<p>Accurate intraocular pressure (IOP) measurement is fundamental for both clinical ophthalmology and experimental research. Study evaluated the performance of iFalcon^TMV100 rebound tonometer in four enucleated rabbit eyes. We employed a stepwise pressure-increasing protocol to generate a physiologically relevant IOP range (5–55 mmHg). At each target pressure, measurements were taken concurrently with the tonometer (using the rabbit-specific mode) and the reference sensor. We used linear regression and Bland-Altman analysis to assess agreement across the entire dataset, supplemented by a segmented analysis of normal and elevated IOP ranges. Overall linear regression revealed a strong correlation between the iFalcon^TMV100 Vet and the reference sensor(R² = 0.9712, <i>p</i> &lt; 0.001); however, interval-dependent bias was present. The positive intercept (0.8951) suggested a tendency for the tonometer to overestimate IOP at low measurement range, while slope &lt; 1 suggested systematic underestimation at higher pressures. Bland-Altman analysis further confirmed this systematic underestimation bias at central corneal position. The iFalcon^TMV100 rebound tonometer showed strong agreement with direct sensor measurements across a broad pressure range (5–55 mmHg) in an ex vivo rabbit model, supporting its potential utility for IOP assessment in this setting, with limitations noted in extreme pressure ranges.</p>