A coronal thin section of rat cerebellum was immunofluorescently labeled for heavy chain neurofilament subunits with chicken anti-NF-H antibodies followed by goat anti-chicken secondary antibodies conjugated to Alexa Fluor 488 (green fluorescence). Neurofilaments are specialized intermediate filaments solely found in neurons, especially in their axons. In addition, glial fibrillary acidic protein, which is expressed in various astroglia and neural stem cells, was targeted with rabbit anti-GFAP antibodies visualized with goat anti-rabbit secondary antibodies conjugated to Alexa Fluor 568 (red fluorescence). Cell nuclei were counterstained with DRAQ5 (pseudocolored blue), a far-red fluorescent DNA probe. Images were recorded with a 10x 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. |