Sagittal Rat Brain Section Stained with Alexa Fluor Dyes and a Popular Nuclear Counterstain

Antibodies directed against myelin CNPase detect developing and adult myelin and often serve as markers of oligodendrocytes and Schwann cells. A mouse anti-myelin CNPase antibody followed by a goat anti-mouse secondary antibody conjugated to Alexa Fluor 488 was utilized to immunofluorescently label the sagittal section of rat brain tissue presented in the digital image above. In addition, glial fibrillary acidic protein was targeted in the tissue section by treating the specimen with a rabbit anti-GFAP antibody visualized with a goat anti-rabbit secondary antibody 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.