Patent ID: 6235493
Filing Date: 2001-05-22
Classification: C12Q

Abstract:
A method to detect the presence of an enzyme in living individual cells, which method comprises:(a) obtaining living individual cells to be analyzed;(b) contacting the living individual cells with a substrate of the structure: ##STR5##whereinX is selected from one or more natural or synthetic amino acids with or without amino blocking groups,Y is selected from one or more natural or synthetic amino acids with or without amino blocking groups,wherein X and Y are the same or different and are amino acid sequences of between about 1 to 1,000,000 amino acids wherein each amino acid is the same or a different amino acid;wherein when an enzyme which degrades X or Y is present in the living individual cells,m fluorescent cresyl violet is released producing a color change wherein the color produced remains within the living individual cells;(c) detecting the presence and amount of the enzyme present by the detection and quantification of fluorescence produced; and(d) comparing the fluorescence to a pre-calibrated fluorescence scale to quantify the fluorescence present.