The continuous flow of the aqueous humor through the posterior and anterior chambers in the eye into Schlemm's canal is vital for the maintenance of a constant intraocular pressure. If the intraocular pressure rises above the normal range, several problems start appearing insidiously; for example, gradual visual field defects develop, leading ultimately to glaucoma. The determination of the flow rate of the aqueous humor is here performed by identifying a mathematical model after the free fluorescein concentration in the blood and in the anterior chamber of the eye has been measured. Experimental data obtained on rabbits are used in the analysis, and the flow rate of the aqueous humor in rabbit eyes is computed. This study demonstrates the feasibility of the approach in a clinical environment.