Immunofluorescence was utilized to label neurofilaments and astrocytes in a thin section of rat cerebellum tissue. First, the specimen was fixed, permeabilized, blocked with 10-percent normal goat serum, and treated with a cocktail of chicken anti-NF-H (heavy chain neurofilament subunits) and rabbit anti-GFAP (glial fibrillary acidic protein) primary antibodies. Then, to visualize the primary targets, the cerebellum section was treated with goat anti-chicken and anti-rabbit secondary antibodies (IgG) conjugated to Alexa Fluor 488 and Alexa Fluor 568, respectively. Finally, DRAQ5 (pseudocolored cyan) was employed to counterstain cell nuclei. 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.