The triple fluorophore combination of MitoTracker Orange CMTMRos (pseudocolored yellow), BODIPY FL conjugated to phallacidin, and DRAQ5 was used to label an adherent log phase culture of BPAE cells for mitochondria, the filamentous actin network, and nuclear DNA, respectively. The cells were first treated with the MitoTracker probe in growth medium for one hour, washed and fixed with paraformaldehyde (prepared in growth medium), permeabilized, and blocked with bovine serum albumen. The cells were subsequently labeled with the conjugated phallacidin and counterstained with the anthraquinone (DRAQ5) reagent. Images were recorded with a 60x oil immersion objective using a zoom factor of 1.5 and sequential scanning with the 488-nanometer spectral line of an argon-ion laser, the 543-nanometer line from a green helium-neon laser, and the 633-nanometer line of a red helium-neon laser. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles unless otherwise noted above.