Neurofilaments, which are found specifically in neurons and are especially prominent in the axons of the cells, are typically composed of light (NF-L), medium (NF-M), and heavy (NF-H) chain subunits. In the dentate gyrus tissue section presented above, the heavy chain neurofilament subunits were targeted with chicken anti-NF-H antibodies followed by goat anti-chicken secondary antibodies conjugated to Alexa Fluor 568. The calcium-binding protein calbindin was simultaneously targeted in the specimen with mouse anti-calbindin antibodies visualized with goat anti-mouse secondary antibodies conjugated to Alexa Fluor 488. Nuclear DNA was subsequently labeled with the far-red fluorescent dye DRAQ5 (pseudocolored blue). Images were recorded with a 20x objective using a zoom factor of 1.2 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.