Dengue is currently the most important arboviral disease globally and is usually associated with built environments in tropical areas. Remotely sensed information can facilitate the study of urban mosquito-borne diseases by providing multiple temporal and spatial resolutions appropriate to investigate urban structure and ecological characteristics associated with infectious disease. In this study, coarse, medium and fine resolution satellite imagery (Moderate Resolution Imaging Spectrometer, Advanced Spaceborne Thermal Emission and Reflection Radiometer and QuickBird respectively) and ground-based data were analyzed for the Greater Puntarenas area, Costa Rica for the years 2002–04. The results showed that the mean normalized difference vegetation index (NDVI) was generally higher in the localities with lower incidence of dengue fever during 2002, although the correlation was statistically significant only in the dry season (r =−0.40; p = 0.03). Dengue incidence was inversely correlated to built area and directly correlated with tree cover (r = 0.75, p = 0.01). Overall, the significant correlations between dengue incidence and urban structural variables (tree cover and building density) suggest that properties of urban structure may be associated with dengue incidence in tropical urban settings.