Cryo-electron microscopy (cryo-EM) has witnessed radical progress in the past decade, driven by developments in hardware and software. While current software packages include processing pipelines that simplify the image-processing workflow, they do not prioritize the in-depth analysis of crucial metadata, limiting troubleshooting for challenging data sets. The widely used RELION software package lacks a graphical native representation of the underlying metadata. Here, two web-based tools are introduced: relion_live.py, which offers real-time feedback on data collection, aiding swift decision-making during data acquisition, and relion_analyse.py, a graphical interface to represent RELION projects by plotting essential metadata including interactive data filtration and analysis. A useful script for estimating ice thickness and data quality during movie pre-processing is also presented. These tools empower researchers to analyse data efficiently and allow informed decisions during data collection and processing.

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S2059798323010902/ic5122sup1.mp4
Supplementary Video S1. relion_live.py, a web-based dashboard to follow on-the-fly pre-processing. The relion_live.py dashboard aggregates the pre-processing results of the relion_prep scheme. By clicking on each of the scattered points, the micrograph and its corresponding CTF estimation are displayed. The video illustrates how, by interacting with the plots, a data set can be explored using the display gallery. Applying manual thresholds allows micrographs that mostly contain carbon foil or bad-quality ice to be discarded.

Moving Picture Experts Group (MP4) video file https://doi.org/10.1107/S2059798323010902/ic5122sup2.mp4
Supplmentary Video S2. Volumes and classes display via relion_display or Chimera from the relion_analyse.py dashboard. When inspecting a 2D or 3D Classification or 3D Refinement job, the classes and volumes can be readily displayed using relion_display or the local installation of Chimera or ChimeraX.