BACKGROUND: In order to determine normal outflow facility and aqueous humor production values in Lewis rats, we established methods of measuring aqueous humor dynamics in this animal. METHODS: Outflow facility was determined using anterior chamber infusion with constant pressure. Aqueous humor production was determined by a technique of dilution with FITC-albumin. RESULTS: The mean outflow facility was 0.044 +/- 0.01 microliter/min/mmHg. Rats weighing less than 300 g had lower values than did rats weighing more than 300 g (0.034 +/- 0.006 microliter/min/mmHg vs 0.050 +/- 0.015 microliter/min/mmHg, P = 0.009). The mean aqueous humor production was 0.350 +/- 0.110 microliter/min. The turnover rate of aqueous humor production per minute was 2.23%. CONCLUSIONS: Anterior chamber infusion with constant pressure and the FITC-albumin method allow measurement of the outflow facility and aqueous humor production in Lewis rats. These methods may be useful in assessing aqueous humor dynamics when rats are used as a glaucoma model.