In order to investigate gender differences in pain perception, the present study employed both a psychophysical and a psychophysiological measure. In experiment 1, 20 subjects rated the painfulness of 4 different levels of tonic pressure applied to their fingers using a verbally anchored categorization procedure. In general agreement with studies of pain threshold and tolerance, female subjects reported greater pain at high levels of stimulation, with no gender difference being evident at low pressure levels. In experiment 2, 16 different subjects were exposed to the same painful pressure stimuli while measuring their pupil reactions using infrared video pupillometry. The pupil dilations seen during the last 10 sec of the 20-sec pressure application turned out to be a highly significant indicator of pain intensity. When female and male subjects were compared on this measure, a similar divergent pattern as in the psychophysical data emerged, with female subjects showing greater pupil dilations at high pressure levels only. The fact that gender differences in pain perception can be demonstrated using an autonomic indicator of pain that is beyond voluntary control suggests that these differences reflect low-level sensory and/or affective components of pain rather than attitudinal or response-bias factors.