GeLu Cells with MitoTracker Red CMXRos, BODIPY FL, and TO-PRO-3

A triple fluorophore combination of MitoTracker Red CMXRos, BODIPY FL conjugated to phallacidin, and TO-PRO-3 was used to label an adherent log phase culture of Mongolian gerbil lung cells for mitochondria, the F-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, permeabilized, and blocked with bovine serum albumen. The cells were subsequently labeled with the conjugated phallacidin and counterstained with the cyanine monomer (TO-PRO-3) reagent.