Related Experiment Video
Updated: Jun 12, 2026

Single-Particle Cryo-EM Data Collection with Stage Tilt using Leginon
Published on: July 1, 2022
Fast and accurate marker-based projective registration method for uncalibrated transmission electron microscope tilt
Ho Lee1, Jeongjin Lee, Yeong Gil Shin
1Department of Radiation Oncology, Stanford University, Stanford, CA 94305-5847, USA. leeho@stanford.edu
Abstract:
This paper presents a fast and accurate marker-based automatic registration technique for aligning uncalibrated projections taken from a transmission electron microscope (TEM) with different tilt angles and orientations. Most of the existing TEM image alignment methods estimate the similarity between images using the projection model with least-squares metric and guess alignment parameters by computationally expensive nonlinear optimization schemes. Approaches based on the least-squares metric which is sensitive to outliers may cause misalignment since automatic tracking methods, though reliable, can produce a few incorrect trajectories due to a large number of marker points. To decrease the influence of outliers, we propose a robust similarity measure using the projection model with a Gaussian weighting function. This function is very effective in suppressing outliers that are far from correct trajectories and thus provides a more robust metric. In addition, we suggest a fast search strategy based on the non-gradient Powell's multidimensional optimization scheme to speed up optimization as only meaningful parameters are considered during iterative projection model estimation. Experimental results show that our method brings more accurate alignment with less computational cost compared to conventional automatic alignment methods.

