Objective : There has been inconsistency about definition of the temporal stem despite of several descriptions demonstrating its microanatomy using fiber dissection and/or diffusion tensor tractography. This study was designed to clarify three dimensional configurations of the temporal stem. Methods : The fronto-temporal regions of several formalin-fixed human cerebral hemispheres were dissected under an operating microscope using the fiber dissection technique. The consecutive coronal cuts of the dissected specimens were made to define the relationships of white matter tracts comprising the temporal stem and the subcortical gray matters (thalamus, caudate nucleus, amygdala) with inferior limiting (circular) sulcus of insula. Results : The inferior limiting sulcus of insula, limen insulae, medial sylvian groove, and caudate nucleus/amygdala were more appropriate anatomical structures than the roof/dorso-lateral wall of the temporal horn and lateral geniculate body which were used to describe previously for delineating the temporal stem. The particular space located inside the line connecting the inferior limiting sulcus of insula, limen insulae, medial sylvian groove/amygdala, and tail of caudate nucleus could be documented. This space included the extreme capsule, uncinate fasciculus, inferior occipito-frontal fasciculus, anterior commissure, ansa peduncularis, and inferior thalamic peduncle including optic radiations, whereas the stria terminalis, cingulum, fimbria, and inferior longitudinal fiber of the temporal lobe were not passing through this space. Also, this continued posteriorly along the caudate nucleus and limiting sulcus of the insula. Conclusion : The temporal stem is white matter fibers passing through a particular space of the temporal lobe located inside the line connecting the inferior limiting sulcus of insula, limen insulae, medial sylvian groove/amygdala, and tail of caudate nucleus. The three dimensional configurations of the temporal stem are expected to give the very useful anatomical and surgical insights in the temporal lobe.