Amino acid substituted-cresyl violet, synthetic fluorogenic substrates for the analysis of agents in individual in vivo cells or tissue

摘要

The present invention concerns a method to detect the presence of an enzyme in in vivo or in vitro tissue or cell, which method comprises:;(a) obtaining a tissue or cell sample to be analyzed;;(b) contacting the tissue or cell sample with a substrate of the structure selected from the group consisting of: ;X=H or one or more natural or synthetic amino acids with or without amino blocking groups,;Y=H or 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, with the proviso that at least one of X or Y is at least one amino acid;;(c) when an enzyme is present in the tissue or cell sample which degrades X, Y and combinations thereof, fluorescent cresyl violet is released in the tissue sample producing a color change;;(d) detecting the presence and amount of the enzyme present by the detection and quantification of the fluorescence produced; and;(e) optionally comparing the fluorescence to a pre-calibrated fluorescence scale to quantify the fluorescence present. A diagnostic kit for use and a method to prepare amino acid cresyl violet derivatives are described.