The adult human neocortex contains a dense net of axons and perikarya which yield an acetylcholinesterase-rich enzymatic reaction pattern in histochemical experiments. We employed a monoclonal antibody to human acetylcholinesterase and a method for the concurrent visualization of histochemical and immunohistochemical reaction-products to explore the relationship between immunological and enzymatic markers of acetylcholinesterase. We observed that the cortical axons and perikarya with a histochemically determined acetylcholinesterase-rich enzymatic activity also contain acetylcholinesterase-like immunoreactivity. This was especially informative for the intracortical acetylcholinesterase-rich perikarya of layers III and V since these neurons require prolonged incubations for histochemical detection and since they are not conspicuous in other animal species. The availability of a reliable immunohistochemical method makes it possible to investigate the distribution of the acetylcholinesterase enzyme molecule independent of its enzymatic activity.