Immunocytochemistry of hair cells and quantification of hair cells and neurons
Immunostaining of P7 cochlear whole mounts using the hair cell specific-myosin VII (MyoVIIa) antibody was performed as previously described [2]. Images of labeled organs of Corti were then acquired using a SPOT digital camera (Diagnostic Instruments Inc.) and the number of hair cells was scored from acquired images. To determine the number of hair cells or neurons in vestibular endorgans or sensory ganglia, serial sections were stained with Cresyl Violet and images were similarly acquired and scored. Every other section was scored for each sensory epithelium and every forth section was scored for each ganglion. Only neurons or cells with a clear nucleus and nucleoli were counted and 4–6 samples were counted for each epithelium and ganglion. All data were tested for significance using two-sample Student's t-test with unequal variances.