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
References (20)
20 references, click to expand
-
A single cell electroporation chip.
Lab Chip. 2005 Jan;5(1):38-43
PMID: 15616738
-
Patch clamp techniques for studying ionic channels in excitable membranes.
Annu Rev Physiol. 1984;46:455-72
PMID: 6143532
-
CYTOCENTERING: a novel technique enabling automated cell-by-cell patch clamping with the CYTOPATCH chip.
Receptors Channels. 2003;9(1):59-66
PMID: 12825299
-
Whole cell patch clamp recording performed on a planar glass chip.
Biophys J. 2002 Jun;82(6):3056-62
PMID: 12023228
-
Microfluidic large-scale integration.
Science. 2002 Oct 18;298(5593):580-4
PMID: 12351675
-
An air-molding technique for fabricating PDMS planar patch-clamp electrodes.
Pflugers Arch. 2005 Mar;449(6):564-72
PMID: 15578213
-
Micromolded PDMS planar electrode allows patch clamp electrical recordings from cells.
Biosens Bioelectron. 2002 Jun;17(6-7):597-604
PMID: 11959483
-
Effect of external pH on activation of the Kv1.5 potassium channel.
Biophys J. 2003 Jan;84(1):195-204
PMID: 12524275
-
Control of ion channel expression for patch clamp recordings using an inducible expression system in mammalian cell lines.
BMC Neurosci. 2003 Jul 2;4:15
PMID: 12839626
-
Fluorescence imaging of electrically stimulated cells.
J Biomol Screen. 2003 Dec;8(6):660-7
PMID: 14711391
-
Flux assays in high throughput screening of ion channels in drug discovery.
Assay Drug Dev Technol. 2003 Oct;1(5):709-17
PMID: 15090243
-
Upscaling and automation of electrophysiology: toward high throughput screening in ion channel drug discovery.
Receptors Channels. 2003;9(1):49-58
PMID: 12825298
-
Emerging trends in high-throughput screening.
Curr Opin Pharmacol. 2003 Oct;3(5):522-9
PMID: 14559099
-
Screening for content--the evolution of high throughput.
Nat Biotechnol. 2003 Aug;21(8):859-64
PMID: 12894197
-
Patch clamping by numbers.
Drug Discov Today. 2004 May 15;9(10):434-41
PMID: 15109948
-
Ion-Ion interactions at the selectivity filter. Evidence from K(+)-dependent modulation of tetraethylammonium efficacy in Kv2.1 potassium channels.
J Gen Physiol. 2000 Apr;115(4):509-18
PMID: 10736316
-
Ion-channel assay technologies: quo vadis?
Drug Discov Today. 2001 Dec 15;6(24):1278-1287
PMID: 11738970
-
Inactivation of the sodium channel. I. Sodium current experiments.
J Gen Physiol. 1977 Nov;70(5):549-66
PMID: 591911
-
Patch clamping moves to chips.
Anal Chem. 2004 Sep 1;76(17):327A-330A
PMID: 15452958
-
A benchmark study with sealchip planar patch-clamp technology.
Assay Drug Dev Technol. 2003 Oct;1(5):675-84
PMID: 15090240