Formation of Layers of Amphiphilic Molecules

Abstract

To form a layer separating two volumes of aqueous solution, there is used an apparatus comprising elements defining a chamber, the elements including a body of non-conductive material having formed therein at least one recess opening into the chamber, the recess containing an electrode. A pre-treatment coating of a hydrophobic fluid is applied to the body across the recess. Aqueous solution, having amphiphilic molecules added thereto, is flowed across the body to cover the recess so that aqueous solution is introduced into the recess from the chamber and a layer of the amphiphilic molecules forms across the recess separating a volume of aqueous solution introduced into the recess from the remaining volume of aqueous solution.

1 . A method of forming a layer separating two volumes of aqueous solution, the method comprising:
 (a) providing an apparatus comprising elements defining a chamber, the elements including a body of non-conductive material having formed therein at least one recess opening into the chamber, the recess containing an electrode;   (b) applying a pre-treatment coating of a hydrophobic fluid to the body across the recess;   (c) flowing aqueous solution, having amphiphilic molecules added thereto, across the body to cover the recess so that aqueous solution is introduced into the recess from the chamber and a layer of the amphiphilic molecules forms across the recess separating a volume of aqueous solution introduced into the recess from the remaining volume of aqueous solution.   
     
     
         2 . A method according to  claim 1 , wherein step (c) comprises:
 (c1) flowing aqueous solution across the body to cover the recess so that aqueous solution flows into the recess;   (c2) flowing the aqueous solution to uncover the recess, leaving some aqueous solution in the recess; and   (c3) flowing aqueous solution that is optionally the same aqueous solution as in step (c2), having amphiphilic molecules added thereto, across the body and to re-cover the recess so that a layer of the amphiphilic molecules forms across the recess separating a volume of aqueous solution inside the recess from the remaining volume of aqueous solution.   
     
     
         3 . A method according to  claim 2 , wherein
 the apparatus is provided with a further electrode in the chamber outside said recess, in step (c1), the aqueous solution is flowed also to contact the further electrode, and step (c) further comprises, between steps (c1) and (c2):   (c4) applying a voltage across said electrode contained in the recess and said further electrode sufficient to reduce the amount of excess hydrophobic fluid covering said electrode contained in the recess.   
     
     
         4 . (canceled) 
     
     
         5 . A method according to  claim 1 , wherein surfaces including one or both of (a) the outermost surface of the body around the recess, and (b) at least an outer part of the internal surface of the recess extending from the rim of the recess, are hydrophobic. 
     
     
         6 . A method according to  claim 5 , wherein the body comprises an outermost layer formed of a hydrophobic material, the recess extending through the outermost layer and said outer part of the internal surface of the recess being a surface of the outermost layer. 
     
     
         7 . A method according to  claim 5 , wherein an inner part of the internal surface of the recess inside the outer part is hydrophilic. 
     
     
         8 .- 11 . (canceled) 
     
     
         12 . A method according to  claim 1 , wherein the body comprises a substrate and at least one further layer attached to the substrate, the recess extending through the at least one further layer. 
     
     
         13 . A method according to any  claim 1 , wherein the electrode has provided thereon a hydrophillic surface which repels the hydrophobic fluid applied in step (c) whilst allowing ionic conduction from the aqueous solution to the electrode. 
     
     
         14 .- 18 . (canceled) 
     
     
         19 . A method according to  claim 1 , wherein the internal surface of the 15 recess has no openings capable of fluid communication. 
     
     
         20 .- 23 . (canceled) 
     
     
         24 . A method according to  claim 1 , further comprising, before step (c), depositing the amphiphilic molecules on an internal surface of the chamber or on an internal surface  30  in the flow path of the aqueous solution into the chamber, the aqueous solution covering the internal surface during step (c) whereby the amphiphilic molecules are added to the aqueous solution. 
     
     
         25 .- 27 . (canceled) 
     
     
         28 . A method according to  claim 1 , wherein the at least one recess comprises plural recesses and the method comprises inserting different membrane protein into the layers of amphiphilic molecules formed in different recesses. 
     
     
         29 . A method according to  claim 1 , further comprising inserting a membrane protein into the layer of amphiphilic molecules and wherein the apparatus is provided with a further electrode in the chamber outside the recess, and the method further comprises applying a potential across the electrode in the recess and the further electrode and monitoring an electrical signal developed between the electrode in the recess and the further electrode. 
     
     
         30 . An apparatus for supporting a layer separating two volumes of aqueous solution, the apparatus comprising:
 elements defining a chamber, the elements including a body of non-conductive material having formed therein at least one recess opening into the chamber; and   an electrode contained in the recess.   
     
     
         31 . An apparatus according to  claim 30 , wherein surfaces including either or both of (a) the outermost surface of the body around the recess, and (b) at least an outer part of the internal surface of the recess extending from the rim of the recess, are hydrophobic. 
     
     
         32 . An apparatus according to  claim 31 , wherein the body comprises an outermost layer formed of a hydrophobic material, the recess extending through the outermost layer and said outer part of the internal surface of the recess being a surface of the outermost layer. 
     
     
         33 . An apparatus according to  claim 31 , wherein an inner part of the internal surface of the recess inside the outer part is hydrophilic. 
     
     
         34 .- 37 . (canceled) 
     
     
         38 . An apparatus according to  claim 30 , wherein the body comprises a substrate and at least one further layer attached to the substrate, the recess extending through the at least one further layer. 
     
     
         39 .- 43 . (canceled) 
     
     
         44 . An apparatus according to  claim 30 , wherein the electrode has provided thereon a hydrophillic surface which repels the hydrophobic fluid applied in step (c) whilst allowing ionic conduction from the aqueous solution to the electrode. 
     
     
         45 .- 47 . (canceled) 
     
     
         48 . An apparatus according to  claim 30 , further comprising a further electrode in the chamber outside said recess. 
     
     
         49 . An apparatus according to  claim 30 , wherein the elements defining the chamber further include a cover extending over the body so that the chamber is a closed chamber, the cover comprising at least one inlet and at least one outlet. 
     
     
         50 . (canceled) 
     
     
         51 . An apparatus according to  claim 30 , wherein the internal surface of the recess has no openings capable of fluid communication. 
     
     
         52 .- 56 . (canceled) 
     
     
         57 . An apparatus according to  claim 30 , further comprising a pre-treatment coating of a hydrophobic fluid applied to the body across the recess. 
     
     
         58 .- 71 . (canceled)