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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 tahr ovary 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 cyanine monomer (TO-PRO-3) reagent.
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