The lateralization of emotion has received a great deal of attention over the last few decades, resulting in two main theories. The Right Hemisphere Theory states that the right hemisphere is primarily responsible for emotional processes, while the Valence Theory suggests that the right hemisphere regulates negative emotion and the left hemisphere regulates positive emotion. Despite the important implications of these theories for the evolution of emotion processes, few studies have attempted to assess the lateralization of emotion in non-human primates. This study uses the novel technique of measuring tympanic membrane temperature (Tty) to assess asymmetries in the perception of emotional stimuli in chimpanzees. The tympanic membrane is an indirect, but reliable, site from which to measure brain temperature, and is strongly influenced by autonomic and behavioral activity. Six chimpanzees were shown positive, neutral, and negative emotional videos depicting scenes of play, scenery, and severe aggression, respectively. During the negative emotion condition, right Tty was significantly higher than the baseline temperature. This effect was relatively stable, long lasting, and consistent across subjects. Temperatures did not change significantly from baseline in the neutral or positive emotion condition, although a significant number of measurements showed increased left Tty during the neutral emotion condition. These data suggest that viewing emotional stimuli results in asymmetrical changes in brain temperature, in particular increased right Tty during the negative emotion condition, evidence of emotional arousal in chimpanzees, and in providing partial support of both the Right Hemisphere and Valence Theories of emotional lateralization in our closest living ancestor.