Fig 1: Characterization of Purified RGCs Isolated from Adult Mice. (a–d) Brightfield images (10x) were taken following each panning step during the isolation of adult RGCs from mice: (a) BSL-1 (fibroblasts, endothelial cells, macrophages and microglia), (b) CD11b/c (macrophages and microglia), (c) VC1.1 (amacrine cells), and (d) Thy1.2 (RGCs; scale bar = 250 µm). As expected, Thy1.2+ cells (inset, d) were larger than BSL-1+ (inset, a), CD11b/c+ (inset, b), and VC1.1+ (inset, c) cells (scale bar = 50 µm). (e) Bar graph showing the distribution of the diameter (µm) of the Thy1.2+ cells (n = 2,759 cells). The average cell diameter was 11.3 ± 1.8 µm. (f) 4-step immunopanning of RGCs average yield of 46,718 ± 2,176 cells per retina with an 86.7 ± 1.2% viability per isolation (n = 21). Immunostaining of the isolated cells against RGC Markers, TUJ1 (h) and RBPMS (i), as well as a nuclear stain, DAPI (g; scale bar = 50 µm). Merged image (j) identified 88.1 ± 4.2% of DAPI positive cells to be RBPMS positive. (k) Semi-quantitative q-PCR analysis identified a significant (p < 0.0001) increase in expression of Pou4f2 (RGC marker) and significant decrease (p < 0.0001) in the expression of Rhodopsin (Rho; Photoreceptor gene marker) in the purified RGCs samples compared to the whole retina.