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PMID: 15967996 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Mammalian electrophysiology on a microfluidic platform.

Ionescu-Zanetti C, Shaw RM, Seo J, Jan YN, Jan LY, Lee LP

Abstract

The recent development of automated patch clamp technology has increased the throughput of electrophysiology but at the expense of visual access to the cells being studied. To improve visualization and the control of cell position, we have developed a simple alternative patch clamp technique based on microfluidic junctions between a main chamber and lateral recording capillaries, all fabricated by micromolding of polydimethylsiloxane (PDMS). PDMS substrates eliminate the need for vibration isolation and allow direct cell visualization and manipulation using standard microscopy. Microfluidic integration allows recording capillaries to be arrayed 20 microm apart, for a total chamber volume of <0.5 nl. The geometry of the recording capillaries permits high-quality, stable, whole-cell seals despite the hydrophobicity of the PDMS surface. Using this device, we are able to demonstrate reliable whole-cell recording of mammalian cells on an inexpensive microfluidic platform. Recordings of activation of the voltage-sensitive potassium channel Kv2.1 in mammalian cells compare well with traditional pipette recordings. The results make possible the integration of whole-cell electrophysiology with easily manufactured microfluidic lab-on-a-chip devices.

MeSH Terms
Animals CHO Cells Capillaries/metabolism Cricetinae Dimethylpolysiloxanes/chemistry Electrophysiology/methods Microfluidic Analytical Techniques Microfluidics Patch-Clamp Techniques/methods Potassium Channels, Voltage-Gated/chemistry Shab Potassium Channels Silicones/chemistry Temperature
Chemicals
Dimethylpolysiloxanes Potassium Channels, Voltage-Gated Shab Potassium Channels Silicones baysilon
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Ionescu-Zanetti Cristian
Department of Bioengineering, University of California, Berkeley, CA 94720, USA.
Shaw Robin M
Seo Jeonggi
Jan Yuh-Nung
Jan Lily Y
Lee Luke P
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2005-06-28
Epub
2005-00-20
Pages
9112-7
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC1166618
Subset
IM
Grants
NIMH NIH HHS · R01 MH065334 · United States
NIMH NIH HHS · R37 MH065334 · United States
NIMH NIH HHS · MH65334 · United States
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