The peroxisome organelles present in the rat thoracic aorta fibroblast cell culture illustrated above were immunofluorescently labeled with Rhodamine Red-X conjugated to antibodies directed against peroxisomal membrane protein 70 (PMP 70). Alexa Fluor 488 conjugated to phalloidin, MitoTracker Red CMXRos and Hoechst 33258 were simultaneously used to counterstain the culture, targeting F-actin, mitochondria and DNA, respectively. Images were recorded in grayscale with a Hamamatsu ORCA-AG camera system coupled to an Olympus BX-51 microscope equipped with bandpass emission fluorescence filter optical blocks provided by Semrock. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.