This study established ophthalmic reference values and characterized ocular lesions in two captive populations of boreal owls, including 46 eyes of 23 great grey owls (Strix nebulosa) and 38 eyes from 19 snowy owls (Bubo scandiacus). A complete ophthalmologic exam was conducted, including neuro-ophthalmic reflexes, Schirmer tear test I (STT-I), intraocular pressure (IOP) using rebound tonometry, fluorescein staining, horizontal corneal measurements using Jameson calipers, direct and indirect ophthalmoscopy, and ocular ultrasound biometry. Eyes with an STT of <5 mm/min, outliers, and eyes with severe diseases were excluded from reference value analysis. No statistically significant differences were found between right or left eyes in either species or among individuals in different age groups and sexes. Mean intraocular pressures and Schirmer tear tests were also not statistically significantly different between great grey owls and snowy owls (IOP: 9.6 ± 2.6 mm Hg and 9.1 ± 1.9 mm Hg, respectively, and STT-I: 9.8 ± 2.8 mm/min and 9.8 ± 2.4 mm/min, respectively). However, snowy owls overall had a significantly larger eye than did great grey owls, reflected in corneal diameters (23.4 ± 1 vs. 20.0 ± 0.8 mm, respectively) and sonographic biometry. In both species, the most common ocular lesions included keratitis, cataracts, chorioretinal lesions, and abnormal pecten. Establishment of reference ocular parameters will help wildlife veterinarians and rehabilitators determine an appropriate treatment plan and will aid in correctly identifying the presence of ocular disease.