Detecting mitochondrial potential
– assessing apoptosis using the NC-3000™
- Easy discrimination between polarized (healthy) cells, depolarized (apoptotic) cells and
necrotic/late apoptotic cells
- Fast automated single cell analysis
- Acquisition and analysis in a simple step
- Standardized results – even with different users
- No calibration required
- PlotManager for superior data presentation
- Automated PDF reports
- Export of data in FCS/ACS formats
Loss of the mitochondrial membrane potential is known to precede apoptosis and chemical-hypoxia-induced necrosis. The lipophilic cationic dye, JC-1, display potential-dependent accumulation in the mitochondria and provides a simple, fluorescent-based method for distinguishing between healthy and apoptotic cells.
In healthy cells, the negative charge established by the intact mitochondrial membrane potential facilitates the accumulation of JC-1 in the mitochondrial matrix.
At high concentrations JC-1 forms aggregates and become red fluorescent. In apoptotic cells the mitochondrial potential collapses and JC-1 localizes to the cytosol in its monomeric green fluorescent form.
Using fluorescence microscopy and image analysis, the NucleoCounter® NC-3000™ system automates detection of cells with collapsed mitochondrial membrane potential. Cells are stained with JC-1 and DAPI.
Cellular JC-1 monomers and aggregates are detected as green and red fluorescence, respectively. Mitochondrial depolarization is revealed as a decrease in the red/green fluorescence intensity ratio. Necrotic and late apoptotic cells are detected as blue fluorescent (DAPI) cells.