Assessment of cell phenotype (triple Fluorescent IHC) was conducted on a laser scanning confocal microscope (Leica TCS SP5; Wetzlar, Germany) with a 63x oil lens. Fifty BrdU+ cells were sampled per dentate gyrus across a minimum of five tissue sections. Z plane image stacks (1024×1024 pixels) were taken at 0.8 μm for each cell analyzed. Z-stacks were reconstructed, viewed and analyzed for colabeling within the Leica software similar to that previously described (Nixon et al., 2008).
