Constructing droplet interface bilayers from the contact of aqueous droplets in oil.
We describe a protocol for forming an artificial lipid bilayer by contacting nanoliter aqueous droplets in an oil solution in the presence of phospholipids. A lipid monolayer forms at each oil-water interface, and when two such monolayers touch, a bilayer is created. Droplet interface bilayers (DIBs...
| Main Authors: | , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
2013
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| _version_ | 1826282861906362368 |
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| author | Leptihn, S Castell, O Cronin, B Lee, E Gross, L Marshall, D Thompson, JR Holden, M Wallace, M |
| author_facet | Leptihn, S Castell, O Cronin, B Lee, E Gross, L Marshall, D Thompson, JR Holden, M Wallace, M |
| author_sort | Leptihn, S |
| collection | OXFORD |
| description | We describe a protocol for forming an artificial lipid bilayer by contacting nanoliter aqueous droplets in an oil solution in the presence of phospholipids. A lipid monolayer forms at each oil-water interface, and when two such monolayers touch, a bilayer is created. Droplet interface bilayers (DIBs) are a simple way to generate stable bilayers suitable for single-channel electrophysiology and optical imaging from a wide variety of preparations, ranging from purified proteins to reconstituted eukaryotic cell membrane fragments. Examples include purified proteins from the α-hemolysin pore from Staphylococcus aureus, the anthrax toxin pore and the 1.2-MDa mouse mechanosensitive channel MmPiezo1. Ion channels and ionotropic receptors can also be reconstituted from membrane fragments without further purification. We describe two approaches for forming DIBs. In one approach, a lipid bilayer is created between two aqueous droplets submerged in oil. In the other approach, a membrane is formed between an aqueous droplet and an agarose hydrogel, which allows imaging in addition to electrical recordings. The protocol takes <30 min, including droplet generation, monolayer assembly and bilayer formation. In addition to the main protocol, we also describe the preparation of Ag/AgCl electrodes and sample preparation. |
| first_indexed | 2024-03-07T00:50:14Z |
| format | Journal article |
| id | oxford-uuid:86218d23-3c29-4a52-b81c-97e9886a673d |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T00:50:14Z |
| publishDate | 2013 |
| record_format | dspace |
| spelling | oxford-uuid:86218d23-3c29-4a52-b81c-97e9886a673d2022-03-26T22:02:04ZConstructing droplet interface bilayers from the contact of aqueous droplets in oil.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:86218d23-3c29-4a52-b81c-97e9886a673dEnglishSymplectic Elements at Oxford2013Leptihn, SCastell, OCronin, BLee, EGross, LMarshall, DThompson, JRHolden, MWallace, MWe describe a protocol for forming an artificial lipid bilayer by contacting nanoliter aqueous droplets in an oil solution in the presence of phospholipids. A lipid monolayer forms at each oil-water interface, and when two such monolayers touch, a bilayer is created. Droplet interface bilayers (DIBs) are a simple way to generate stable bilayers suitable for single-channel electrophysiology and optical imaging from a wide variety of preparations, ranging from purified proteins to reconstituted eukaryotic cell membrane fragments. Examples include purified proteins from the α-hemolysin pore from Staphylococcus aureus, the anthrax toxin pore and the 1.2-MDa mouse mechanosensitive channel MmPiezo1. Ion channels and ionotropic receptors can also be reconstituted from membrane fragments without further purification. We describe two approaches for forming DIBs. In one approach, a lipid bilayer is created between two aqueous droplets submerged in oil. In the other approach, a membrane is formed between an aqueous droplet and an agarose hydrogel, which allows imaging in addition to electrical recordings. The protocol takes <30 min, including droplet generation, monolayer assembly and bilayer formation. In addition to the main protocol, we also describe the preparation of Ag/AgCl electrodes and sample preparation. |
| spellingShingle | Leptihn, S Castell, O Cronin, B Lee, E Gross, L Marshall, D Thompson, JR Holden, M Wallace, M Constructing droplet interface bilayers from the contact of aqueous droplets in oil. |
| title | Constructing droplet interface bilayers from the contact of aqueous droplets in oil. |
| title_full | Constructing droplet interface bilayers from the contact of aqueous droplets in oil. |
| title_fullStr | Constructing droplet interface bilayers from the contact of aqueous droplets in oil. |
| title_full_unstemmed | Constructing droplet interface bilayers from the contact of aqueous droplets in oil. |
| title_short | Constructing droplet interface bilayers from the contact of aqueous droplets in oil. |
| title_sort | constructing droplet interface bilayers from the contact of aqueous droplets in oil |
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