In Situ Time-Controllable Chemical Plasma Membrane Injury by Microfluidic Probe Reveals Self-Repair Ability of Single Cells

The loss of plasma membrane integrity poses a serious threat to cells, and the plasma membrane repair mechanism is vital to cell survival. To investigate the plasma membrane repair mechanism, a method for causing controllable plasma membrane injury on single cells and the visualization of the plasma membrane repair process are proposed. A microregion with harmful chemicals is created by open microfluidic probe, which is applied to target single cells for injury. The injury time is precisely controlled by simply moving the target cells in and out of the microregion. Wound-repair process caused by injury is analyzed by fluorescence imaging in real time, revealing the repair kinetics at the single cell level. Using this method, the effect of external conditions (Ca 2+ , Mg 2+ , glucose) on plasma membrane repair in single cells is studied and the dynamics of intracellular species (Ca 2+ ) is monitored. Meanwhile, cellular heterogeneity of plasma membrane repair is revealed. This method provides a precise way to induce chemical plasma membrane injury at the single-cell level, which is a promising tool to study the mechanism of plasma membrane repair.