Large carnivores are vulnerable to road effects because of their great mobility and extensive spatial requirements for survival. Wildlife crossing structures have mitigated harmful effects of roads for ungulate species, but there is limited information on how effective these structures are for large predators. We investigated the response of cougars Puma concolor to wildlife crossing structures along 45 kilometers of the Trans-Canada highway in Banff National Park, Alberta, Canada. Twenty-two crossing structures were monitored year-round for wildlife passage during 1996–2000. Cougar consistently used the wildlife crossing structures more than expected during winter months and less than expected during the summer. There was a significant positive correlation between passages made by cougar through wildlife crossing structures and those made by mule deer Odocoileus hemionus and white-tailed deer O. virginianus. There was no correlation between cougar and human use of the wildlife crossing structures. Cougar use of the five structure types differed from that expected. Open-span bridge underpasses were used more than expected, whereas creek bridge underpasses were used in proportion to their availability. All other crossing structure types were used significantly less than expected. The wildlife crossing structures that received the highest numbers of cougar passages were those situated close to high quality cougar habitat. The pattern of structure use was partly explained by the quality and distribution of cougar habitat near the structures as opposed to their physical features. Our results indicated that cougars tended to use underpasses more than wildlife overpass structures, and our study documents that cougars used crossing structures in a way that ensures habitat connectivity.