Abstract Modern low-voltage scanning transmission electron microscopes (STEMs) have been invaluable for the atomic scale characterization of two-dimensional (2D) materials. Nevertheless, the observation of intrinsic structures of semiconducting and insulating 2D materials with 60 kV-microscopes has remained problematic due to electron radiation damage. In recent years, ultralow-voltage microscopes have been developed with the prospects of minimizing radiation damage of such 2D materials, however, to date only ultralow-voltage TEM investigations of semiconducting and insulating 2D materials have been reported, but similar results using STEM, despite being more widely adopted, are still missing. Here we report a quantitative analysis of radiation damage and beam-induced defect dynamics in semiconducting 2D WS₂ during 30 kV and 60 kV-STEM imaging, particularly by recording atomic resolution electrostatic potential movies using integrated differential phase contrast to visualize both the light sulfur and heavy tungsten atoms. Our results demonstrate that electron radiation damage of 2D WS₂ aggravates by a factor of two when halving the electron beam energy from 60 keV to 30 keV, from which we conclude electronic excitation and ionization to be the dominant mechanism inducing defects and damage during low-voltage STEM imaging of semiconducting 2D materials.