We report the binding properties and subnuclear localization of [3H]hemicholinium-3 binding sites in human amygdala using quantitative autoradiography. Specific binding was saturable and high affinity (apparent Kd 2-11 nM). Binding was highest in the basolateral nucleus which receives dense cholinergic innervation from the basal forebrain. Binding closely approximated acetylcholinesterase reactivity. These data support [3H]hemicholinium-3 as a quantitative marker for cholinergic terminals in human brain.