Fabrication of Glass Micropipette Device Using Reflow Processes and Its Integration With Microfluidic Channels for Patch Clamp Recording of Cell

Abstract

This paper presents a borosilicate glass micropipette device that has a high-aspect-ratio pore for patch clamp recording applications. The device was fabricated by carrying out glass reflow processes on a silicon substrate. The shape of the micropipette was determined by the width and thickness of the reflowed glass structure, which matches well with the rounded tip of a conventional glass pipette. The diameter of the micropipette pore was first defined by deep reactive ion etching (DRIE) of a silicon pillar, which was positioned at the center of the glass structure. Then, the silicon pillar was removed, and the pore diameter was additionally reduced to the submicron scale by depositing a parylene-C film. The fabricated device was integrated with microfluidic channels for patch clamp recording experiments: microfluidic PDMS layers were assembled on the top and bottom surfaces of the micropipette chip for the medium exchange and the application of negative pressure, respectively. The quality of seal between a cerebral cortex cell of a rat and the glass micropipette was characterized by the amplitude of the current produced by an applied voltage pulse. A negative pressure applied through the bottom microfluidic channel pulled down the cell and increased the seal resistance from 9.82M Omega to 2.06 G Omega. This indicates that the gigaseal was successfully formed around the micropipette pore. In addition, the action potentials of the patched cortical neuron were recorded using the fabricated device.