Table 2. Cryo-EM data collection and processing.
| Dataset 1 | Dataset 2 | Dataset 3 | Dataset 4 | Dataset 5 | |
|---|---|---|---|---|---|
| Microscope | Titan Krios (m06 eBIC) | Titan Krios (m06 eBIC) | Titan Krios (m06 eBIC) | Titan Krios (MRC-LMB) |
Titan Krios (MRC-LMB) |
| Pixel Size | 1.380 | 1.380 | 1.380 | 1.179 | 1.390 |
| Voltage | 300 | 300 | 300 | 300 | 300 |
| Spherical Aberation | 2.7 | 2.7 | 2.7 | 2.7 | 2.7 |
| Total exposure (e-/Å2) | 39.60 | 42.50 | 37.77 | 39.36 | 44.36 |
| Exposure Length (s) | 5.0 | 11.02 | 14 | 11 | 11 |
| Frames | 25 | 44 | 40 | 44 | 40 |
| Defocus Range (µm) | −0.5 to −1.5 | −0.5 to −1.5 | −0.5 to −1.5 | −0.5 to −1.5 | −0.5 to −1.5 |
| Micrographs | 2776 | 2484 | 1206 | 4228 | 932 |
| Microscope tilt (degrees) | 0 | 0 | 30 | 20 | 20 |
| Volta Phase Plate | ✓ | ✓ | ✓ | ✓ | ✓ |
| Pre-merge processing | Dataset 1 | Dataset 2 | Dataset 3 | Dataset 4 | Dataset 5 |
| Motion Correction and CTF estimation (micrographs) | 2776 | 2484 | 1206 | 4228 | 932 |
| Blob-based autopicking (particles) | 1,298,488 | 541,233 | 319,989 | 1,145,157 | 223,613 |
| 2x iterations of 2D classification (particles) | 63,180 | 63,315 | 51,371 | 305,685 | 28,662 |
| Template based autopicking (particles) | 467,323 | 687,645 | 826,006 | N/A | 273,852 |
| 2D classification (particles) | 11,862 | 71,977 | 63,881 | 113,852 | 66,799 |
| Ab Initio 3D classification (particles) | 103,105 | 71,977 | 46,349 | 113,852 | 50,232 |
| Post-merge processing | Combined Datasets | ||||
| 2x Iterations ab initio 3D classification (particles) | 405,515 | ||||
| Non-uniform Refinement (particles) | 167,294 | ||||
| Per-particle CTF refinement | 6.71 Å map | ||||
| Non-uniform refinement with local resolution estimation and filtering | 6.4 Å map | ||||
| EMDB Deposition code | EMD-11058 | ||||