A workflow to combine cryo-EM data collected at different magnification.

The Scipion framework allows very flexible image-processing workflows to be generated and employed at electron-microscopy facilities, such that image acquisition can be monitored and possible problems detected, thereby enabling early decisions to be made on the fly. The streaming workflow can be very simple or extended, permitting the data resolution and heterogeneity to be estimated and adapted to the desire of the user and the microscope operator.

Installation of the EIGER X 1M detector onto an electron microscope and system calibration for data collection turns a transmission electron microscope into an electron diffraction instrument. Data can be collected and processed with a throughput that meets the requirements of a modern X-ray facility. The setup described here offers access to single-crystal structure determination from microcrystalline powder when large single crystals cannot be produced.

The study of coat protein assemblies on membranes by cryo-electron tomography and subtomogram averaging is discussed, outlining the methodological advancements that enabled the gain of crucial structural insight.