Myelinated Nerve Fibers and Astroglia Visualized in a Sagittal Section of Rat Brain

Myelin was targeted in the sagittal rat brain section presented above with mouse anti-myelin CNPase antibodies followed by goat anti-mouse secondary antibodies conjugated to Alexa Fluor 488. In addition, glial fibrillary acidic protein was targeted in the tissue section by treating the specimen with rabbit anti-GFAP antibodies visualized with goat anti-rabbit secondary antibodies conjugated to Alexa Fluor 568. Nuclear DNA was counterstained with Hoechst 33342. Images were recorded in grayscale with a 12-bit digital camera coupled to a Nikon Eclipse 80i microscope equipped with bandpass emission fluorescence filter optical blocks. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.