Hemlock woolly adelgid, Adelges tsugae Annand (Hemiptera: Adelgidae), is an exotic pest of eastern hemlock, Tsuga canadensis (L.) Carrière (Pinales: Pinaceae), in the eastern United States. Two commonly used insecticides to manage adelgid are imidacloprid, a systemic neonicotinoid insecticide, and horticultural oil, a refined petroleum oil foliar spray. We have investigated the influence of imidacloprid and horticultural oil on spider abundance at different canopy strata in eastern hemlock. In total, 2,084 spiders representing 11 families were collected from the canopies of eastern hemlock. In beat-sheet and direct observation samples, the families Theridiidae, Araneidae, Salticidae, and Anyphaenidae were the most abundant. Significantly higher numbers of spiders were recorded on untreated control trees compared with trees treated with imidacloprid using soil drench and soil injection applications. Spider abundance in trees injected with imidacloprid and horticultural oil applications did not significantly differ from control trees. Spider abundance was significantly greater in the top and middle strata of the canopy than in the bottom stratum, where imidacloprid concentrations were the highest. Regression analysis showed that spider abundance was inversely associated with imidacloprid concentration. This research demonstrates that imidacloprid, when applied with selected methods, has the potential to result in reductions of spider densities at different strata. However, slight reductions in spider abundance may be an acceptable short-term ecological impact compared with the loss of an untreated hemlock and all the associated ecological benefits that it provides. Future studies should include investigations of long-term impact of imidacloprid on spiders associated with eastern hemlock.