A protocol to measure lysosomal Zn²⁺ release through a genetically encoded Zn²⁺ indicator

Intracellular vesicles such as lysosomes contain micromolar to millimolar concentrations of Zn²⁺, and disturbing lysosomal Zn²⁺ homeostasis via lysosomal Zn²⁺ release leads to mitochondria damage and consequent lytic cell death. Methods have been developed to image cellular Zn²⁺ dynamics. Here, we present a protocol using GZnP3, a genetically encoded fluorescent Zn²⁺ indicator, to assess lysosomal Zn²⁺ release in cultured cells by fluorescence microscopy imaging. For complete details on the use and execution of this protocol, please refer to Du et al. (2021) or Minckley et al. (2019).