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The development of new fluorogenic reactions for specific chemical groups is of increasing interest in cytochemistry. We describe the application of the nonfluorescent compound isatin on methanol-fixed blood smears. When treated with 0.1% isatin in absolute ethanol or acetone for 30 min and observed under violet-blue exciting light, eosinophil leucocyte granules show a bright green fluorescence. This fluorogenic reaction is abolished after blocking of tryptophan residues by performic acid oxidation. Spectrofluorometric studies with amino acids in vitro reveal that isatin forms a unique and specific fluorescent product with tryptophan.

A specific fluorogenic reaction for tryptophan residues using isatin in organic solvents.