Glial fibrillary acidic protein (GFAP) and vimentin are both members of the class III intermediate filament protein family that were visualized in the rat hypothalamus section illustrated above, which prominently features the third ventricle. Before being treated with a cocktail of rabbit anti-GFAP and mouse anti-vimentin primary antibodies, the specimen was fixed, permeabilized, and blocked with 10-percent normal goat serum. The primary targets were visualized with goat anti-rabbit and anti-mouse secondary antibodies conjugated to Alexa Fluor 568 and Alexa Fluor 488, respectively. Cell nuclei were detected with DRAQ5 (pseudocolored blue), a far-red fluorescent DNA probe. Images were recorded with a 20x objective using a zoom factor of 1.4 and sequential scanning with the 488-nanometer spectral 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.