Mitochondrial assays flow cytometry assay

As with flow cytometry, multiparameter apoptosis assays may also be performed by confocal laser scanning microscopy (CLSM). Using the approach similar to that detailed above for flow cytometry, we have examined NADPH content, mitochondrial membrane potential (CMX Rosamine fluorescence), and mitochondrial mass (Mitotracker Green), by CLSM. Figure 3 shows an example of a typical multiparameter assay performed by confocal microscopy. 

Christensen, M.E., Jansen, E.S., Sanchez, W., Waterhouse, N.J., 2013. Flow cytometry based assays for the measurement of apoptosis-associated mitochondrial membrane depolarisation and cytochrome c release. Methods 61, 138-145. 

Rottenberg, H. Wu, S. Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. Biochim. Biophys. Acta 1998, 1404, 393-404. 

At the end of treatment, HL-60 cells were stained with 40 nM 3,3 -dihexyloxacarbocyanine (DiOC6 (3), Molecular Probes, Eugene, OR) for 15 min at 37 C. Mitochondrial transmembrane potential was measured by flow cytometry. At least 10,000 cells were collected in each sample. Generation of ROS was monitored with 20 pM 2, 7 -dichlorofluorescein diacetate (DCFH-DA) (Molecular Probes, Eugene, OR) staining for 1 h at 37 C and assayed by flow cytometry. 

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