Triple Staining of Ileum Section with Alexa Fluor 568, Oregon Green 488, and Hoechst 33342

A rat ileum tissue section (shown above) was stained with Oregon Green 488 conjugated to wheat germ agglutinin, a plant-derived lectin that targets the Golgi apparatus, as well as Alexa Fluor 568 conjugated to phalloidin for cytoskeletal actin. Cell nuclei were labeled 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.