A complete data processing workflow for CryoET and subtomogram averaging

Electron cryotomography (CryoET) is currently the only method capable of visualizing cells in 3D at nanometer resolutions. While modern instruments produce massive amounts of tomography data containing extremely rich structural information, the data processing is very labor intensive and results are often limited by the skills of the personnel rather than the data. We present an integrated workflow that covers the entire tomography data processing pipeline, from automated tilt series alignment to subnanometer resolution subtomogram averaging. Resolution enhancement is made possible through the use of per-particle per-tilt CTF correction and alignment. The workflow greatly reduces human effort and increases throughput and is capable of determining protein structures at state-of-the-art resolutions for both purified macromolecules and cells.