The organization of ON and OFF responses and ocular dominance in the striate cortex of the tree shrew was electrophysiologically investigated by using flashed, stationary visual stimuli presented monocularly to either the ipsilateral or contralateral eye. We measured cortical multi‐unit activity at 25‐μm intervals with glass‐insulated platinum‐plated tungsten microelectrodes. Penetrations were made perpendicular to the cortical layers and the responses were quantitatively analyzed in layers IIIc to V. In sublayers IIIb, IIIc, and upper V, phasic responses of approximately equal magnitude occurred to both light ON and light OFF (ON‐OFF regions). In layer IV, tonic as well as phasic responses were often evoked by the flashed spot of light. In sublayer IVa stronger responses occurred to light ON than to light OFF (ON region) while in sublayer IVb stronger responses occurred to light OFF than to light ON (OFF region). In an ON region, the increased neural activity that occurred at light ON was often accompanied by a decrease in activity below baseline level at light OFF. A similar decrease often occurred in an OFF region at light ON. Recordings from the region of the cell‐sparse cleft in layer IV were characterized by ON‐OFF responses, signalling a transition zone between sublayers IVa and IVb. In addition, the responses to stimulation of the ipsilateral eye typically were very weak in the cleft region. In the other regions examined, the multi‐unit activity generally was driven binocularly with slightly greater responses being elicited by the contralateral eye. We conclude that the ON‐center and OFF‐center afferent pathways that are organized at the retinal level remain generally segregated in the tree shrew through the first synapse in the striate cortex. In addition, our recordings confirm that a horizontal organization of ocular dominance occurs in layer IV of the striate cortex in tree shrews. Copyright © 1986 Alan R. Liss, Inc.