Real-Time Detection of Apoptosis Using the Incucyte ZOOM® Live-Cell Imaging System

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Essen Bioscience

Time-lapse movie of A549 adenocarcina cells apoptosis following treatment with TNFα (7.5 ng/mL) and cycloheximide (5 µg/mL). Apoptosis was detected in real time using the mix-and-read IncuCyte™ Caspase-3/7 Reagent (a DEVD-linked DNA binding fluoroprobe). When added to growth medium, the IncuCyte™ Caspase-3/7 Reagent is an inert, non-fluorescent substrate that freely crosses the cell membrane and can be cleaved by activated caspase-3/7 in apoptotic cells resulting in the release of a DNA binding green fluorescent label. Apoptotic cells are identified and quantified over time using the IncuCyte ZOOM® system to detect the appearance of green-labelled nuclei. High definition phase contrast images provide an additional qualitative validation of apoptotic cell death based on morphology (e.g. cell shrinkage, membrane blebbing, nuclear condensation).