Image processing and analysis of immuno-stained sections

Image acquisition was performed with a laser-scanning confocal imaging system (Zeiss LSM710, 25X, and Leica TCS SP8, 40X) and image quantification was performed with ImageJ software (NIH), and Imaris (Oxford Instruments). DCX + cells and DCX, Ki-67 co-immunostaining were manually quantified in confocal z-stack images (25X lens and 40X lens, respectively) by the 3D reconstruction in Imaris. Specifically, confocal stacks of images of both hemispheres’ dentate gyri were obtained from 3 to 4 rostral sections. Cells were counted manually, and data were expressed as number of cells per section. For neuroblast morphology, high-resolution confocal images stacks of hippocampus from 3 rostral sections were obtained. At least 30 neurons were traced and analyzed using Imaris filaments tool. Apical dendrite length was measured in Imaris.