The bovine pulmonary artery endothelial (BPAE) cells presented in the digital image above were resident in an adherent culture stained for F-actin with Alexa Fluor 488 conjugated to phalloidin (green fluorescence), and for DNA with the bis-benzimidazole dye Hoechst 33258 (blue fluorescence). In addition, the culture was immunofluorescently labeled with Alexa Fluor 568 conjugated to goat secondary antibodies that target mouse anti-PDI (protein disulfide isomerase) primary antibodies (red fluorescence). The mitochondrial network was simultaneously visualized using MitoTracker Deep Red 633 (pseudocolored yellow). 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 Omega Optical. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.