Ganglion cells in the albino rat retina were retrogradely labeled with the fluorescent dye, diamidino-yellow, from the superior colliculus. Preembedding and postembedding immunocytochemical techniques were employed in conjunction with computer-assisted image processing to visualize SP- and GABA-immunoreactivity. Examination of flatmount and sectioned retinas revealed that approximately 3% of the ganglion cells projecting to the contralateral superior colliculus exhibit SP-immunoreactivity. Moreover, these cells were found to comprise a subpopulation of the GABA-immunoreactive cells projecting to the rat tectum.