Validation of 3D cryoEM single particle reconstruction correctness and handedness with Ewald's sphere correction.

The correct description of quantum scattering places the observed scattering contributions on the Ewald's sphere and its Friedel mate. In electron microscopy, due to the large radius of the Ewald's sphere, these contributions are typically merged during data analysis. We present an approach that separates and factorizes these contributions into inversion-symmetric and inversion-antisymmetric components. The correlations between reconstructions derived from these symmetric and antisymmetric components enable the automatic determination of handedness and provide additional validation for the quality of 3D reconstructions. These correlations are robust enough to be routinely used in single-particle reconstructions, even at resolutions below the limit where the curvature of the Ewald's sphere affects the overall signal-to-noise ratio.