Figure 2. Final refined structure of lysozyme at 2.5 Å from Continuous Rotation MicroED data.

(a-d) A representative region of the final refined structure of lysozyme originating from the 2-crystal data set is shown, with the 2F_obs-F_calc density map (a; contoured at 1.0σ) showing well defined density around the backbone and sidechains. The final 3D structure is also shown in Supplementary Video 3. To test any potential model bias, residues 27 through 36 were removed from the final refined model and the incomplete model was used to phase and refine the original data. The 2F_obs-F_calc map (contoured at 1.0σ) without the deleted residues (b) shows clearly defined density for both backbone and sidechains where the missing residues (shown in yellow) could easily be placed (c). The F_obs-F_calc map (contoured at 3σ) also shows very strong density for the deleted residues (d). The strong density for the missing residues in the 2F_obs-F_calc and F_obs-F_calc maps indicate the final map does not suffer from model bias. (e-f) Final refined structure of lysozyme at 2.5 Å resolution using data originating from a single crystal. The 2F_obs-F_calc density map (e; contoured at 1.0σ) shown around residues 20-35 shows well-defined density around both backbone and sidechains. The F_obs-F_calc (f; contoured at ±3σ) shows no clear differences between the observed data and the model.