The macroscale at which we measure, model and predict climate change does not align with the microscale at which small ectotherms experience climate. To understand climate's influence on biodiversity and potential ecological effects of climate change, more work is needed to understand how ectotherm physiology relates to microclimatic temperatures. Tree canopies are an example of a habitat that produces extreme microclimates, and arthropods in tropical forest canopies are threatened by extreme heat and warming. The situation in temperate canopies, however, is less clear. Conventional wisdom suggests that winter cold limits arboreal arthropod diversity in temperate forests, but because the canopy is less buffered from extreme temperatures, summer heat could also play a role. Heat- and cold-limited communities will respond differently to climate change, so this distinction is critical. Using the frameworks of the thermal adaptation hypothesis and thermal niche asymmetry, we asked whether arboreal ants were physiologically adapted to their extreme environment and whether summer heat or winter cold was more stressful. We tracked internal microclimates of ant nests in the canopy and on the ground over the seasonal cycle in temperate forests in North Carolina, USA. Then, we measured the heat (CT) and cold tolerance (CT) of worker ants in summer and spring and compared them with ants' experienced microclimates. Nests in the temperate canopy experienced hotter and colder extremes and more closely tracked air temperatures than ant nests on the ground. Arboreal ants partially adhered to the thermal adaptation hypothesis. They were more heat tolerant than ground-nesting species, but despite experiencing lower temperatures, they were less cold-tolerant. Ants acclimated their cold tolerance in line with seasonal changes, but heat tolerance was more phylogenetically constrained. Summer heat did not approach ants' heat tolerance in either stratum, but winter and spring lows in the canopy exceeded the cold tolerance of ants nesting there. By comparing microclimatic temperatures and thermal physiology, we suggest that winter cold-and not summer heat-likely limits arthropod diversity in the temperate canopy.