Flow cell for chemical reactions

Abstract

Devices and methods are disclosed for synthesizing compounds on the surface of supports. The devices are flow devices, which include a housing comprising a housing chamber. The housing has an opening adapted for insertion of a support into the housing chamber. A sealing member is movably mounted in the housing chamber and adapted to engage the support to form a reagent chamber between a surface of the support and a surface of the sealing member. A mechanism is included for moving the sealing member within the housing chamber. The device has both an inlet and an outlet, which are both in fluid communication with the reagent chamber. In the methods of the invention a support is placed into a chamber of a device such as described above. The mechanism adapted to engage the support on a surface opposite the surface engaged by the sealing member is activated to urge the support toward the sealing member. The pressure-activated mechanism is activated to urge the support against the aforesaid mechanism and against an interior wall of the housing chamber to form the reagent chamber. A fluid reagent for conducting the reaction step is introduced into the reagent chamber by means of the inlet. Thereafter, the fluid reagent is removed from the reagent chamber. The pressure-activated mechanism is deactivated and the support is removed from the housing chamber. In this way the reagent chamber is formed and un-formed in situ.

What is claimed is:  
     
         1 . A flow device comprising: 
 (a) a housing comprising a housing chamber,    (b) an opening in said housing adapted for insertion of a support into said housing chamber,    (c) a sealing member movably mounted in said housing chamber and adapted to engage said support to form a reagent chamber between a surface of said support and a surface of said sealing member,    (d) a mechanism for moving said sealing member within said housing chamber,    (e) an inlet in fluid communication with said reagent chamber and    (f) an outlet in fluid communication with said reagent chamber.    
     
     
         2 . A device according to  claim 1  further comprising a viewing area in a wall of said housing adapted to view a surface of said support.  
     
     
         3 . A device according to  claim 1  wherein said viewing area is a window in a wall of said housing.  
     
     
         4 . A device according to  claim 3  further comprising a mechanism adapted to engage said support on a surface opposite said surface engaged by said sealing member.  
     
     
         5 . A device according to  claim 4  wherein said mechanism adapted to engage said support comprises two to four pressure-activated cylinders, each of which is disposed in a port in said window.  
     
     
         6 . A device according to  claim 1  wherein said support is engaged by a holding element and said opening in said housing is adapted to receive said support secured by said holding element.  
     
     
         7 . A device according to  claim 6  wherein said holding element is a vacuum actuated element.  
     
     
         8 . A device according to  claim 6  wherein said holding element is part of a robotic arm.  
     
     
         9 . A device according to  claim 1  further comprising a pressure-activated mechanism to which said sealing member is attached.  
     
     
         10 . A device according to  claim 9  wherein said pressure-activated mechanism is a pressure-activated cylinder.  
     
     
         11 . A device according to  claim 10  wherein said cylinder further comprises a guide.  
     
     
         12 . A flow device comprising: 
 (a) a housing comprising a housing chamber,    (b) an opening in said housing adapted for insertion of a holding element comprising a support into said housing chamber,    (c) a sealing member movably mounted in said housing chamber and adapted to engage said support to form a reagent chamber between a surface of said support and a surface of said sealing member, wherein said sealing member is attached to a pressure-activated mechanism for moving said sealing member within said housing chamber,    (d) a mechanism adapted to engage said support on a surface opposite said surface engaged by said sealing member,    (e) an inlet in fluid communication with said reagent chamber,    (f) an outlet in fluid communication with said reagent chamber, and    (g) a viewing area in a wall of said housing adapted to view a surface of said support.    
     
     
         13 . A device according to  claim 12  wherein said viewing area is a window in a wall of said housing.  
     
     
         14 . A device according to  claim 12  wherein said mechanism adapted to engage said support on a surface opposite said surface engaged by said sealing member comprises two to four pressure-activated cylinders, each of which is disposed in a port in said window.  
     
     
         15 . A device according to  claim 14  wherein said holding element is a vacuum actuated element.  
     
     
         16 . A device according to  claim 15  wherein said holding element is part of a robotic arm.  
     
     
         17 . A device according to  claim 12  said pressure-activated mechanism is a pressure-activated cylinder within a guide.  
     
