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High-fidelity patch-clamp recordings from neurons cultured on a polymer microchip

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Abstract

We present a polymer microchip capable of monitoring neuronal activity with a fidelity never before obtained on a planar patch-clamp device. Cardio-respiratory neurons Left Pedal Dorsal 1 (LPeD1) from mollusc Lymnaea were cultured on the microchip’s polyimide surface for 2 to 4 hours. Cultured neurons formed high resistance seals (gigaseals) between the cell membrane and the surface surrounding apertures etched in the polyimide. Gigaseal formation was observed without applying external force, such as suction, on neurons. The formation of gigaseals, as well as the low access resistance and shunt capacitance values of the polymer microchip resulted in high-fidelity recordings. On-chip culture of neurons permitted, for the first time on a polymeric patch-clamp device, the recording of high fidelity physiological action potentials. Microfabrication of the hybrid poly(dimethylsiloxane)—polyimide (PDMS-PI) microchip is discussed, including a two-layer PDMS processing technique resulting in minimized shrinking variations.

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Abbreviations

PDMS:

poly(dimethylsiloxane)

PI:

polyimide

LPeD1:

Left Pedal Dorsal 1

R-C:

access resistance – shunt capacitance

SEM:

scanning electron microscope

AFM:

atomic force microscope

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Acknowledgments

The authors wish to gratefully acknowledge the technical support of Allan Horner and Robert Trepanier for Plexiglas packages fabrication, Hue Tran for metallization, Jeff Fraser for scanning electron microscopy imaging and Raluca Movileanu for AFM data. Naweed Syed was supported by the Canadian Institute for Health Research (CIHR) grant. Collin Luk is the recipient of NSERC and Alberta Heritage Foundation for Medical Research (AHFMR) studentships.

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Authors and Affiliations

  1. Institute for Microstructural Sciences, National Research Council of Canada, 1200 Montreal Road, Ottawa, Ontario, K1A0R6, Canada

    Dolores Martinez, Christophe Py & Mike W. Denhoff

  2. Institute for Biological Sciences, National Research Council of Canada, 1200 Montreal Road, Ottawa, Ontario, K1A0R6, Canada

    Marzia Martina, Robert Monette, Tanya Comas & Geoff Mealing

  3. Hotchkiss Brain Institute, University of Calgary, 3330 Hospital Dr. N.W., Calgary, Alberta, T2N 4N1, Canada

    Collin Luk & Naweed Syed

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  1. Dolores Martinez
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  2. Christophe Py
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  3. Mike W. Denhoff
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  4. Marzia Martina
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Correspondence to Dolores Martinez.

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Martinez, D., Py, C., Denhoff, M.W. et al. High-fidelity patch-clamp recordings from neurons cultured on a polymer microchip. Biomed Microdevices 12, 977–985 (2010). https://doi.org/10.1007/s10544-010-9452-z

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