Microfluidic Devices and Methods of Use in The Formation and Control of Nanoreactors

Abstract

The present invention provides novel microfluidic devices and methods that are useful for performing high-throughput screening assays and combinatorial chemistry. Such methods can include labeling a library of compounds by emulsifying aqueous solutions of the compounds and aqueous solutions of unique liquid labels on a microfluidic device, which includes a plurality of electrically addressable, channel bearing fluidic modules integrally arranged on a microfabricated substrate such that a continuous channel is provided for flow of immiscible fluids, whereby each compound is labeled with a unique liquid label, pooling the labeled emulsions, coalescing the labeled emulsions with emulsions containing a specific cell or enzyme, thereby forming a nanoreactor, screening the nanoreactors for a desirable reaction between the contents of the nanoreactor, and decoding the liquid label, thereby identifying a single compound from a library of compounds.

1 - 29 . (canceled) 
     
     
         30 . A microfluidic device comprising a microfabricated substrate, said substrate further comprising:
 a) one or more inlet modules having at least one fluorinated inlet channel adapted to carry at least one dispersed phase fluid comprising an aqueous solution;   b) at least one fluorinated main channel adapted to carry at least one continuous phase fluid comprising a fluorinated oil, wherein said inlet channel is in fluid communication with said main channel such that said dispersed phase fluid is immiscible with said continuous phase fluid and forms a plurality of monodisperse droplets in said continuous phase fluid; and   c) a coalescence module downstream from and in fluid communication with said one or more inlet modules via the main channel, wherein two or more droplets passing there through are coalesced to form a nanoreactor, wherein the nanoreactor is about 25 μm in diameter.   
     
     
         31 . The device of  claim 30 , wherein the fluorinated oil further comprises a surfactant. 
     
     
         32 . The device of  claim 31 , wherein the surfactant is sorbitan-based carboxylic acid esters, perfluorinated polyethers, non-ionic surfactants or ionic surfactants. 
     
     
         33 . The device of  claim 30 , wherein said channels are coated with fluorosilane, an amorphous soluble perfluoropolymer, BSA, PEG-silane, a silica primer layer followed by a perfluoroalkylalkylsilane compound, Teflon or fluorinated oils. 
     
     
         34 . The device of  claim 33 , wherein the fluorinated oils are octadecafluoroctane, Fluorinert or fluorononane. 
     
     
         35 . The device of  claim 34 , wherein the fluorinated oil further comprises a surfactant. 
     
     
         36 . The device of  claim 30 , wherein said dispersed phase comprises a library of droplets. 
     
     
         37 . The device of  claim 36 , wherein said library of droplets comprises a biological/chemical material, wherein said biological/chemical material is selected from the group consisting of tissues, cells, particles, proteins, antibodies, amino acids, nucleotides, small molecules, and pharmaceuticals. 
     
     
         38 . The device of  claim 30 , wherein said inlet channel is further connected to a means for introducing a sample to said device. 
     
     
         39 . The device of  claim 30  further comprising a means for simultaneously loading multiple samples onto the device. 
     
     
         40 . The device of  claim 30 , further comprising one or more electrodes that generate an electric field. 
     
     
         41 . The device of  claim 40 , wherein the electrodes comprise metal alloy components and the metal alloy components are integrally contained in one or more channels and are isolated from the inlet and main channels. 
     
     
         42 . The device of  claim 30 , wherein said substrate further comprises at least one detection module downstream from and in fluid communication with said coalescence module, said detection module comprising a detection apparatus for evaluating the contents and/or characteristics of the nanoreactor. 
     
     
         43 . The device of  claim 42 , wherein said detection apparatus comprises an optical detector. 
     
     
         44 . The device of  claim 30 , wherein said substrate further comprises a sorting module downstream from and in fluid communication with said detection module, said sorting module comprising a sorting apparatus adapted to direct said nanoreactor into or away from a collection module in response to the contents or characterization of the nanoreactor. 
     
     
         45 . The device of  claim 30 , wherein said substrate further comprises a mixing module in fluid communication with the main channel, wherein the mixing module is downstream of the coalescence module and upstream of the detection module. 
     
     
         46 . The device of  claim 30 , wherein said substrate further comprises a delay module in fluid communication with the main channel, wherein the delay module is downstream of the coalescence module and upstream of the detection module. 
     
     
         47 . The device of  claim 30 , wherein said substrate further comprises a UV-release module in fluid communication with the main channel, wherein the UV-release module is downstream of the inlet module and upstream of the coalescence module. 
     
     
         48 . The device of  claim 30 , wherein said substrate further comprises a collection module connected to a means for storing a sample from said device. 
     
     
         49 . The device of  claim 30 , wherein said substrate further comprises a waste module connected to a means for collecting a sample discarded from said device. 
     
     
         50 . A method of creating a nanoreactor, the method comprising:
 a) providing a microfabricated substrate comprising a plurality of electrically addressable channel bearing microfluidic modules integrally arranged on said substrate so as to be in fluid communication with each other, thereby forming at least one fluorinated main channel adapted to carry at least one continuous phase fluid comprising a fluorinated oil;   b) flowing a first dispersed phase fluid comprising an aqueous solution through a first fluorinated inlet channel into the main channel such that one or more monodisperse droplets are formed in said continuous phase fluid flowing therein;   c) flowing a second dispersed phase fluid comprising an aqueous solution through a second inlet channel into the main channel such that one or more monodisperse droplets are formed in said continuous phase fluid flowing therein; and   d) coalescing at least one droplet formed in step (b) with at least one droplet formed in step (c) as the droplets pass through a coalescence module of the microfabricated substrate, thereby producing a nanoreactor, wherein the nanoreactor is about 25 μm in diameter.   
     
     
         51 . An emulsion library comprising individual specific primer-pairs to different exons and a plurality of monodisperse droplets, wherein each droplet is about 25 μm in diameter and wherein on average one specific primer-pair is comprised within each droplet, and wherein each droplet is an water-in-oil emulsion formed using a continuous phase fluid comprising a fluorocarbon oil and a surfactant and a discontinuous phase fluid comprising an aqueous fluid. 
     
     
         52 . A method of forming an emulsion library comprising:
 (a) providing individual specific primer-pairs to different exons;   (b) separately emulsifying the individual primer-pairs of step (a) in a monodisperse droplet, wherein each droplet contains on average one primer-pair; and   (c) pooling the droplets of step (b) to form a plurality of monodisperse droplets, wherein each droplet is about 25 μm in diameter;   wherein each droplet is an water-in-oil emulsion formed using a continuous phase fluid comprising a fluorocarbon oil and a surfactant and a discontinuous phase fluid comprising an aqueous fluid.