Embryonic Rat Thoracic Aorta Medial Layer Myoblast Cells (A-10 Line)

A-10 Cells with MitoTracker Deep Red 633, Alexa Fluor 488, and SYTOX Orange

A triple fluorophore combination of MitoTracker Deep Red 633, Alexa Fluor 488 conjugated to phalloidin, and SYTOX Orange was used to label an adherent log phase culture of A-10 cells for mitochondria, the filamentous actin network, and nuclear DNA, respectively. The cells were first treated with MitoTracker Deep Red 633 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 phalloidin and counterstained with the SYTOX reagent.