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Shaped apertures in photoresist films enhance the lifetime and mechanical stability of suspended lipid bilayers

Shaped apertures in photoresist films enhance the lifetime and mechanical stability of suspended lipid bilayers
Shaped apertures in photoresist films enhance the lifetime and mechanical stability of suspended lipid bilayers
Planar lipid bilayers suspended in apertures provide a controlled environment for ion channel studies. However, short lifetimes and poor mechanical stability of suspended bilayers limit the experimental throughput of bilayer electrophysiology experiments. Although bilayers are more stable in smaller apertures, ion channel incorporation through vesicle fusion with the suspended bilayer becomes increasingly difficult. In an alternative bilayer stabilization approach, we have developed shaped apertures in SU8 photoresist that have tapered sidewalls and a minimum diameter between 60 and 100 µm. Bilayers formed at the thin tip of these shaped apertures, either with the painting or the folding method, display drastically increased lifetimes, typically >20 hours, and mechanical stability, being able to withstand extensive perturbation of the buffer solution. Single-channel electrical recordings of the peptide alamethicin and of the proteoliposome-delivered potassium channel KcsA demonstrate channel conductance with low noise, made possible by the small capacitance of the 50 µm thick SU8 septum, which is only thinned around the aperture, and unimpeded proteoliposome fusion, enabled by the large aperture diameter. We anticipate that these shaped apertures with micrometer edge thickness can substantially enhance the throughput of channel characterisation by bilayer lipid membrane electrophysiology, especially in combination with automated parallel bilayer platforms.
0006-3495
1650-1659
Kalsi, Sumit
65cb3d4c-c28c-490a-85a3-7a8fb96ecfee
Powl, Andrew M.
79b77ec3-767b-42ad-991c-d843e29af8e3
Wallace, B.A.
ec92996f-4630-4e2a-a9b0-852fed8cdd9d
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
de Planque, Maurits R.R.
a1d33d13-f516-44fb-8d2c-c51d18bc21ba
Kalsi, Sumit
65cb3d4c-c28c-490a-85a3-7a8fb96ecfee
Powl, Andrew M.
79b77ec3-767b-42ad-991c-d843e29af8e3
Wallace, B.A.
ec92996f-4630-4e2a-a9b0-852fed8cdd9d
Morgan, Hywel
de00d59f-a5a2-48c4-a99a-1d5dd7854174
de Planque, Maurits R.R.
a1d33d13-f516-44fb-8d2c-c51d18bc21ba

Kalsi, Sumit, Powl, Andrew M., Wallace, B.A., Morgan, Hywel and de Planque, Maurits R.R. (2014) Shaped apertures in photoresist films enhance the lifetime and mechanical stability of suspended lipid bilayers. Biophysical Journal, 106 (8), 1650-1659. (doi:10.1016/j.bpj.2014.02.033).

Record type: Article

Abstract

Planar lipid bilayers suspended in apertures provide a controlled environment for ion channel studies. However, short lifetimes and poor mechanical stability of suspended bilayers limit the experimental throughput of bilayer electrophysiology experiments. Although bilayers are more stable in smaller apertures, ion channel incorporation through vesicle fusion with the suspended bilayer becomes increasingly difficult. In an alternative bilayer stabilization approach, we have developed shaped apertures in SU8 photoresist that have tapered sidewalls and a minimum diameter between 60 and 100 µm. Bilayers formed at the thin tip of these shaped apertures, either with the painting or the folding method, display drastically increased lifetimes, typically >20 hours, and mechanical stability, being able to withstand extensive perturbation of the buffer solution. Single-channel electrical recordings of the peptide alamethicin and of the proteoliposome-delivered potassium channel KcsA demonstrate channel conductance with low noise, made possible by the small capacitance of the 50 µm thick SU8 septum, which is only thinned around the aperture, and unimpeded proteoliposome fusion, enabled by the large aperture diameter. We anticipate that these shaped apertures with micrometer edge thickness can substantially enhance the throughput of channel characterisation by bilayer lipid membrane electrophysiology, especially in combination with automated parallel bilayer platforms.

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More information

Accepted/In Press date: 26 February 2014
e-pub ahead of print date: 15 April 2014
Published date: 15 April 2014
Organisations: Nanoelectronics and Nanotechnology

Identifiers

Local EPrints ID: 359987
URI: https://eprints.soton.ac.uk/id/eprint/359987
ISSN: 0006-3495
PURE UUID: 5309fe7b-d54e-4540-abba-f5269614539a
ORCID for Hywel Morgan: ORCID iD orcid.org/0000-0003-4850-5676
ORCID for Maurits R.R. de Planque: ORCID iD orcid.org/0000-0002-8787-0513

Catalogue record

Date deposited: 19 Nov 2013 15:38
Last modified: 20 Jul 2019 00:59

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