Table 1 In situ cryo-FIB-ET data collection and reconstruction statistics
Labeling the ribosome | Labeling H2AFY | |||||
|---|---|---|---|---|---|---|
In situ labeled 5-nm-HN | In situ labeled 1.4-nm-HAN-488 | In situ unlabeled | In situ labeled 1.4-nm-594 | |||
Data collection | ||||||
Magnification | ×81,000 | ×81,000 | ×81,000 | ×81,000 | ×81,000 | ×105,000 |
Voltage (kV) | 300 | 300 | 300 | 300 | 300 | 300 |
Defocus range (μm) | −3 to −5 | −3 to −5 | −3 to −5 | −3 to −5 | −3 to −5 | −3 to −5 |
Pixel size (Å) | 1.068 | 1.068 | 1.068 | 1.068 | 1.068 | 0.876 |
High Dose 0° | 20 | 0 | 20 | 0 | 20 | 0 |
Total Electron exposure (e–/Å2) | 180 | 170 | 180 | 160 | 180 | 170 |
Tilt-range/step (°) | ±36 /2 | ±48/2 | ±36/2 | ±48/2 | ±36/2 | ±40/2 |
Tilt-scheme | Dose-symmetric, grouping 2 | |||||
Processing | ||||||
Symmetry imposed | C1 | C1 | C1 | C1 | ||
Initial particle images (no.) | 13,748 | 77,336 | 17,662 | 6,603 | ||
Final particle images (no.) | 1,743 | 10,759 | 2,7689 | 4,955 | ||
Map resolution (Å) | 15 | 11 | 11.2 | 18 Å | ||
Fourier shell correlation threshold | 0.143 | |||||
