Aphaenogaster mariae Forel is a rarely encountered North American arboreal ant that has eluded collectors for decades. Here, we provide the first formal documentation of a whole colony collection of the species found seventeen meters high in the canopy of the North Carolina Piedmont. We discovered a mature colony with more than 1000 individuals, including workers, alate reproductives, immatures, and intercastes. We present the first images of the males, larvae, pupae, and intercaste workers, redescribe the male, and provide natural history insights and colony demographics for this elusive species. Our collections suggest that A. mariae occurs at low densities consistent with its putative socially parasitic life history. Although much remains to be learned about this species, our results expand knowledge of its life history and facilitate future nest discovery and identification.