The intermediate filament protein vimentin was targeted in the rat brain tissue section shown above, which features the hypothalamus and the third ventricle, with mouse anti-vimentin monoclonal antibodies followed by goat anti-mouse secondary antibodies conjugated to Alexa Fluor 488. In addition, glial fibrillary acidic protein was immunofluorescently labeled with rabbit anti-GFAP primary antibodies visualized with goat anti-rabbit secondaries conjugated to Alexa Fluor 568. DRAQ5 (pseudocolored cyan) was subsequently employed as a nuclear counterstain. Images were recorded with a 20x objective using a zoom factor of 1.6 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.