Automated fiducial-based alignment of cryo-electron tomography tilt series in Dynamo

Structure. 2024 Oct 3;32(10):1808-1819.e4. doi: 10.1016/j.str.2024.07.003. Epub 2024 Jul 29.

Abstract

With the advent of modern technologies for cryo-electron tomography (cryo-ET), high-quality tilt series are more rapidly acquired than processed and analyzed. Thus, a robust and fast-automated alignment for batch processing in cryo-ET is needed. While different software packages have made available several approaches for automated marker-based alignment of tilt series, manual user intervention remains necessary for many datasets, thus preventing high-throughput tomography. We have developed a MATLAB-based framework integrated into the Dynamo software package for automatic detection of fiducial markers that generates a robust alignment model with minimal input parameters. This approach allows high-throughput, unsupervised volume reconstruction. This new module extends Dynamo with a large repertory of tools for tomographic alignment and reconstruction, as well as specific visualization browsers to rapidly assess the biological relevance of the dataset. Our approach has been successfully tested on a broad range of datasets that include diverse biological samples and cryo-ET modalities.

Keywords: contrast transfer function; cryo electron tomography; fiducial tracking; subtomogram averaging; tilt series alignment; tomogram reconstruction.

MeSH terms

  • Algorithms
  • Cryoelectron Microscopy* / instrumentation
  • Cryoelectron Microscopy* / methods
  • Electron Microscope Tomography* / instrumentation
  • Electron Microscope Tomography* / methods
  • Fiducial Markers
  • Humans
  • Image Processing, Computer-Assisted* / methods
  • Software*