Fig. 1

Procedures and implications for obtaining a sub-2 Å resolution reconstruction of AAV2_L336C. a Fourier shell correlation (FSC) curves showing independent contributions of each operation to the final resolution. The resolution cut-off of 0.143 (gray dotted line) is indicated. Each FSC curve was generated by “turning off” one of the respective operations from the final refinement (1A). Correcting for rotational beam-induced movement (1B) improves the FSC at low spatial frequency. Discriminating particles based on their score (3) led to a lower resolution reconstruction (1.89 Å) as compared to equally weighting all particles (1.84 Å). Cumulative loss by “turning off” all the tested operations is indicated by (7). b ResLog⁶⁰ plot of the dataset. Five replicates were performed for each data point with random particle distributions and the standard deviation is plotted. Linear regression was fitted and the b-factor⁴⁵ indicated. c Fourier shell correlation (FSC) curves describing the half-map (blue solid line) and map-to-model (purple solid line) resolutions, as well as a histogram of directional resolutions sampled evenly over the 3DFSC⁵⁹ (yellow), and corresponding sphericity value. The resolution cut-offs of 0.143 (blue dotted line) and 0.5 (purple dotted line) were used respectively. d 1.86 Å reconstruction of the AAV2 viral capsid using single-particle cryo-EM, colored by local resolution⁵⁶. Slices through the reconstruction displayed in sub-panels start from the capsid center (left) move progressively towards the front (top right and bottom right). These show few deviations from the global resolution, with the best local resolution at 1.78 Å (primarily located inside the core region of the capsid shell). Both scales correspond to a distance of 100 Å