An image processing pipeline for electron cryo‐tomography in RELION‐5

• Published in: FEBS open bio Vol. 14; no. 11; pp. 1788 - 1804
• Main Authors: Burt, Alister, Toader, Bogdan, Warshamanage, Rangana, Kügelgen, Andriko, Pyle, Euan, Zivanov, Jasenko, Kimanius, Dari, Bharat, Tanmay A. M., Scheres, Sjors H. W.
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01.11.2024; John Wiley and Sons Inc; Wiley
• Subjects: Automation; Bioinformatics; Cryoelectron Microscopy - methods; Electron Microscope Tomography - methods; Electron microscopes; Electron Microscopy; electron tomography; Image processing; Image Processing, Computer-Assisted - methods; Imaging, Three-Dimensional - methods; Metadata; Method; Microscopy; Models, Molecular; Open source software; Software; Structural Biology; Tomography; User interface; image processing; electron microscopy; software; electron tomography
• ISSN: 2211-5463; 2211-5463
• DOI: 10.1002/2211-5463.13873

Electron tomography of frozen, hydrated samples allows structure determination of macromolecular complexes that are embedded in complex environments. Provided that the target complexes may be localised in noisy, three‐dimensional tomographic reconstructions, averaging images of multiple instances of these molecules can lead to structures with sufficient resolution for de novo atomic modelling. Although many research groups have contributed image processing tools for these tasks, a lack of standardisation and interoperability represents a barrier for newcomers to the field. Here, we present an image processing pipeline for electron tomography data in RELION‐5, with functionality ranging from the import of unprocessed movies to the automated building of atomic models in the final maps. Our explicit definition of metadata items that describe the steps of our pipeline has been designed for interoperability with other software tools and provides a framework for further standardisation. We present an image processing pipeline for electron cryo‐tomography data in RELION‐5 with functionality ranging from unprocessed movie import to the automated building of atomic models in final maps. The metadata describing the pipeline steps is aimed at standardisation and interoperability with other software, while the performance improvements and visual tools lead to short processing times and ease of use.