Abstract A method for evoking physiological responses by microinjection of sodium glutamate solution into localized regions of the central nervous system (CNS) is described. The major advantage of this method is that the cell bodies or dendritic processes of neurones within the injection site are excited, whereas axons of passage are unaffected. It was demonstrated that injections of minute volumes (50–100 nl) of 0.5 M glutamate solution into selected sites within the medulla or midbrain of anaesthetized or conscious animals, respectively, elicited marked autonomic, somatomotor or behavioural responses, depending on the injection site. In contrast, glutamate microinjection into fibre tracts failed to elicit any response, whereas electrical stimulation applied at the same sites elicited marked responses. The degree of localization of the glutamate stimulus and the relation between glutamate concentration and magnitude of evoked response are described. It is concluded that this method is a very effective means of selectively stimulating cell bodies within highly localized regions of the CNS. Further, by using this method in combination with focal electrical stimulation, it is possible in some cases to provide evidence that a response arises from excitation of axons of passage rather than cell bodies.