Triple Staining of Stomach Tissue Sample with Alexa Fluor 568, Oregon Green 488, and Hoechst 33342

The cytoskeletal F-actin network was targeted in a sample of the fundus region of a rat stomach (presented above) with phalloidin conjugated to Alexa Fluor 568. Phalloidin is a member of the phallotoxin group of bicyclic peptides isolated from the deadly Amanita phalloides mushroom. The tissue section was also labeled for nuclear DNA with Hoechst 33342 and the Golgi complex with Oregon Green 488 conjugated to wheat germ agglutinin (WGA). 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.