A method is presented to determine the spatial distribution of electrons in the focused beam of a scanning electron microscope (SEM). Knowledge of the electron distribution is valuable for characterizing and monitoring SEM performance, as well as for modeling and simulation in computational scanning electron microscopy. Specifically, it can be used to characterize astigmatism as well as study the relationship between beam energy, beam current, working distance, and beam shape and size. In addition, knowledge of the distribution of electrons in the beam can be utilized with deconvolution methods to improve the resolution and quality of backscattered, secondary, and transmitted electron images obtained with thermionic, FEG, or Schottky source instruments. The proposed method represents an improvement over previous methods for determining the spatial distribution of electrons in an SEM beam. Several practical applications are presented.