Purpose The aim of this study was to describe normal ocular morphology and establish parameters for select diagnostics in a population of Panamanian Golden Frogs (PGF) Atelopus zeteki maintained at the Detroit Zoo. Method A total of 22 toads free of ocular disease were included in this study. Ophthalmic diagnostic tests included as follows: endodontic absorbent paper point tear test (EAPPTT), assessment of intraocular pressures (IOP), Rose Bengal stain, palpebral fissure width, blink rate and corneal cultures. Histology was performed on stored postmortem samples. Results Panamanian Golden Frogs have a distinct brow ridge and mobile third eyelid. Biomicroscopy revealed avascular, convex corneas with a shallow anterior chamber. Iris color ranged from yellow to green with dark pigmentation of the peripheral iris. Pupil shape was round. Posterior segment and fundus could not be visualized. Total number of blinks observed over a 10‐minute period was one. Mean EAPPTT was 2 ± 1 mm and 3 ± 1 mm in males and females, respectively (P). Mean IOP using rebound tonometry in horizontal and vertical restraint positions were 21.6 ± 2.28 mm Hg and 17.45 ± 3.66 mm Hg, 21.85 ± 2.7 mm Hg and 17.5 ± 3.93 mm Hg in males and females, respectively. Mean palpebral fissure width was 5.02 ± 0.46 mm and 6.12 ± 0.39 mm in males and females, respectively (P). Enterococcus faecalis was isolated from one clinically normal toad. Conclusions Tear production and blink rate were low and not significantly different between males and females. Intraocular pressure was significantly greater in male toads (P value <0.01). Females had a significantly larger palpebral fissure width when compared to males (P value <0.01) 5819. The ophthalmic parameters established in this study can aid in the evaluation and diagnosis of eye diseases in PGF.