Mosquitofish (Gambusia affinis) is an example of a freshwater fish species whose remarkable diffusion outside its native range has led to it being placed on the list of the world’s hundred worst invasive alien species (International Union for Conservation of Nature). Here, we investigate mosquitofish shoaling tendency using a dichotomous choice test in which computer-animated images of their conspecifics are altered in color, aspect ratio, and swimming level in the water column. Pairs of virtual stimuli are systematically presented to focal subjects to evaluate their attractiveness and the effect on fish behavior. Mosquitofish respond differentially to some of these stimuli showing preference for conspecifics with enhanced yellow pigmentation while exhibiting highly varying locomotory patterns. Our results suggest that computer-animated images can be used to understand the factors that regulate the social dynamics of shoals of Gambusia affinis. Such knowledge may inform the design of control plans and open new avenues in conservation and protection of endangered animal species.