020
English
10.1107/s2059798320012474
33135680
Unaligned Falcon IV movie frames
/data/01_Raw_Frames
micrographs - multiframe
MRCS
MRCS
3168
30
1
30
SIGNED 16 BIT INTEGER
4096
0.816
4096
0.816
3,168 movies were acquired from two grids of apoferritin prepared with BIU. The pixel size was calibrated at 0.816 Å/pixel by fitting a crystal structure of human ferritin L chain (PDB ID: 2FFX) into a preliminary 3D map. Movies were collected for 9 sec with 29 exposure fractions, a camera exposure rate of ~3 e-/pix/sec, and total specimen exposure of ~40 e-/Å^2. Note: Due to software issues, the last frames in these raw movies have errors. Please exclude the last frame (frame 30) when doing any alignments.
data/02_Aligned_Micrographs/*mrc
data/03_Final_Particle_Stack/BIU_HuApo_Final.star
data/03_Final_Particle_Stack
Aligned Dose-Weighted Micrographs
/data/02_Aligned_Micrographs
micrographs - single frame
MRC
MRC
3168
1
32 BIT FLOAT
4096
0.816
4096
0.816
Exposure fractions were aligned with MotionCor2 within Relion, using 7x7 patches and a B factor of 500 pixel^2 for global alignment and 150 pixel^2 for local alignment. There is a micrographs_ctf.star file with CTF estimation from CTFFind4. Note: Due to software issues, the last frames in these raw movies have errors. Please exclude the last frame (frame 30) when doing any alignments.
data/02_Aligned_Micrographs/*mrc
data/03_Final_Particle_Stack/BIU_HuApo_Final.star
data/03_Final_Particle_Stack
Final Particle Stack with Final Euler Angles and Shifts
/data/03_Final_Particle_Stack
picked particles - multiframe - processed
MRCS
MRCS
1
594259
1
594259
32 BIT FLOAT
288
0.816
288
0.816
Final polished particle stack with Euler angles and shifts. Higher order CTF aberration corrections were done within CryoSPARC 2 but the resulting parameters were not able to be exported to .star format here. When using this particle stack, it is recommended to do higher order CTF aberration corrections again in CryoSPARC/Relion to obtain the best resolution.
data/02_Aligned_Micrographs/*mrc
data/03_Final_Particle_Stack/BIU_HuApo_Final.star
data/03_Final_Particle_Stack