Tissue was perfused with 4% paraformaldehyde in PBS, postfixed overnight, sectioned at a thickness of 25 µm,incubated in blocking solution (0/25% lambda carageenen, 1%bovine serum albumin, and 0.5% Triton X-100 in 0.01M PBS) and immunostained using antibodies to the dendritically-localized microtubule protein MAP2 (polyclonal Ab266 from S. Halpain), and the astroglial cytoskeletal protein GFAP (glial fibrillary acidic protein, monoclonal clone G-A-5, Sigma) followed by fluorescently-conjugated secondary antibodies (anti-mouse Alexa 488, 1:400; anti-rabbit Alexa 546, both from Molecular Probes), and incubated in 1:1000 Hoescht33258 to stain nuclei. A multiframe data set of 5x3x20(X,Y,Z) was acquired using the Leica SP5 confocal microscope with line-by-line sequential scanning, a 63X HCX PL APOCHROMAT, 1.32NA, with 1.7 zoom at z-intervals of 0.5 µm, and saved as a tif image stack.