Bovine pulmonary artery endothelial cells (BPAEC) stained with 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, FM 5-95 and Hoechst 33342.
Bovine pulmonary artery endothelial (BPAEC) cells were initially stained with the reactive oxygen species (ROS) indicator, 6-carboxy-2',7'-dichlorodihydrofluorescein diacetate, di(acetoxymethyl ester) (C2938). After a 30-minute incubation, the cells were washed and then incubated simultaneously with FM 5-95 (T23360) and Hoechst 33342 (H1399, H3570, H21492) in phosphate-buffered saline (PBS) for an additional five minutes before washing and mounting in PBS. The red-fluorescent FM 5-95 appears to stain both the plasma membrane and early endosomes; the green-fluorescent, oxidized carboxydichlorofluorescein localizes to the cytoplasm; and the blue-fluorescent Hoechst 33342 dye stains the nucleus.
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