Targeting F-Actin and Mitochondria in Pl 1 UT Cell Cultures

A triple fluorophore combination of MitoTracker Red CMXRos, BODIPY FL conjugated to phallacidin, and DRAQ5 (pseudocolored cyan) was used to label an adherent log phase culture of raccoon uterus 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 phallotoxin and counterstained with the anthraquinone (DRAQ5) reagent.