Table 1 |.
CryoET data collection and processing details
| Titan Krios† | GagT8I | Gag WT (EMPIAR-10164) | Ribosome (EMPIAR-10304) | Apoferritin |
|---|---|---|---|---|
|
| ||||
| Voltage (kV) | 300 | 300 | 300 | 300 |
| Detector | Falcon 4 | Gatan K2 | Gatan K3 | Gatan K3 |
| Energy filter | Selectris X, 10 eV slit | Gatan bioquantum, 20 eV | Gatan bioquantum, 20 eV | Gatan bioquantum, 20 eV |
| Super-resolution mode | Yes | Yes | Yes | Yes |
| Pixel size (Å) | 1.18 | 1.35 | 2.1 | 1.34 |
| Total electron dose (e-/Å2) | 122 | ~120 | ~120 | 102 |
| Dose rate (e-/Å2/s ) | 3 | 3 | 4.2 | |
| Frame number | 10 | 10 | 10 | |
| Acquisition scheme | −60/60°, 3° | −60/60°, 3° | −60/60°, 3° | −60/60°, 3° |
| Defocus range (μm) | −1.36 ~ −3.11 | −1.5 ~ −3.96 | −2.2 ~ −4.3 | −1.5 ~ −3.5 |
| Number of tilt-series | 5 | 5 | 12 | 6 |
| Software | IMOD, emClarity | IMOD, emClarity | IMOD, emClarity | IMOD, emClarity |
| Number of tomograms | 5 | 5 | 12 | 6 |
| Number of initial subtomograms | 20010 | 15791 | 10441 | 5668 |
| Number of subtomograms after classification | 13844 | 15460 | 8131 | 4826 |
| Symmetry imposed | C6 | C6 | C1 | O |
| Resolution at 0.143 FSC | 5.0 Å/4.5 Å* | 3.3 | 7.0 | 2.8 |
| Data deposited | EMPIAR-10643 EMD-13390 | EMD-13354 | EMD-13270 | EMPIAR-10787 EMD-13271 |
†
Titan Krios is a 300kV electron microscope used for cryoEM data collection: https://www.thermofisher.com/uk/en/home/electron-microscopy/products/transmission-electronmicroscopes/krios-g4-cryo-tem.html
*
Density maps were calculated using subtomograms averaged in emClarity (5.0 Å) or using projection images (4.5 Å) reconstructed by cisTEM implemented in emClarity.