Development of Mechanostimulated Patch-Clamp System for Cellular Physiological Study

摘要

Mechanosensitive ion channels play important roles for sensing and responding to the mechanical stimuli signals in living life. Here we report the development of a mechanostimulated patch-clamp system for simultaneous recording of external stimuli and acquisition of cellular physiological responses. This system integrates a custom-designed planar patch-clamp system with a robot-assisted atomic force microscope (AFM) system. The former, with a microfluidic channel, can realize not only recording electrical signals but also exchanging intracellular solution; while the latter, enhanced by robotic techniques (local scan force feedback, augmented reality vision feedback), can generate force stimuli with controllable patterns and magnitudes under the operator's real-time monitoring. To verify the performance of the developed system, we first measured the whole-cell current of the voltage-gated potassium ion channel Kv1.1 expressed on Human Embryonic Kidney (HEK293) cells and then recorded the mechanosensitive ion channel current in a mouse neuroblastoma cell line (Neuro2 A) in the whole-cell configuration during the AFM indenting on the membrane surface; finally, confirmed the ability to exchange intracellular solution by delivering propidium iodide into the captured cell through intracellular solution. The results prove the effectiveness of the system.