Hæmolysis by brilliant-green and serum

Abstract

The paper deals with the nature of the sensitization of cells with brilliant-green and the kinetics of the hemolysis which occurs upon the addition of serum. The effect of adding brilliant-green to a standard suspension of cells, the extraction of the dye from the cells and from hemoglobin solutions, the fading of the color of the dye under certain conditions, the effect of temp. and the microscopic and spectroscopic appearance of the cells are discussed. Figures and graphs showing the quantitative effect of varying the concentration of the dye and serum on the time of hemolysis are given, together with the formulae for the velocity of the reactions. These considerations support the following conception of the mechanism of hemolysis in these systems. "In the sensitization process, a compound is formed between a protein component of the red cell membrane and either the brilliant-green itself or its leuco-base.[long dash]On the subsequent addition of serum, the serum proteins react with the combined dye in the system to form a hemolysin, which can react with the protein component referred to, and which thus brings about lysis.[long dash]This hypothesis contains 2 essentials, the 1st being that the hemolytic reaction is really composed of 2 consecutive reactions, the 2nd being that the new lysin formed by the combination of the serum proteins and the dye acts upon the cell component to which the dye is united.".