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Micro and Nanosystems

Editor-in-Chief

ISSN (Print): 1876-4029
ISSN (Online): 1876-4037

Development of Patch-Clamp Chips for Mammalian Cell Applications

Author(s): Dolores Martinez, Marzia Martina, Laura Kremer, Robert Monette, Tanya Comas, Danish Salim, Christophe Py, Mike W. Denhoff and Geoff Mealing

Volume 2, Issue 4, 2010

Page: [274 - 279] Pages: 6

DOI: 10.2174/1876402911002040274

Price: $65

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.

Keywords: Mammalian cells, planar patch-clamp, neurochip, whole-cell recordings, apertures, poly(dimethylsiloxane), PDMS, Lymnaea neurons, dissecting microscope, Sieve Chip, polyimide, Plexiglas, Minimal Essential Medium, L-glutamine, Normal Bath Media, Calcein-AM, RH-237, LSM-410 Zeiss, confocal microscope, krypton/argon, North-ern Eclipse Software, Electrophysiology, DIV, scanning electron microscopy, SEM, focused, FIB


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