Protocol for measuring cell cycle Zn²⁺ dynamics using a FRET-based biosensor

The exchangeable Zn²⁺ pool in cells is not static but responds to perturbations as well as fluctuates naturally through the cell cycle. Here, we present a protocol to carry out long-term live-cell imaging of cells expressing a cytosolic Zn²⁺ sensor. We then describe how to track cells using the published pipeline EllipTrack and how to analyze the single-cell traces to determine changes in labile Zn²⁺ in response to perturbation. For complete details on the use and execution of this protocol, please refer to Rakshit and Holtzen et al.¹.