Measuring Ocular Perfusion and Outflow

Ocular perfusion and outflow measurements were done with a system established for the anterior chamber perfusion model [8 (link), 18 (link)–21 ]. Before each use, the system was calibrated with a water column calibration kit. Briefly, anterior segments were cultured at 37 °C and perfused at 4 μl/min with media (Dulbecco’s modified Eagle medium (DMEM); sh30284.02, Fisher Scientific)), 1% FBS (10082-147; Fisher Scientific), 1% antibiotic and antimycotic (15240-062; Fisher Scientific), and a microinfusion pump (70-3007; Harvard Apparatus, Holliston, MA, USA). IOP was measured at 2-min intervals with pressure transducers (Deltran II: DPT-200; Utah Medical Products, Midvale, UT, USA) and recorded (FE224, PL3508/P, MLA1052; ADInstruments, Sydney, Australia; LabChart 7; ADInstruments) [18 (link)]. Baseline IOP was achieved after 48 h of perfusion. The effect of netarsudil on IOP was observed over the subsequent time.