ABSTRACT Introduction Current physiological measures of sexual arousal are intrusive, hard to compare between genders, and quantitatively problematic. Aim To investigate thermal imaging technology as a means of solving these problems. Methods Twenty-eight healthy men and 30 healthy women viewed a neutral film clip, after which they were randomly assigned to view one of three other video conditions: (i) neutral (N=19); (ii) humor (N=19); and (iii) sexually explicit (N=20). Main Outcome Measures Genital and thigh temperatures were continuously recorded using a TSA ImagIR camera. Subjective measures of sexual arousal, humor, and relaxation were assessed using Likert-style questions prior to showing the baseline video and following each film. Results Statistical (Tukey HSD) post-hoc comparisons (P <0.05) demonstrated that both men and women viewing the sexually arousing video had significantly greater genital temperature (mean=33.89°C, SD=1.00) than those in the humor (mean=32.09°C, SD=0.93) or neutral (mean=32.13°C, SD=1.24) conditions. Men and women in the erotic condition did not differ from each other in time to peak genital temperature (men mean=664.6 seconds, SD=164.99; women mean=743 seconds, SD=137.87). Furthermore, genital temperature was significantly and highly correlated with subjective ratings of sexual arousal (range r =0.51–0.68, P <0.001). There were no significant differences in thigh temperature between groups. Conclusion Thermal imaging is a promising technology for the assessment of physiological sexual arousal in both men and women.