Abstract Background Given increasing investment in new cycling infrastructure, it is important to understand its impacts. The Sydney Transport and Health Study evaluates a new 2.4 km bi-directional separated bicycle path in inner-Sydney. This paper describes the users of the new bicycle path, and examines its short-term impacts upon cycling behaviour and perceptions of the local environment. Methods Data were collected from two bike counts at two intersections on the new bicycle path in the intervention area in 2013 and 2014. On-line surveys collected individual participant data in the intervention area and a similar comparison area before the bicycle path was built (2013), and 12 months later (four months after completion) ( n  = 512). The data included self-reported cycling behaviour, use of the new bicycle path and perceptions of changes in the local environment. Results Bike counts at two sites on the new bicycle path reported an increase of 23 % and 97 % respectively at 12 months. However, among the participants in the cohort, there was no change in the self-reported weekly frequency of cycling. One in six (approximately 15 %) participants reported using the new bicycle path, with most users (76 %) living in the intervention area. Bicycle path users were most likely to be frequent riders (at least weekly) [adjusted odds ratio (AOR) = 7.50, 95 % CI 3.93–14.31], be a high intensity recreational rider (AOR = 4.38, 95 % CI 1.53–12.54) or a low intensity transport rider (AOR = 2.42, 95 % CI 1.17–5.04) and live closer to the bicycle path (AOR = 1.24, 1.13–1.37). Perceptions that the neighbourhood was more pleasant, that there were more people walking and cycling were significantly higher in the intervention area at 12 months (both P values