Development of Mechanostimulated Patch-Clamp System for Cellular Physiological Study

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.