A protocol to measure lysosomal Zn 2+ release through a genetically encoded Zn 2+ indicator.

Intracellular vesicles such as lysosomes contain micromolar to millimolar concentrations of Zn, and disturbing lysosomal Zn homeostasis via lysosomal Zn release leads to mitochondria damage and consequent lytic cell death. Methods have been developed to image cellular Zn dynamics. Here, we present a protocol using GZnP3, a genetically encoded fluorescent Zn indicator, to assess lysosomal Zn 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).