Immunostaining of Macaque Retinal Cryosections

Dissection and preparation of mouse cochleas were performed as previously described [7 (link)]. Eyes obtained from deeply anaesthetized adult macaques perfused with paraformaldehyde, were dissected and processed as previously described [8 (link)]. Retinal cryosections were incubated in 10% bovine serum albumin in phosphate buffered saline (PBS) for 1 hour, incubated with the appropriate primary antibody overnight at 4°C, rinsed in PBS, incubated with the appropriate secondary antibody for 1 hour at room temperature, and rinsed again in PBS. The primary antibodies used were anti-EPS8 (BD Transduction Laboratories), anti-synaptophysin (MAB368, Millipore), anti-whirlin (G-8, Santa Cruz Biotechnology, Inc) mouse monoclonal antibodies, and anti-EPS8 (M-238, Santa Cruz Biotechnology, Inc) rabbit polyclonal antibody. Secondary antibody was Alexa Fluor 488 goat anti-rabbit IgG. TRITC-phalloidin (Sigma-Aldrich) and DAPI (1 μg/ml; Sigma-Aldrich) were used to label F-actin and cell nuclei, respectively.