Membrane protein structural biology has made tremendous advances over the last decade but there are still many challenges associated with crystallization, data collection and structure determination. Two independent groups, Axford et al. [(2015), Acta Cryst. D71, 1228–1237] and Huang et al. [(2015), Acta Cryst. D71, 1238–1256], have published methods that make a major contribution to addressing these challenges.

An automated data-collection system named ZOO has been developed. This system enabled faster data collection, facilitated advanced data-collection and data-processing techniques, and permitted the collection of higher quality data.

The design and assembly of two 3D-printed holders for high-throughput in meso in situ fixed-target crystallographic data collection are described.

Recent developments in protein microcrystallography using synchrotron radiation are reviewed, including the use of a high-flux microbeam, consideration of radiation damage, sample-handling techniques, and data-collection strategies and their automation.

A method for performing high-throughput in situ serial X-ray crystallography with soluble and membrane proteins in the lipid cubic phase at cryogenic temperatures (100 K) is described. It works with nanogram to single-digit microgram quantities of protein and lipid (and ligand when present), and is compatible with both high-resolution native data collection and experimental phasing without the need for crystal harvesting.

Several oils were examined for use in the cleaning and cryoprotection of crystals in the lipidic cubic phase in terms of their effect on the crystal stability, the background scattering and the facilitation of the experiment.

A method for performing high-throughput in situ serial X-ray crystallography with soluble and membrane proteins in the lipid cubic phase is described. It works with microgram quantities of protein and lipid (and ligand when present) and is compatible with the most demanding sulfur SAD phasing.

A simple robust manual protocol for producing crystals in the lipidic cubic phase in less than an hour is described. It is designed to provide newcomers to the in meso method for crystallizing membrane proteins with experience of preparing, handling and growing crystals in the sticky and viscous lipidic mesophase.

It is shown that even a low dose (0.06 MGy) of synchrotron radiation absorbed by a protein crystal induces structural alterations. This finding raises a significant issue as it may question previously solved structures as well as highlighting and defining new limitations of protein X-ray crystallography.

A distorted structure originally described as a mono­clinic polymorph of meso-(E,E)-1,1′-[1,2-bis­(4-chloro­phen­yl)ethane-1,2-di­yl]bis­(phenyl­diazene) is better modeled as a threefold superposition of undistorted S,S and R,R enanti­omers with a smaller fraction of the meso isomer. All reasoning behind the reassessment is explained in detail.