Figure 4.

Enhanced tracer deposition in outflow tissues of living mice subjected to netarsudil mesylate treatment. Fluorescent microbeads were loaded into microneedles in the presence or absence of netarsudil mesylate, anterior chambers were cannulated and paired eyes simultaneously perfused at a constant flow rate of 0.167 μl/min. Perfusion was stopped after one hour and the mice were maintained for another hour before being euthanized. Shown are flat mounted anterior segments of C57 and CD1 mouse eyes that were visualized using epifluorescence microscopy (panel A). Summary of comparisons between netarsudil mesylate-treated (NT) and vehicle (0.001% DMSO)-treated groups are shown in panel B. Fluorescence intensity, the width and area of fluorescence in conventional outflow region were automatically quantified. Data represent mean ± S.E.M, N=5 for both strains of mice. *, P < 0.05, **, P < 0.01 compared to their contralateral vehicle control eyes by student t-test. Panels C and D show representative fluorescence intensity and width maps of conventional outflow regions from four quadrants of paired eyes. The variation of mean intensity (of all pixels with identical angles to the center) (panels C) and the variation of width (panels D) are shown with the variation of angle from the center of the circular TM/SC regions (from −180 to 180 degrees). The 0 (180) degree is represented by the positive X-axis.