The visual system is known to be vital for cognition and perception in the feline and canine and much behavioral research for these species has used visual stimuli and focused on visual perception. There has been extensive investigations into the visual pathway in cats and dogs via histological and neurobiological methods, however to date, only one study has mapped the canine optic pathway in vivo.Advanced imaging methods such as diffusion MRI (DTI) have been routinely used in human research to study the visual system in vivo. This study applied DTI imaging methods to assess and characterize the optic pathway of feline and canine subjects in vivo. The optic nerve (ON), optic tract (OT), and optic radiation (OR) were successfully delineated for each species and the average volume and FA for each tract is reported. The application of DTI to map the optic pathway for canine and feline subjects provides a healthy baseline for comparison in future studies.