Quantification of zinc intoxication of Candida glabrata after phagocytosis by primary macrophages.

Zinc (Zn) is a trace element, playing pivotal roles during host-pathogen interactions. Macrophages can sequester Zn and restrict bioavailability or increase phagolysosomal Zn to kill pathogens. This method quantifies Zn-mediated clearance of the human fungal pathogen after phagocytosis by innate immune cells. Double staining with propidium iodide and a zinc-specific fluorescence dye allows for discrimination of live versus dead pathogens inside phagolysosomes. Moreover, elevated phagolysosomal Zn decreases fungal viability as a function of intracellular Zn concentrations in macrophages. For complete details on the use and execution of this protocol, please refer to Riedelberger et al. (2020).