Quantitative measurements of apoptosis using the ACEA NovoCyte Flow Cytometer

In order to demonstrate the flexibility and verstality of the NovoCyte system for assessment of apoptotic events, we show three different assays that can measure apoptosis in response to staurosporine, a broad kinase inhibitor known to induce apoptosis in many cell types.

The first assay uses Annexin V to bind exposed PS on cell membranes for labeling early apoptotic cells and the DNA binding dye, 7-AAD, for labeling late apoptotic cells. 

The second assay makes use of the fluorescent dye JC-1, to measure the loss of mitochondrial membrane potential in early apoptotic cells. JC-1 is cell permeable and aggregates inside mitochondria when membrane potential is maintained, emitting fluorescence in the PE channel.

The third assay quantifies caspase activity. Probe molecules containing a cleavage site specic for caspase 3 & 7 are added to cells. When cleaved by these activated caspases in early apoptotic cells, the probe molecules enter the nucleus, bind to DNA, and fluoresce. This enables the direct readout of activated caspases in cells.

Read the application note.