     
         18 . A device according to  claim 12  wherein said device is rotatably mounted on a frame.  
     
     
         19 . A method for performing a step of a chemical reaction on the surface of a support, said method comprising: 
 (a) placing a support into a chamber of a device, said device comprising: 
 (i) a housing comprising a housing chamber,  
 (ii) an opening in said housing adapted for insertion of a holding element comprising a support into said housing chamber,  
 (iii) a sealing member movably mounted in said housing chamber and adapted to engage said support to form a reagent chamber between a surface of said support and a surface of said sealing member, wherein said sealing member is attached to a pressure-activated mechanism for moving said sealing member within said housing chamber,  
 (iv) a mechanism adapted to engage said support on a surface opposite said surface engaged by said sealing member,  
 (v) an inlet in fluid communication with said reagent chamber,  
 (vi) an outlet in fluid communication with said reagent chamber, and  
 (vii) a viewing area in a wall of said housing adapted to view a surface of said support;  
   (b) activating said mechanism of (iv) to urge said support toward said sealing member,    (c) activating said pressure-activated mechanism of (iii) to urge said support against said mechanism of (iv) and against an interior wall of said housing chamber and to thereby form said reagent chamber,    (d) introducing a fluid reagent for conducting said reaction step into said reagent chamber by means of said inlet,    (e) removing said fluid reagent from said reagent chamber by means of said outlet,    (f) deactivating said pressure-activated mechanism, and    (g) removing said support from said housing chamber.    
     
     
         20 . A method according to  claim 19  wherein said chemical reaction is selected from the group consisting of washing said surface, oxidizing a substance attached to said surface, removing a protecting group from a substance on said surface, blocking and deblocking sites on said surface and reducing a substance attached to said surface.  
     
     
         21 . A method according to  claim 19  wherein said viewing area is a window in a wall of said housing and said method further comprises viewing said surface of said support to determine the status of said step of said chemical reaction.  
     
     
         22 . A method according to  claim 21  wherein said mechanism of (iv) comprises two to four pressure-activated cylinders, each of which is disposed in a port in said window.  
     
     
         23 . A method according to  claim 19  wherein said holding element is a vacuum actuated element and said method further comprises actuating said element to thereby secure said support to said element.  
     
     
         24 . A method according to  claim 19  wherein said pressure-activated mechanism of (iii) is a pressure-activated cylinder within a guide.  
     
     
         25 . A method according to  claim 19  wherein said device is rotatably mounted on a frame and said method comprises rotating said device 45 degrees after introduction of said fluid reagent in step (d).  
     
     
         26 . A method for performing a step of a chemical reaction on the surface of a support, said method comprising: 
 (a) placing a support into a chamber of a device,    (b) activating a mechanism adapted to engage said support on a surface thereof,    (c) activating a pressure-activated mechanism for moving a sealing member within said chamber to engage a surface of said support opposite said surface engaged by said mechanism of step (b) and deactivating said mechanism of step (b) to thereby form a reagent chamber,    (d) introducing a fluid reagent for conducting said reaction step into said reagent chamber,    (e) removing said fluid reagent from said reagent chamber,    (f) deactivating said pressure-activated mechanism, and    (g) removing said support from said chamber.    
     
     
         27 . A method according to  claim 26  wherein said chemical reaction is selected from the group consisting of washing said surface, oxidizing a substance attached to said surface, removing a protecting group from a substance on said surface, blocking and deblocking sites on said surface and reducing a substance attached to said surface.  
     
     
         28 . A method according to  claim 26  wherein said device is rotatably mounted on a frame and said method comprises rotating said device about 20 to about 45 degrees after introduction of said fluid reagent in step (d).  
     
     
         29 . A method for synthesizing a plurality of biopolymers on the surface of a support wherein said synthesis comprises a plurality of monomer additions, said method comprising after each of said monomer additions: 
 (a) placing said support into a chamber of a first device according to  claim 1  and subjecting said surface to a first step of said synthesis that is subsequent to a monomer addition and    (b) placing said support into a chamber of a second device according to  claim 1  and subjecting said surface to a second step of said synthesis that is subsequent to said first step.    
     
