Protocol for Quantifying Zinc Flux in Cultured Cells using Fluorescent Indicators

Zinc (Zn2+) plays a vital role in the functioning of the cell. Cells have influx and efflux zinc transporters to regulate the levels of Zn2+ in the cytoplasm and organellar compartments to maintain homeostasis. We present a protocol to measure changes in cellular zinc concentrations using either a low-affinity membrane permeable or a high-affinity membrane impermeable fluorescent dye. Overall, zinc-specific fluorescent indicators using the assay can reliably detect the Zn2+ flux into or out of cultured cells. For complete details on the use and execution of this protocol, please refer to Sanchez et al. (2019). [Display omitted] •Reliable measure of zinc flux into or out of cultured adherent cells•Amenable for use with fluorescent dyes or genetically encoded zinc indicators•Data and statistical analyses can be done using a standard spreadsheet•Adaptable assay for other metal ions that bind specific fluorescent dyes Zinc (Zn2+) plays a vital role in the functioning of the cell. Cells have influx and efflux zinc transporters to regulate the levels of Zn2+ in the cytoplasm and organellar compartments to maintain homeostasis. We present a protocol to measure changes in cellular zinc concentrations using either a low-affinity membrane permeable or a high-affinity membrane impermeable fluorescent dye. Overall, zinc-specific fluorescent indicators using the assay can reliably detect the Zn2+ flux into or out of cultured cells.