BEAM IMAGE-SHIFT ACCELERATED DATA ACQUISITION FOR SINGLE-PARTICLE ELECTRON CRYO-MICROSCOPY WITH SPHERICAL ABERRATION CORRECTION

Abstract

The throughput of electron cryo-microscopy (cryo-EM) can be improved by collecting data using beam-image shift instead of the traditional stage-shift method. Despite successful implementation of beam-image shift on various cryo-EM instruments, it remains unknown how beam-image shift affects the quality of data collected on spherical aberration (Cs)-corrected cryo-EM instruments. To investigate this, a cryo-EM dataset for Thermoplasma acidophilum 20S (T20S) proteasome was collected on a 300 kV Cs-corrected transmission electron microscope (TEM) using beam-image shift. Analyzing the dataset in two different image-processing software, RELION and CryoSPARC demonstrated that there are minimal differences between the resolutions of T20S structures generated from datasets collected using with and without beam-image shift. In addition, findings of this study demonstrated that data collected using with and without beam-image shift can be combined and processed together to correct for image abberations.