Fig 11. Analysis of Lysosomal Enzyme Activity Using Alternative Fluorescent Substrates.

AG06173 human fibroblast cells were stained with 10 uM of an esterase substrate with an alternative morpholino based targeting group (6) (C), 10uM of the resorufin based β-glucosidase substrate (9) (B), or 10uM of the resorufin based esterase substrate (10) (A) followed by incubation at 37°C in 5% CO₂ atmosphere for 16 hours. The cells were then washed with pre-warmed PBS followed by addition of Opti-Klear^™ Live Cell Imaging Buffer containing 10ug/ul Hoechst 33342). Cells were imaged using an AMG EVOS Auto FL Fluorescence Microscope equipped with appropriate lightcubes and 40X air objective.