title: Constructing droplet interface bilayers from the contact of aqueous droplets in oil.
abstract: 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.
authors: Department of Chemistry, Oxford University, Oxford, UK.
authors: Sebastian
authors: S
authors: Leptihn
authors: Oliver K
authors: OK
authors: Castell
authors: Brid
authors: B
authors: Cronin
authors: En-Hsin
authors: EH
authors: Lee
authors: Linda C M
authors: LC
authors: Gross
authors: David P
authors: DP
authors: Marshall
authors: James R
authors: JR
authors: Thompson
authors: Matthew
authors: M
authors: Holden
authors: Mark I
authors: MI
authors: Wallace
mesh_terms: D000473:Alkanes; D000818:Animals; D000942:Antigens, Bacterial / Q000737:chemistry; D001427:Bacterial Toxins / Q000737:chemistry; D006460:Hemolysin Proteins / Q000737:chemistry; D007473:Ion Channels / Q000737:chemistry / Q000378:metabolism; D008051:Lipid Bilayers / Q000138:chemical synthesis* / Q000737:chemistry; D008567:Membranes, Artificial; D051379:Mice; D049329:Nanostructures / Q000737:chemistry*; D009821:Oils / Q000737:chemistry*; D012834:Silver; D018030:Silver Compounds; D014867:Water / Q000737:chemistry*
chemical_list: D000473:Alkanes; D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D009821:Oils; C552920:Piezo1 protein, mouse; D018030:Silver Compounds; C030325:anthrax toxin; C021091:staphylococcal alpha-toxin; D014867:Water; D012834:Silver; C007932:n-hexadecane; C016486:heptadecane; C037548:silver chloride
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