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Rapid Microfluidic Perfusion Enabling Kinetic Studies of Lipid Ion Channels in a Bilayer Lipid Membrane Chip

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Abstract

There is growing recognition that lipids play key roles in ion channel physiology, both through the dynamic formation and dissolution of lipid ion channels and by indirect regulation of protein ion channels. Because existing technologies cannot rapidly modulate the local (bio)chemical conditions at artificial bilayer lipid membranes used in ion channel studies, the ability to elucidate the dynamics of these lipid–lipid and lipid–protein interactions has been limited. Here we demonstrate a microfluidic system supporting exceptionally rapid perfusion of reagents to an on-chip bilayer lipid membrane, enabling the responses of lipid ion channels to dynamic changes in membrane boundary conditions to be probed. The thermoplastic microfluidic system allows initial perfusion of reagents to the membrane in less than 1 s, and enables kinetic behaviors with time constants below 10 s to be directly measured. Application of the platform is demonstrated toward kinetic studies of ceramide, a biologically important lipid known to self-assemble into transmembrane ion channels, in response to dynamic treatments of small ions (La3+) and proteins (Bcl-xL mutant). The results reveal the broader potential of the technology for studies of membrane biophysics, including lipid ion channel dynamics, lipid–protein interactions, and the regulation of protein ion channels by lipid micro domains.

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Acknowledgments

The authors gratefully acknowledge Dr. Louis Hromada for input on chip fabrication, Meenu Perera for assistance with pClamp software and Vidyaramanan Ganesan for providing the Bcl-xL mutant. We thank Dr. David W. Andrews at McMaster University in Canada for providing plasmid for protein preparation. This work is supported by NIH grants R01GM072512 and R21EB011750.

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Authors and Affiliations

  1. Department of Mechanical Engineering, University of Maryland, 3123 Glenn L Martin Hall, College Park, MD, 20742, USA

    Chenren Shao

  2. Department of Biology, University of Maryland, College Park, MD, USA

    Bing Sun & Marco Colombini

  3. Department of Mechanical Engineering, University of Maryland, 3139 Glenn L Martin Hall, College Park, MD, 20742, USA

    Don L. DeVoe

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  1. Chenren Shao
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  2. Bing Sun
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  3. Marco Colombini
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  4. Don L. DeVoe
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Correspondence to Don L. DeVoe.

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Associate Editor Tingrui Pan oversaw the review of this article.

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Shao, C., Sun, B., Colombini, M. et al. Rapid Microfluidic Perfusion Enabling Kinetic Studies of Lipid Ion Channels in a Bilayer Lipid Membrane Chip. Ann Biomed Eng 39, 2242–2251 (2011). https://doi.org/10.1007/s10439-011-0323-4

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