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Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses

We present a chip design allowing rapid and robust lipid bilayer (LBL) membrane formation using a Parylene coated thin silicon nitride aperture. After bilayer formation, single membrane channels can be reconstituted and characterized by electrophysiology. The ability for robust reconstitution will a...

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Autores principales: Ahmed, Tanzir, van den Driesche, Sander, Bafna, Jayesh Arun, Oellers, Martin, Hemmler, Roland, Gall, Karsten, Wagner, Richard, Winterhalter, Mathias, Vellekoop, Michael J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192868/
https://www.ncbi.nlm.nih.gov/pubmed/32355998
http://dx.doi.org/10.1007/s10544-020-0473-y
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author Ahmed, Tanzir
van den Driesche, Sander
Bafna, Jayesh Arun
Oellers, Martin
Hemmler, Roland
Gall, Karsten
Wagner, Richard
Winterhalter, Mathias
Vellekoop, Michael J.
author_facet Ahmed, Tanzir
van den Driesche, Sander
Bafna, Jayesh Arun
Oellers, Martin
Hemmler, Roland
Gall, Karsten
Wagner, Richard
Winterhalter, Mathias
Vellekoop, Michael J.
author_sort Ahmed, Tanzir
collection PubMed
description We present a chip design allowing rapid and robust lipid bilayer (LBL) membrane formation using a Parylene coated thin silicon nitride aperture. After bilayer formation, single membrane channels can be reconstituted and characterized by electrophysiology. The ability for robust reconstitution will allow parallelization and enhanced screening of small molecule drugs acting on or permeating across the membrane channel. The aperture was realized on a microfabricated silicon nitride membrane by using standard clean-room fabrication processes. To ensure the lipid bilayer formation, the nitride membrane was coated with a hydrophobic and biocompatible Parylene layer. We tested both Parylene-C and Parylene-AF4. The contact angle measurements on both Parylene types showed very good hydrophobic properties and affinity to lipids. No precoating of the Parylene with an organic solvent is needed to make the aperture lipophilic, in contradiction to Teflon membranes. The chips can be easily placed in an array utilizing a 3D printed platform. Experiments show repetitive LBL formation and destruction (more than 6 times) within a very short time (few seconds). Through measurements we have established that the LBL layers are very thin. This allows the investigation of the fusion process of membrane proteins i.e. outer membrane protein (OmpF) in the LBL within a few minutes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10544-020-0473-y) contains supplementary material, which is available to authorized users.
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spelling pubmed-71928682020-05-05 Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses Ahmed, Tanzir van den Driesche, Sander Bafna, Jayesh Arun Oellers, Martin Hemmler, Roland Gall, Karsten Wagner, Richard Winterhalter, Mathias Vellekoop, Michael J. Biomed Microdevices Article We present a chip design allowing rapid and robust lipid bilayer (LBL) membrane formation using a Parylene coated thin silicon nitride aperture. After bilayer formation, single membrane channels can be reconstituted and characterized by electrophysiology. The ability for robust reconstitution will allow parallelization and enhanced screening of small molecule drugs acting on or permeating across the membrane channel. The aperture was realized on a microfabricated silicon nitride membrane by using standard clean-room fabrication processes. To ensure the lipid bilayer formation, the nitride membrane was coated with a hydrophobic and biocompatible Parylene layer. We tested both Parylene-C and Parylene-AF4. The contact angle measurements on both Parylene types showed very good hydrophobic properties and affinity to lipids. No precoating of the Parylene with an organic solvent is needed to make the aperture lipophilic, in contradiction to Teflon membranes. The chips can be easily placed in an array utilizing a 3D printed platform. Experiments show repetitive LBL formation and destruction (more than 6 times) within a very short time (few seconds). Through measurements we have established that the LBL layers are very thin. This allows the investigation of the fusion process of membrane proteins i.e. outer membrane protein (OmpF) in the LBL within a few minutes. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s10544-020-0473-y) contains supplementary material, which is available to authorized users. Springer US 2020-04-30 2020 /pmc/articles/PMC7192868/ /pubmed/32355998 http://dx.doi.org/10.1007/s10544-020-0473-y Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Ahmed, Tanzir
van den Driesche, Sander
Bafna, Jayesh Arun
Oellers, Martin
Hemmler, Roland
Gall, Karsten
Wagner, Richard
Winterhalter, Mathias
Vellekoop, Michael J.
Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses
title Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses
title_full Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses
title_fullStr Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses
title_full_unstemmed Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses
title_short Rapid lipid bilayer membrane formation on Parylene coated apertures to perform ion channel analyses
title_sort rapid lipid bilayer membrane formation on parylene coated apertures to perform ion channel analyses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192868/
https://www.ncbi.nlm.nih.gov/pubmed/32355998
http://dx.doi.org/10.1007/s10544-020-0473-y
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