Ocular diseases reported in frogs include uveitis and glaucoma, which are associated with changes in intraocular pressure (IOP). The objectives of this study were to characterize the normal IOP for White's tree frogs (Litoria caerulea) using two types of rebound tonometers, and to assess whether time of day or method of restraint affected IOP. Eighteen conscious, unrestrained, ophthalmologically normal frogs were used to measure IOP using TonoVet® and TonoLab® tonometers, at three time points during the day. In a subset of 12 frogs, IOP was measured while under manual restraint using the TonoVet. Anesthesia was induced in 9 frogs using two different concentrations of MS-222 (0.5 g/L and 2 g/L) in order to evaluate for changes in IOP with the TonoVet. Mean (± SD) IOP values for the TonoLab (16.8 ± 3.9 mm Hg) were significantly higher than TonoVet values (14.7 ± 1.6 mm Hg; P < 0.01). TonoVet IOP values did not significantly change with time of day. TonoLab values were significantly lower in the evening (1600–1800; 14.5 ± 3.1 mm Hg), compared with morning and midday measurements (0800–1000 and 1200–1400; 18.0 ± 3.8 mm Hg; P < 0.01). Manually restrained frogs had significantly lower IOP (13.4 ± 1.5 mm Hg) compared with unrestrained frogs (15.3 ± 1.2 mm Hg; P < 0.01). Chemical restraint did not cause significant changes in IOP. Intraocular pressure can be measured with both types of rebound tonometers in White's tree frogs, but time of day and manual restraint can affect IOP values.