Development of Patch-Clamp Chips for Mammalian Cell Applications

Abstract

We have previously described the designs of two planar patch-clamp neurochips and their application to the electrophysiological study of molluscan neurons cultured on-chip. Neuron attachment and growth over apertures on the neurochip surface permitted the acquisition of whole-cell patch-clamp recordings. To broaden the application of these neurochips from molluscan to mammalian neurons, we conducted a study of cell-to-aperture interaction to optimize conditions for these smaller, more fragile cells. For this purpose, we designed a “sieve” chip having multiple apertures on its surface. Random growth of rat cortical neurons resulted in a 32% (n = 324) probability of cell growth over 2 μm diameter apertures; larger diameters resulted in growth through the aperture. Based on these findings, single-aperture neurochips were fabricated having 2 μm diameter aperture and preliminary electrophysiological recordings from cortical cultures at 14 DIV are presented. The implications of this study for the next-generation neurochips are discussed.