We have succeeded in determining the charge of individual colloidal particles with resolution higher than the elementary charge. The number of elementary charges on a particle is obtained from the analysis of optical tracking data of weakly charged silica spheres in an electric field in a nonpolar medium. The analysis also yields an accurate value of the particle size. Measurement of the charge as a function of time reveals events in which the particle loses or gains an elementary charge due to ionization or recombination processes at the surface.