Retinal Cell Type Identification

In order to identify different retinal cell types, specific immunofluorescence antibodies were applied (n = 5–9 eyes/group; 6 sections/animal, Table 1) [64 (link)]. Briefly, retinal cross sections were blocked with a solution containing 10–20% donkey, 2–3% bovine serum albumin, and/or goat serum and 0.1% Triton-X in PBS. The sections were incubated with primary antibodies at room temperature overnight. Incubation using corresponding secondary antibodies was performed for 1 h on the next day. For the detection of apoptotic cells, a commercially available TUNEL Kit (In Situ Cell Death Detection Kit; Roche; Sigma-Aldrich, St. Louis, MO, USA) was used according to the manufacturer’s instructions. Nuclear staining with 4′,6 diamidino-2-phenylindole (DAPI, Serva Electrophoresis, Heidelberg, Germany) was included to facilitate orientation on the slides. Negative controls were performed for each stain by using secondary antibodies only.