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High Yield, Reproducible and Quasi-Automated Bilayer Formation in a Microfluidic Format

Authors

  • Verena C. Stimberg,

    1. BIOS–Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands
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  • Johan G. Bomer,

    1. BIOS–Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands
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  • Iris van Uitert,

    1. BIOS–Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands
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  • Albert van den Berg,

    1. BIOS–Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands
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  • Séverine Le Gac

    Corresponding author
    1. BIOS–Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands
    • BIOS–Lab on a Chip Group, MESA+ Institute for Nanotechnology, University of Twente, 7500 AE Enschede, The Netherlands.
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Abstract

A microfluidic platform is reported for various experimentation schemes on cell membrane models and membrane proteins using a combination of electrical and optical measurements, including confocal microscopy. Bilayer lipid membranes (BLMs) are prepared in the device upon spontaneous and instantaneous thinning of the lipid solution in a 100-μm dry-etched aperture in a 12.5-μm thick Teflon foil. Using this quasi-automated approach, a remarkable 100% membrane formation yield is reached (including reflushing in 4% of the cases), and BLMs are stable for up to 36 h. Furthermore, the potential of this platform is demonstrated for (i) the in-depth characterization of BLMs comprising both synthetic and natural lipids (1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and L-α-phosphatidylcholine (L-α-PC)/cholesterol, respectively) in terms of seal resistance, capacitance, surface area, specific capacitance, and membrane hydrophobic thickness; (ii) confocal microscopy imaging of phase separation in sphingomyelin/L-α-PC/cholesterol ternary membranes; (iii) electrical measurements of individual nanopores (α-hemolysin, gramicidin); and (iv) indirect assessment of the alteration of membrane properties upon exposure to chemical stimuli using the natural nanopore gramicidin as a sensor.

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