The coronal section of rat cerebellum presented in this confocal image was stained for astrocytes, neurons, and cell nuclei. Primary rabbit antibodies against glial fibrillary acidic protein, which is a member of the class III intermediate filament protein family specifically expressed in astrocytes and certain other stellate cells in the central nervous system, were visualized with goat anti-rabbit secondary antibodies conjugated to Alexa Fluor 568 (red fluorescence emission). Chicken antibodies against heavy chain neurofilament subunits, which are solely expressed in neurons, were simultaneously employed to target neural cells, and these primary targets were visualized with a secondary antibody (goat anti-chicken) conjugated to Alexa Fluor 488 (green emission). To label cell nuclei, the specimen was subsequently treated with DRAQ5 (pseudocolored cyan). Images were recorded with a 20x objective using a zoom factor of 1.0 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. View a larger version of this digital image. |