An open-source platform for light-coupled cryo-electron microscopy specimen preparation is presented.

Methods for obtaining the maximum out of a cryo-EM data-collection session are described.

CM01 is a recently opened cryo-electron microscopy facility located at the European Synchrotron and is used by the ESRF's international community for structural biology.

A new method for accurate real-time defocus modulation in electron microscopy that will improve the configurability of the technique is presented.

An overview is given of the new CCP-EM software suite, including the underpinning framework, the current functionality and the distribution procedure. This first version of the suite has a focus on building and refining atomic models.

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.

Utilities for atomic model validation in the CCP-EM software suite are discussed. An assessment of SARS-CoV-2 structures reflects a strong bias in model refinement towards geometric restraints when compared with agreement with the data.

The 3.1 Å resolution single-particle cryo-electron microscopy structure of the RNA-bound Ebola virus nucleoprotein helical filament provides molecular details of protein–protein and protein–RNA interactions.

RSF1 changes the chromatin-remodeling mode of SNF2h. The RSF complex does not have a nucleosome-recentering capacity and a `critical distance' exists during the action of RSF in vitro; this distance is about 24 base pairs. The RSF complex loosens parts of the nucleosome DNA and undergoes conformational change on linker DNA stimulation.

Virtual reality-specific tools for model building are possible, and can provide an order-of-magnitude speedup over mouse-and-keyboard tools in certain situations.