Localizing Fluorescent Tags to Filamentous Actin and Golgi Complexes in Gastric Tissue
This widefield image of a rat stomach tissue sample was produced by probing the specimen with Alexa Fluor 568, Oregon Green 488, and Hoechst 33342. The Alexa Fluor dye was conjugated to phalloidin, targeting the cytoskeletal F-actin network, and Oregon Green 488 was conjugated to WGA in order to localize a green fluorescent tag to the Golgi complex. The nuclear counterstain Hoechst 33342 was employed to visualize 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.