In a triple immunofluorescence experiment, the horizontal thin section of rat brain presented in this confocal image was fixed, permeabilized, blocked with 10-percent normal goat serum, and then treated with a cocktail of mouse anti-vimentin (intermediate filaments), rabbit anti-GFAP (astroglia), and chicken anti-MAP2 (neuronal dendrites and perikarya) primary antibodies followed by goat anti-mouse, anti-rabbit, and anti-chicken secondary antibodies (IgG) conjugated to Alexa Fluor 647 (pseudocolored magenta), Alexa Fluor 568, and Alexa Fluor 488, respectively. Subsequently, cell nuclei were counterstained with Hoechst 33342, a nucleic acid probe that fluoresces blue when bound to dsDNA. Images were recorded with a 20x objective using a zoom factor of 1.5 and sequential scanning with the 405-nanometer spectral line of a blue diode laser, the 488-nanometer line of an argon-ion laser, the 543-nanometer line from a green helium-neon laser, and the 633-nanometer line of a red helium-neon laser. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles unless otherwise noted above.