Visualizing Cochlear Hair Cells

The basilar membrane samples were permeabilized with 2.5% Triton X-100 in 1X PBS for 15 min at room temperature on a shaker. Then, the specimens were washed 3 times with PBS and blocked in 10% goat serum solution for 1 h at room temperature. After washing with PBS three times, cochlear sections were incubated with phalloidin (1:200) for 2 h at room temperature in the dark, counterstained with DAPI for 8 min and washed three times with PBS. The samples were mounted on glass slides in 10 µl anti-fluorescence quenching agent. Hair cells were visualized using an Olympus BX63 fluorescence microscope from the apex to the base of the cochlea, and then the outer hair cells were counted.