| Over the past four decades, Multielectrode Array (MEA) devices have played a major role in electrophysiology by providing a simpler solution to simultaneous multisite chronic extracellular recording: in vivo and in vitro. While a wide range of devices have been developed, almost all of them are limited to culturing and recording from one cell type, in vitro; and tissue surfaces, in vivo and in vitro. Most tissues are formed by different cell types that interact to maintain tissue function, like the heart which is composed mainly of cardio-myocytes and fibroblasts. Direct recording from such organs usually employs plunge-type electrodes which induce tissue damage and require better handling for sustenance. To better understand the functioning of such tissues, it is imperative to utilize recording systems that allow interactions between two or more cell types and at the same time sustain cultures with controlled cell number and distribution.;In this thesis, the design, fabrication process, and characterization of an MEA device called the PerFlexMEA (Perforated Flexible MEA) is presented. It enables the generation and sustenance of a preparation with two cell types while recording their electrical activity. PerFlexMEA was developed using a thin (9microm) perforated Polycarbonate Track Etch (PCTE) membrane (3microm diam. pores, 200,000 pores/cm2) as substrate where cells can be cultured on both sides, allowing gap junction formation across the membrane via the pores. Cell number and distribution can be controlled on either side. The PerFlexMEA comprises a 4 x 5 array of square gold electrodes, each measuring 50 microm x 50 microm spaced 500 microm apart. Parylene was patterned to insulate the leads (50 microm thick) connecting the recording electrodes to the contact pads. A coin-shaped device was designed to house the PerFlexMEA and to insulate its cell culture zone (wet) from contact pads (dry). Cardiomyocytes, isolated from neonatal mice were plated on the recording side of PerFlexMEA and electrical activity was recorded at a signal to noise ratio of 8.6 and peak to peak voltage of 200 microV. |
