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

Zinc (Zn(2+)) is a trace element, playing pivotal roles during host-pathogen interactions. Macrophages can sequester Zn(2+) and restrict bioavailability or increase phagolysosomal Zn(2+) to kill pathogens. This method quantifies Zn(2+)-mediated clearance of the human fungal pathogen C. glabrata 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(2+) decreases fungal viability as a function of intracellular Zn(2+) concentrations in macrophages. For complete details on the use and execution of this protocol, please refer to Riedelberger et al. (2020).