Single-Cell Localized Treatment and ROS Dynamics Monitoring via an Open Electrochemical-Microfluidic Probe

Capturing immediate responses to diverse stimulations in single living cells offers new insights into cellular behaviors and functions. However, integrating precise micro-perturbations with in-situ acquisition of molecular information remains challenging. Here, an open electrochemical-microfluidic probe (OEMP) is developed for single-cell localized treatment and reactive oxygen species (ROS) dynamics monitoring. The OEMP integrates fluidic apertures and functionalized carbon microelectrode in an open interface, which can create a hydrodynamic flow confinement area for precise cell manipulation, followed by real-time ROS evaluation via electrochemical sensing. As a proof of concept, oxidative stress during the ferroptosis process is investigated by stimulating the whole single cell, elucidating the regulatory role of glucose through amperometric parameters. Furthermore, an expanded fluidic configuration can create symmetrical areas within single cells for partial membrane damage and successfully track ROS dynamics involved in potential wound-repair mechanisms. This versatile approach provides a promising way for acquiring in-situ molecular insights and advancing understandings of biological processes from a single-cell perspective.