Assessing Map Quality (cryo-EM)

Why

The first step after data processing is analysis to assess data quality and detect any pathologies that might thwart structure solution. The quality of the reconstruction, and therefore the interpretability of a cryo-EM map, can deteriorate from many causes including structural heterogeneity, radiation damage, and beam-induced sample movement.

Data quality is typically expressed by the resolution, but this term has a different meaning for cryo-EM than crystallographic experiments. For a cryo-EM map, the overall resolution (dFSC) is usually defined as the maximum spatial resolution at which the information content of the map is reliable.

The cryo-EM resolution is typically obtained by analyzing the Fourier shell correlation (FSC) for two cryo-EM half-maps binned in resolution shells. If the macromolecule is structurally heterogeneous, a single resolution value is adequate. 'Local resolutions' can be also assigned to different map regions.

How

The phenix.mtriage program estimates the resolution of cryo-EM maps using several approaches, and then provides a summary of the map statistics. The tool requires only a full cryo-EM map (in CCP4, MRC, or other related format). However, you can obtain additional information by inputting two half-maps and/or an atomic model (in PDB or mmCIF format).

How to use the phenix.mtriage: Click here

Common issues

Program stops: All input maps are expected to be on the same grid. Also, box information (e.g., unit cell of CRYST1 record in PDB files) must match across all of the inputs.

Related programs and Documentation