I want to monitor apoptosis changes in cells in periodic manner. when stains are added it fix the cells in particular state. I would like to monitor apoptosis continuously for 24 hours,or until the programmed cell death cycle get completed. What is the better way to do this?
You can use assay with annexin V-FITC and propidium iodide with allows us to determine early apoptotic (annexin V-FITC positive) and late apoptotic and necrotic cells Using this assay you can not distinguish late apoptotic from necrotic cells
Apoptosis, or programmed cell death refers to a series of biochemical changes in the cell leading to distinct morphological changes that can be assessed using fluorescent probes or contrast-enhancing microscopy modes such as phase contrast. Examples of fluorescent probes include those that target phosphatidylserine (i.e. Annexin V), which becomes exposed to extracellular reagents in early stage apoptosis, fluorometric substrates of or antibodies to caspases (i.e. Caspase 3/7) that generate fluorescence with caspase activation, or probes that measure chromatin condensation. Cytation 5 and Lionheart FX both have the ability to measure apoptosis through the use of fluorescent probes or by label-free methods relying on phase contrast microscopy.
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