DC microelectrode array for investigating the intracellular ion changes

Abstract

Microelectrode arrays (MEAs) are extensively being used to study the electrical properties of cells. Most of the MEAs use metal electrodes which are in direct contact with the cells. When using DC currents, this leads to undesirable chemical influencing of the cell. Also, metal electrodes are unsuitable for the measuring of constant potentials. A new kind of MEA is developed which replaces the metal electrodes by electrolyte-filled microchannels with Ag/AgCl-electrodes at their ends. The surface of the DCMEA consists of a nanoporous membrane that acts as a homogenous cell substrate, thus avoiding any topographical guidance of the cells. It is adhered to a polydimethylsiloxane layer with four electrode channels embedded in it, using a novel plasma bonding method. A transparent polymer ground plate connects the channels to the silver electrodes as shown in Fig. 1. This MEA allows for the stimulation of the cells with stationary, non-homogenous electric fields, e.g. to simulate the electrical environment near wounds in vitro. It has been proposed in the literature that intracellular ions are involved during cell migration. The DCMEA can be used to simulate in vitro electric fields to investigate intracellular ion changes. By loading cells with ion specific fluorescence dyes, real-time ion kinetic changes can directly be carried out on DCMEA. These studies will be performed by using a time lapse video microscope. In this paper we present the detailed fabrication and testing of the new DCMEA. Results on intracellular ion flows will be presented using this DCMEA.