Automatic detection of alignment errors in cryo-electron tomography

Cryo-electron tomography is an imaging technique that allows the study of the three-dimensional structure of a wide range of biological samples, from entire cellular environments to purified specimens. This technique collects a series of images from different views of the specimen by tilting the sample stage in the microscope. Subsequently, this information is combined into a three-dimensional reconstruction. To obtain reliable representations of the specimen of study, it is mandatory to define the acquisition geometry accurately. This is achieved by aligning all tilt images to a standard reference scheme. Errors in this step introduce artifacts into the final reconstructed tomograms, leading to loss of resolution and making them unsuitable for detailed sample analysis. This publication presents algorithms for automatically assessing the alignment quality of the tilt series and their classification based on the residual errors provided by the alignment algorithms. If no alignment information is available, a set of algorithms for calculating the residual vectors focused on fiducial markers is also presented. This software is accessible as part of the Xmipp software package and the Scipion framework. [Display omitted] •Misaligned tomographic reconstructions are unsuitable for the study of the sample.•Efficient and robust automatic tilt-series alignment assessment.•Avoid the tedious and error-prone task of manually inspecting tilt-series alignment.•Adaptative configuration based on the strictness of the scientist.•Residual calculation algorithm provided if they are missing from the alignment.