     
         30 . A method according to  claim 29  wherein each of said first step and said second step comprises a wash.  
     
     
         31 . A method according to  claim 29  wherein said biopolymers are polynucleotides.  
     
     
         32 . A method according to  claim 31  wherein said first step comprises subjecting said surface to an oxidizing agent.  
     
     
         33 . A method according to  claim 31  wherein said second step comprises subjecting said surface to an agent for removing a protecting group.  
     
     
         34 . A method according to  claim 29  wherein a wash solution and a reagent for said synthesis are independently directed to said inlet of said device.  
     
     
         35 . A method according to  claim 34  wherein said wash solution is an organic solvent.  
     
     
         36 . A method according to  claim 35  wherein said biopolymers are synthesized on said surface in an array.  
     
     
         37 . A method according to  claim 36  wherein said biopolymers are synthesized on said surface in multiple arrays and said support is subsequently diced into individual arrays of biopolymers on a support.  
     
     
         38 . An apparatus for synthesizing an array of biopolymers on the surface of a support, said apparatus comprising: 
 (a) a plurality of devices according to  claim 1  rotatably mounted on said apparatus,    (b) one or more fluid dispensing stations in fluid communication with one or more of said devices,    (c) a station for monomer addition to said surface of said support,    (d) a mechanism, comprising said holding element, for moving a support to and from said station for monomer addition and one of said devices and from one of said devices to another of said devices, and    (e) a mechanism for rotating said devices.    
     
     
         39 . An apparatus according to  claim 38  further comprising a controller for controlling the movement of said mechanism for moving said support.  
     
     
         40 . An apparatus according to  claim 38  wherein said mechanism is a robotic arm.  
     
     
         41 . An apparatus according to  claim 38  wherein said apparatus further comprises a manifold in fluid communication with an inlet of one of said devices.  
     
     
         42 . A flow device comprising: 
 (a) a sealing member adapted for sealing engagement with a support to form a reagent chamber in situ,    (b) an inlet in fluid communication with said reagent chamber, and    (c) an outlet in fluid communication with said reagent chamber.    
     
     
         43 . A method for performing a step of a chemical reaction on the surface of a support, said method comprising: 
 (a) forming a reaction chamber in situ, said reaction chamber comprising said surface of said support and a sealing member,    (b) introducing a fluid reagent for conducting said reaction step into said reagent chamber,    (c) removing said fluid reagent from said reagent chamber, and    (d) removing said support from contact with said sealing member to thereby un-form said reaction chamber.    
     
     
         44 . A flow device comprising: 
 (a) a housing comprising a housing chamber,    (b) an opening in said housing adapted for insertion of a support into said housing chamber,    (c) a sealing member movably mounted in said housing chamber and adapted to engage said support to form a reagent chamber between a surface of said support and a surface of said sealing member,    (d) a mechanism for moving said sealing member within said housing chamber,    (e) an inlet in fluid communication with said reagent chamber,    (f) an outlet in fluid communication with said reagent chamber, and    (g) a viewing area in a wall of said housing adapted to view a surface of said support.    
     
     
         44 . A device according to  claim 43  wherein said viewing area is a window in a wall of said housing.  
     
     
         45 . A device for transferring a support from one flow cell to another flow cell, said device comprising: 
 (a) a vacuum actuated element for holding said support and    (b) a mechanism for moving the vacuum actuated element from one flow cell to another flow.    
     
     
         46 . A device according to  claim 45  wherein the vacuum actuated element comprises at least two prongs.  
     
     
         47 . A device according to  claim 45  wherein the mechanism is part of a robotic arm.  
     
     
         48 . A method for synthesizing an array of biopolymers on a surface of a support, said method comprising: 
 (a) adding one or more polymer subunits at each of multiple feature locations during each of multiple rounds of subunit additions and    (b) between rounds of subunit additions: 
 (i) forming a reaction chamber comprising said surface of said support and a member which seals against said surface,  
 (ii) introducing a fluid reagent for conducting said reaction step into said reagent chamber, and  
 (iii) removing said fluid reagent from said reagent chamber.  
   
     
     
         49 . A method according to  claim 48  additionally comprising removing said support from contact with said sealing member to thereby un-form said reaction chamber.