A new and comprehensive resource is described that contains validation information for all cryo-EM structures that are available in the public archives, EMDB and PDB, based on recommendations from the cryo-EM community.

In this article, it is shown that high-throughput strategies for tomographic data acquisition combined with unsupervised techniques for image analysis provide the foundation for closing the resolution gap between the high-resolution strategies used to study molecular assemblies reconstituted in vitro and techniques for in situ structure determination.

Time-resolved cryo-EM is an emerging technique in structural biology that allows the user to capture structural states which would otherwise be too transient for standard methods. There has been a resurgence in technical advancements in this field in the last five years and this review provides a summary of the technical highlights.

New computational biophysics pipelines for analysing the global dynamics of structural ensembles and large, dynamic complexes resolved by cryoEM are reviewed.

A new method, checkMySequence, for the fast and automated detection of register errors in protein models built into cryo-EM reconstructions is presented.

Single-particle analysis (SPA) by cryo-electron microscopy comprises the estimation of many parameters along its image-processing pipeline. Overfitting observed in SPA is normally due to misestimated parameters, and the only way to identify these is by comparing the estimates of multiple algorithms or, at least, multiple executions of the same algorithm.