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Title: Hydrodynamic trapping for rapid assembly and in situ electrical characterization of droplet interface bilayer arrays

Abstract

The droplet interface bilayer (DIB) is a modular technique for assembling planar lipid membranes between water droplets in oil. The DIB method thus provides a unique capability for developing digital, droplet-based membrane platforms for rapid membrane characterization, drug screening and ion channel recordings. This paper demonstrates a new, low-volume microfluidic system that automates droplet generation, sorting, and sequential trapping in designated locations to enable the rapid assembly of arrays of DIBs. The channel layout of the device is guided by an equivalent circuit model, which predicts that a serial arrangement of hydrodynamic DIB traps enables sequential droplet placement and minimizes the hydrodynamic pressure developed across filled traps to prevent squeeze-through of trapped droplets. Furthermore, the incorporation of thin-film electrodes fabricated via evaporation metal deposition onto the glass substrate beneath the channels allows for the first time in situ, simultaneous electrical interrogation of multiple DIBs within a sealed device. Combining electrical measurements with imaging enables measurements of membrane capacitance and resistance and bilayer area, and our data show that DIBs formed in different trap locations within the device exhibit similar sizes and transport properties. Simultaneous, single channel recordings of ion channel gating in multiple membranes are obtained when alamethicin peptides aremore » incorporated into the captured droplets, qualifying the thin-film electrodes as a means for measuring stimuli-responsive functions of membrane-bound biomolecules. Furthermore, this novel microfluidic-electrophysiology platform provides a reproducible, high throughput method for performing electrical measurements to study transmembrane proteins and biomembranes in low-volume, droplet-based membranes.« less

Authors:
 [1];  [2];  [2];  [3];  [1]
+ Show Author Affiliations
  1. Univ. of Tennessee, Knoxville, TN (United States)
  2. Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
  3. UT-Battelle, Oak Ridge, TN (United States)
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Nanophase Materials Sciences (CNMS)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1334470
Grant/Contract Number:  
AC05-00OR22725
Resource Type:
Accepted Manuscript
Journal Name:
Lab on a Chip
Additional Journal Information:
Journal Volume: 16; Journal Issue: 18; Journal ID: ISSN 1473-0197
Publisher:
Royal Society of Chemistry
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY

Citation Formats

Nguyen, Mary -Anne, Srijanto, Bernadeta, Collier, C. Patrick, Retterer, Scott T., and Sarles, Stephen A. Hydrodynamic trapping for rapid assembly and in situ electrical characterization of droplet interface bilayer arrays. United States: N. p., 2016. Web. doi:10.1039/C6LC00810K.
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Nguyen, Mary -Anne, Srijanto, Bernadeta, Collier, C. Patrick, Retterer, Scott T., & Sarles, Stephen A. Hydrodynamic trapping for rapid assembly and in situ electrical characterization of droplet interface bilayer arrays. United States. https://doi.org/10.1039/C6LC00810K
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Nguyen, Mary -Anne, Srijanto, Bernadeta, Collier, C. Patrick, Retterer, Scott T., and Sarles, Stephen A. Tue . "Hydrodynamic trapping for rapid assembly and in situ electrical characterization of droplet interface bilayer arrays". United States. https://doi.org/10.1039/C6LC00810K. https://www.osti.gov/servlets/purl/1334470.
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@article{osti_1334470,
title = {Hydrodynamic trapping for rapid assembly and in situ electrical characterization of droplet interface bilayer arrays},
author = {Nguyen, Mary -Anne and Srijanto, Bernadeta and Collier, C. Patrick and Retterer, Scott T. and Sarles, Stephen A.},
abstractNote = {The droplet interface bilayer (DIB) is a modular technique for assembling planar lipid membranes between water droplets in oil. The DIB method thus provides a unique capability for developing digital, droplet-based membrane platforms for rapid membrane characterization, drug screening and ion channel recordings. This paper demonstrates a new, low-volume microfluidic system that automates droplet generation, sorting, and sequential trapping in designated locations to enable the rapid assembly of arrays of DIBs. The channel layout of the device is guided by an equivalent circuit model, which predicts that a serial arrangement of hydrodynamic DIB traps enables sequential droplet placement and minimizes the hydrodynamic pressure developed across filled traps to prevent squeeze-through of trapped droplets. Furthermore, the incorporation of thin-film electrodes fabricated via evaporation metal deposition onto the glass substrate beneath the channels allows for the first time in situ, simultaneous electrical interrogation of multiple DIBs within a sealed device. Combining electrical measurements with imaging enables measurements of membrane capacitance and resistance and bilayer area, and our data show that DIBs formed in different trap locations within the device exhibit similar sizes and transport properties. Simultaneous, single channel recordings of ion channel gating in multiple membranes are obtained when alamethicin peptides are incorporated into the captured droplets, qualifying the thin-film electrodes as a means for measuring stimuli-responsive functions of membrane-bound biomolecules. Furthermore, this novel microfluidic-electrophysiology platform provides a reproducible, high throughput method for performing electrical measurements to study transmembrane proteins and biomembranes in low-volume, droplet-based membranes.},
doi = {10.1039/C6LC00810K},
journal = {Lab on a Chip},
number = 18,
volume = 16,
place = {United States},
year = {Tue Aug 02 00:00:00 EDT 2016},
month = {Tue Aug 02 00:00:00 EDT 2016}
}
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Publisher's Version of Record
https://doi.org/10.1039/C6LC00810K
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    • Challita, Elio J.; Makhoul-Mansour, Michelle M.; Freeman, Eric C.
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