The emerald ash borer, Agrilus planipennis Fairmaire, is an exotic invasive insect causing extensive mortality to ash trees, Fraxinus spp., in Canada and the United States. Detection of incipient populations of this pest is difficult because of its cryptic life stages and a multiyear time lag between initial attack and the appearance of signs or symptoms of infestation. We sampled branches from open-grown urban ash trees to develop a sample unit suitable for detecting low density A. planipennis infestation before any signs or symptoms are evident. The sample unit that maximized detection rates consisted of one 50-cm-long piece from the base of a branch ≥6 cm diameter in the midcrown. The optimal sample size was two such branches per tree. This sampling method detected ≈75% of asymptomatic trees known to be infested by using more intensive sampling and ≈3 times more trees than sampling one-fourth of the circumference of the trunk at breast height. The method is less conspicuous and esthetically damaging to a tree than the removal of bark from the main stem or the use of trap trees, and could be incorporated into routine sanitation or maintenance of city-owned trees to identify and delineate infested areas. This research indicates that branch sampling greatly reduces false negatives associated with visual surveys and window sampling at breast height. Detection of A. planipennis-infested asymptomatic trees through branch sampling in urban centers would provide landowners and urban foresters with more time to develop and implement management tactics.