Across their range jaguars Panthera onca are important conservation icons for several reasons: their important role in ecosystems as top carnivores, their cultural and economic value, and their potential conflicts with livestock. However, jaguars have historically been difficult to monitor. This paper outlines the first application of a systematic camera trapping methodology for abundance estimation of jaguars. The methodology was initially developed to estimate tiger abundance in India. We used a grid of camera traps deployed for 2 months, identified individual animals from their pelage patterns, and estimated population abundance using capture-recapture statistical models. We applied this methodology in a total of five study sites in the Mayan rainforest of Belize, the Chaco dry forest of Bolivia, and the Amazonian rainforest of Bolivia. Densities were 2.4–8.8 adult individuals per 100 km2, based on 7–11 observed animals, 16–37 combined ‘captures’ and ‘recaptures’, 486–2,280 trap nights, and sample areas of 107–458 km2. The sampling technique will be used to continue long-term monitoring of jaguar populations at the same sites, to compare with further sites, and to develop population models. This method is currently the only systematic population survey technique for jaguars, and has the potential to be applied to other species with individually recognizable markings.