Live Cell Microscopy and Flow Cytometry to Study Streptolysin S-Mediated Erythrocyte Hemolysis

The ability to induce hemolysis, the rupturing of erythrocytes with the consequent release of their intracellular contents, is a phenotypic hallmark of a number of microbial toxins. Streptococcus pyogenes or Group A Streptococcus (GAS) is a human pathogen responsible for a wide range of diseases from mild pharyngitis to severe conditions such as toxic shock syndrome. GAS produces a powerful hemolytic toxin called streptolysin S (SLS). Herein, we describe a procedure for the preparation of SLS toxin and the use of two complementary approaches, live microscopy and flow cytometry, to study the effects of the SLS toxin on erythrocytes. In addition to providing insights into SLS-mediated hemolysis, these assays have the potential to be modified for the study of other hemolytic toxins and compounds.