Microfluidic Devices and Methods of Using Same

Abstract

A variety of elastomeric-based microfluidic devices and methods for using and manufacturing such devices are provided. Certain of the devices have arrays of reaction sites to facilitate high throughput analyses. Some devices also include reaction sites located at the end of blind channels at which reagents have been previously deposited during manufacture. The reagents become suspended once sample is introduced into the reaction site. The devices can be utilized with a variety of heating devices and thus can be used in a variety of analyses requiring temperature control, including thermocycling applications such as nucleic acid amplification reactions, genotyping and gene expression analyses.

1 .- 13 . (canceled) 
     
     
         14 . A method of analyzing a sample fluid for a reaction comprising the steps of:
 (i) providing a microfluidic device, said microfluidic device comprising:
 a plurality of fluidic channels, each said channel having more than two valves associated therewith, which valves, when actuated, partition said channel into more than two reaction sites, wherein each said reaction site is delimited by exactly two actuated valves; 
 an inlet in communication with said channel, said inlet for introducing said sample fluid; and, 
 wherein when said sample fluid is introduced into said fluidic channel, and said valves are actuated, said sample fluid is divided into separate sample fluid portions; 
   (ii) introducing said sample fluid into said inlet, said sample fluid filling said channel;   (iii) actuating said valves to separate said sample fluid into more than two separate portions within said channel;   (iv) simultaneously thermocycling said separate portions of sample fluid;   (v) analyzing said separate portions of sample fluid to determine whether said thermocycling produced said reaction.   
     
     
         15 . The method of  claim 14 , wherein said sample fluid is introduced into said fluidic channel by blind filling. 
     
     
         16 . The method of  claim 14 , wherein said sample further comprises an amplifiable nucleic acid target and said conditions are polymerase chain reaction (PCR) conditions, and said reaction results in a PCR product being formed. 
     
     
         17 . The method of  claim 16 , wherein said PCR is digital PCR. 
     
     
         18 . The method of  claim 16 , wherein said reaction produces two different color results. 
     
     
         19 . The method of  claim 14  wherein the method comprises providing a microfluidic device wherein said blind channel is a branched partitioning channel system and said microfluidic device further comprises a plurality of valves and wherein said sample is introduced into said branched partitioning system and said plurality of valves are actuated to isolate said sample into a plurality of isolated reaction volumes. 
     
     
         20 . The method of  claim 14  wherein the analysis is single nucleotide polymorphism analysis. 
     
     
         21 . The method of  claim 14  wherein the analysis is genotyping. 
     
     
         22 . The method of  claim 14  wherein the analysis is real time PCR. 
     
     
         23 . The method of  claim 14  wherein each reaction volume is less than 20 μL. 
     
     
         24 . The method of  claim 14  wherein each reaction volume is less than 10 μL. 
     
     
         25 . The method of  claim 14  wherein said sample is partitioned into at least 500 isolated reaction volumes.