Purpose: To assess the validity of and compare applanation and rebound tonometry readings of intraocular pressure in alert normal chicks from ages 3 to 45 days. Methods: Intraocular pressures (IOPs) were measured weekly in awake White Leghorn chicks, from ages 3-45 days (n = 22-30 per age group), with both applanation Tono-Pen and rebound TonoLab tonometers. Three repeated measurements on individual eyes were used to derive variance data for both instruments at each age. Calibration curves were also derived for each instrument and each age, weekly from ages 10-45 days (n = 3-4 per age group), from in situ manometry data collected over IOP settings of 0 to 100 mmHg in 5 mmHg steps in cannulated eyes. Results: The TonoLab showed less within measurement variability, but more variability with age, than the Tono-Pen. The coefficient of variation ranged from 3.8-8.3% for the TonoLab, compared to 11.0-19.7% for the Tono-Pen across all ages. For the youngest, 3 day-old chicks, mean IOPs recorded with the Tono-Pen and TonoLab were not significantly different (17.0 ± 5.6 and 15.2 ± 3.7 mmHg, respectively, P = .27). However, with increasing age, IOP readings significantly increased for the TonoLab (P < .001), whereas Tono-Pen readings did not. Compared to manometry settings, the Tono-Pen tended to underestimate IOPs while the TonoLab overestimated IOPs over the range 20-60 mmHg, saturating thereafter; there were also age-dependent differences for the TonoLab. Conclusions: Both the Tono-Pen and TonoLab gave IOP readings that differed from manometry settings in normal young chicks over some or all of the ages tested. These results reinforce the importance of calibrating clinical tonometers in animal studies involving IOP as a key variable.