Neurofilaments are specialized intermediate filaments only present in neurons that are usually composed of a combination of three proteins, known as NF-L, NF-M, and NF-H. These filaments were immunofluorescently labeled in this thin section of rat cerebellar tissue with chicken anti-NF-H antibodies followed by goat anti-chicken secondary antibodies conjugated to Alexa Fluor 488. In addition, glial fibrillary acidic protein, which is an intermediate filament protein found in the central nervous system in certain types of astroglia and neural stem cells, was targeted with rabbit anti-GFAP antibodies visualized with secondary antibodies (goat anti-rabbit) conjugated to Alexa Fluor 568. Cell nuclei were counterstained with DRAQ5 (pseudocolored blue). Images were recorded with a 10x objective using a zoom factor of 1.3 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. |