Many cities have promoted decarbonized transportation modes to mitigate climate change, reduce air pollution and promote healthy behaviors. However, cyclists may be particularly exposed to higher concentrations of on-road air pollutants than other commuters due to their proximity to traffic, higher breathing rates, and prolonged commutes. In addition, there is scarce information analyzing the geographic exposure levels of cyclists in medium-sized urban agglomerations of Latin American cities. We aimed to assess cyclists' exposure to PM2.5 at the spatio-temporal level using low-cost sensors in the Mendoza Metropolitan Area, Argentina. We calculated PM2.5 inhalation doses (IDs) for different routes with distinctive characteristics, considering different age ranges and gender of cyclists. The dose was represented as temporally and spatially disaggregated exposure maps, one of the first to represent it in Latin America using this method. All link types analyzed exhibited significant differences in PM2.5 concentrations, although the most frequent concentrations were less than 5 µg m−3, with secondary peaks of 6.5 and 9 µg m−3. As expected, ID increases with age and is greater in males than in females. Our findings further reinforce the fact that route choice (busy vs. quiet) and time of day (peak vs. off-peak) appreciably affect the pollutant exposure of cyclists. All these results could be helpful in the selection of alternative cycling routes with lower PM2.5 at different hours of day but also enable us to investigate further implications of exposure to PM2.5 for the health of urban bicycle commuters.