A novel silicon patch-clamp chip permits high-fidelity recording of ion channel activity from functionally defined neurons.
We report on a simple and high-yield manufacturing process for silicon planar patch-clamp chips, which allow low capacitance and series resistance from individually identified cultured neurons. Apertures are etched in a high-quality silicon nitride film on a silicon wafer; wells are opened on the backside of the wafer by wet etching and passivated by a thick deposited silicon dioxide film to reduce the capacitance of the chip and to facilitate the formation of a high-impedance cell to aperture seal. The chip surface is suitable for culture of neurons over a small orifice in the substrate with minimal leak current. Collectively, these features enable high-fidelity electrophysiological recording of transmembrane currents resulting from ion channel activity in cultured neurons. Using cultured Lymnaea neurons we demonstrate whole-cell current recordings obtained from a voltage-clamp stimulation protocol, and in current-clamp mode we report action potentials stimulated by membrane depolarization steps. Despite the relatively large size of these neurons, good temporal and spatial control of cell membrane voltage was evident. To our knowledge this is the first report of recording of ion channel activity and action potentials from neurons cultured directly on a planar patch-clamp chip. This interrogation platform has enormous potential as a novel tool to readily provide high-information content during pharmaceutical assays to investigate in vitro models of disease, as well as neuronal physiology and synaptic plasticity.
Py
Christophe
C
Institute for Microstructural Sciences, National Research Council of Canada, 1200 Montreal Road, Ottawa, Ontario, Canada. christophe.py@nrc.ca
Denhoff
Mike W
MW
Martina
Marzia
M
Monette
Robert
R
Comas
Tanya
T
Ahuja
Tarun
T
Martinez
Dolores
D
Wingar
Simon
S
Caballero
Juan
J
Laframboise
Sylvain
S
Mielke
John
J
Bogdanov
Alexei
A
Luk
Collin
C
Syed
Naweed
N
Mealing
Geoff
G
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D002478:Cells, Cultured; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D025802:Electric Capacitance; D007473:Ion Channels / Q000187:drug effects*; D008195:Lymnaea; D008564:Membrane Potentials / Q000187:drug effects; D009474:Neurons / Q000187:drug effects*; D018408:Patch-Clamp Techniques / Q000379:methods*; D012825:Silicon
D007473:Ion Channels; D012825:Silicon