Transmission of Chagas' disease by transfusion of blood containing Trypanosoma cruzi has often been reported, and gentian violet, a triarylmethane dye, is widely used by blood banks in attempts to eliminate such transmission. In a study of intact trypanosomes, gentian violet was found to undergo a one-electron reduction to produce a carbon-centered free radical as demonstrated by electron spin resonance spectroscopy. Either reduced nicotinamide adenine dinucleotide or the reduced dinucleotide phosphate could serve as a source of reducing equivalents for the production of this free radical by homogenates of Trypanosoma cruzi. The formation of this free radical, and the trypanocidal action of gentian violet, were enhanced by light. The enhanced free radical formation may be the basic cause of the selective toxicity of gentian violet to Trypanosoma cruzi.