Amyloid structure determination in RELION is increasingly popular, but may suffer from refinements becoming stuck in local minima. This paper describes a new method for initial model generation that may alleviate this problem.

This article presents an overview of the development of two basic software libraries for image manipulation and data visualization in cryo-EM: emcore and emvis.

Two approaches for the selection of filaments from cryo-EM micrographs are described.

A new module of the ARP/wARP suite for automated model building into cryo-EM maps is presented.

The model-building software Buccaneer has been repurposed based on existing methods for building models into cryo-EM maps. These approaches are implemented in the latest CCP-EM model-building pipeline (version 1.4.0).

A method is presented for high-speed low-volume cryo-EM specimen preparation with a device constructed from readily available components.

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

Sample preparation within single-particle cryo-electron microscopy can still be a significant bottleneck, with issues in reproducibility, ice quality and sample loss. New approaches have recently been reported that use spraying or pin printing instead of the traditional blotting approach. Here, experience in the use of different nozzle designs and spraying regimes is reported together with their influence on the resulting spray and grid quality.

The concept of statistical signal detection by controlling the false-discovery rate (FDR) to aid the atomic model interpretation of cryo-EM density maps is reviewed. The recommended usage of the FDR software tool is presented together with its successful integration into the CCP-EM suite.