Brain Tissue Sample Probed with Alexa Fluor 488, Alexa Fluor 568, and Hoechst 33342
The horizontal section of rat brain depicted above was immunofluorescently labeled for neurofilaments with mouse anti-NF-H Non PO4 antibodies followed by goat anti-mouse secondary antibodies conjugated to Alexa Fluor 488. In addition, glial fibrillary acidic protein (expressed in various astroglia and neural stem cells) was targeted with rabbit anti-GFAP antibodies visualized with goat anti-rabbit secondary antibodies conjugated to Alexa Fluor 568. Cell nuclei were labeled with the popular nuclear counterstain 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.