We conducted a national-scale assessment of mercury (Hg) bioaccumulation in aquatic ecosystems using dragonfly larvae as biosentinels, by developing a citizen science network to facilitate biological sampling. Implementing a carefully designed sampling methodology for citizen scientists, we developed an effective framework for landscape-level inquiry that might otherwise be resource limited. We assessed variation in dragonfly Hg concentrations across >450 sites spanning 100 US National Park Service units, and examined intrinsic and extrinsic factors associated with variation in Hg concentrations. Mercury concentrations ranged between 10.4-1,411 ng/g dry weight across sites and varied among habitat types. Dragonfly total Hg (THg) concentrations were up to 1.8-fold higher in lotic habitats than in lentic habitats, and 37% higher in waterbodies with abundant wetlands along their margins than those without wetlands. Mercury concentrations in dragonflies differed among families, but were correlated (R2>0.80) with each other, enabling adjustment to a consistent family to facilitate spatial comparisons among sampling units. Dragonfly THg concentrations were positively correlated with THg in both fish and amphibians from the same locations, indicating that dragonfly larvae are effective indicators of Hg bioavailability in aquatic food webs. Collectively, this continental-scale study demonstrates the utility of dragonfly larvae for estimating potential mercury risk to fish and wildlife in aquatic ecosystems and provides a framework for engaging citizen science as a component of landscape Hg monitoring programs.