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A membrane-associated FRET sensor for detection of apoptosis

A membrane associated caspase sensor based on Förster Resonance Energy Transfer (FRET) between enhanced cyan fluorescent protein (Mem-ECFP) and yellow fluorescent protein (EYFP) is reported. Upon apoptosis a caspase sensitive amino acid peptide linker (DEVD) between these proteins is cleaved, and pronounced changes of fluorescence spectra and lifetimes are observed. Membrane selective detection of fluorescent proteins in cultivated HeLa cervix carcinoma cells is achieved by total internal reflection fluorescence microscopy (TIRFM) with high sensitivity and resolution.