Aqueous humour does not drain uniformly through the trabecular meshwork (TM), but rather follows non-uniform or “segmental” routes. In this study, we examined whether segmental outflow patterns in the TM change over time in living mice and whether such changes are affected by age. Segmental outflow patterns were labelled by constant-pressure infusion of fluorescent tracer microparticles into the anterior chamber of anesthetised C57BL/6J mice at 3 or 8 months of age. Two different tracer colours were infused at separate time points with an interval of Δt = 0, 2, 7 or 14 days. In a separate experiment, one tracer was infusedin vivowhile the second tracer was infusedex vivoafter 2 days. The spatial relationship between the two tracer patterns was analysed using the Pearson’s correlation coefficient,r. In 3-month-old mice, there was a time-dependent decay inr, which was near unity at Δt = 0 and near zero at Δt = 14 days. In 8-month-old mice,rremained elevated for 14 days. Segmental outflow patterns measured in young miceex vivowere not significantly different from those measuredin vivoafter accounting for the expected changes over 2 days. Therefore, segmental outflow patterns are not static in the TM but redistribute over time, achieving near complete loss of correlation by 2 weeks in young healthy mice. There is an age-related decline in the rate at which segmental outflow patterns redistribute in the TM. Further research is needed to understand the dynamic factors controlling segmental outflow.