Rapid tilt-series method for cryo-electron tomography: Characterizing stage behavior during FISE acquisition

We and others recently developed rapid tilt-series acquisition methods for cryo-electron tomography on a Titan Krios G3i equipped with a single axis holder and a K-series direct electron detector and showed that one of these, the fast-incremental single exposure (FISE) method, significantly accelerates tilt-series acquisition when compared to traditional methods while preserving the quality of the images. Here, we characterize the behavior of our single axis holder in detail during a FISE experiment to optimally balance data quality with speed. We explain our methodology in detail so others can characterize their own stages, and conclude with recommendations for projects with different resolution goals.

Automated summary

A rapid tilt-series acquisition method has been developed to significantly accelerate the acquisition speed while maintaining stable image quality. By detailing the behavior of the single axis holder, an optimal solution has been provided to balance data quality and speed.