Kinetic Quantitation of Apoptosis and Necrosis

Apoptosis, or programmed cell death, refers to a series of biochemical processes in the cell leading to distinct morphological changes. These changes can be assessed using fluorescent probes or contrast-enhancing microscopy modes such as phase contrast. Examples of fluorescent probes include those that target phosphatidylserine. This target becomes exposed to extracellular reagents in early stage apoptosis (i.e. pSIVA-IANBD), and is detected using fluorometric substrates of or antibodies to caspases (i.e. Caspase 3/7) that generate fluorescence with caspase activation. Also, the target is exposed with probes that measure chromatin condensation. The Agilent BioTek Cytation cell imaging multimode reader and Agilent BioTek Lionheart FX automated microscope can measure apoptosis by using fluorescent probes. They can also quantify apoptosis by label-free methods relying on phase contrast microscopy.