Targeting Neurofilaments and Astroglia in Rat Brain Sections
A rat brain sagittal section was immunofluorescently labeled for phosphorylated neurofilaments with mouse anti-NF-P monoclonal 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 with rabbit anti-GFAP antibodies visualized with goat anti-rabbit secondaries conjugated to Alexa Fluor 568. A dsDNA probe, Hoechst 33342, was used to counterstain cell nuclei. 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.