Many early studies of prejudice adopted psychophysiological measures as a way to circumvent the limitations of self-report instruments. Despite serious methodological weaknesses, that literature consistently points to the value of physiological probes as nonreactive indexes of affective responses to target stimuli. Possible reasons for the virtual abandonment of psychophysiological approaches in the study of prejudice over the last 15 years are outlined, and their reintroduction is advocated on methodological and conceptual grounds. Theoretical perspectives and empirical research in a closely related area, the psychophysiology of emotion, are reviewed and the implications of this literature for the study of prejudice are discussed. Several psychophysiological approaches have been found valuable for assessing the valence and intensity of emotional responses. The availability of these tools, together with the shifting theoretical zeitgeist, make prejudice research ready for a return to psychophysiological methodologies. A multi-method prejudice assessment model is proposed and its theoretical and heuristic advantages are discussed.