Frozen sections (4 μm) were cut, air dried, and washed in phosphate-buffered saline (PBS), blocked with 20% goat serum in 0.1% Triton X-100/1% bovine serum albumin (BSA; Sigma-Aldrich, St. Louis, MO) in PBS. For RPE65 single staining, the sections were incubated with the rabbit anti-RPE65 antibody overnight at 4°C. For cone opsin and RPE65 double staining, the sections were incubated with biotin-conjugated anti-RPE65 antibody and an FITC-conjugated anti-bovine medium wave length (MWL) cone opsin antibody. The sections were rinsed several times with PBS and incubated with Texas red–conjugated goat anti-rabbit antibody (for single staining) at a dilution of 1:150 for 30 minutes or Texas red–conjugated goat streptavdin (for double staining; Jackson ImmunoResearch Laboratory, Inc., West Grove, PA). After this, the slides were rinsed in PBS and the nucleus was stained with 4′,6-diamidino-2-phenylindole (DAPI; Sigma-Aldrich, St. Louis, MO). The sections were then mounted in antifade medium (Vectashield; Vector Laboratories, Burlingame, CA) and viewed on a laser scanning confocal microscope (model LSM 510; Carl Zeiss Meditec, Inc., Jena, Germany).