Integrated active flux microfluidic devices and methods

Abstract

The invention relates to a microfabricated device for the rapid detection of DNA, proteins or other molecules associated with a particular disease. The devices and methods of the invention can be used for the simultaneous diagnosis of multiple diseases by detecting molecules (e.g. amounts of molecules), such as polynucleotides (e.g., DNA) or proteins (e.g., antibodies), by measuring the signal of a detectable reporter associated with hybridized polynucleotides or antigen/antibody complex. In the microfabricated device according to the invention, detection of the presence of molecules (i.e., polynucleotides, proteins, or antigen/antibody complexes) are correlated to a hybridization signal from an optically-detectable (e.g. fluorescent) reporter associated with the bound molecules. These hybridization signals can be detected by any suitable means, for example optical, and can be stored for example in a computer as a representation of the presence of a particular gene. Hybridization probes can be immobilized on a substrate that forms part of or is exposed to a channel or channels of the device that form a closed loop, for circulation of sample to actively contact complementary probes. Universal chips according to the invention can be fabricated not only with DNA but also with other molecules such as RNA, proteins, peptide nucleic acid (PNA) and polyamide molecules.

1 . A microfluidic device comprising:
 a loop channel communicating with at least one service channel,   a microvalve separating the loop channel from the service channel, and   a pump associated with the loop channel;   wherein the pump is a peristaltic pump comprising at least three cooperating microvalves acting within the loop channel.   
     
     
         2 .- 62 . (canceled) 
     
     
         63 . A microfluidic device according to claim  62 , wherein
 the substrate is a microtiter plate having microtiter wells, each microtiter well having a target molecule patterned thereon; and   the microtiter plate is connected to the treatment layer so that at least a portion of the length of each loop channel is sealed by a microtiter well.   
     
     
         64 - 69 . (canceled) 
     
     
         70 . A method for mixing two or more different fluids in a microfluidic device, which method comprises:
 (a) introducing the different fluids to a microfluidic device, the microfluidic device having:   (i) a loop channel communicating with at least one service channel,   (ii) a microvalve separating the loop channel from the service channel, and   (iii) a pump associated with the loop channel,   so that each of the different fluids is loaded into the loop channel; and   (b) activating the pump associated with the loop channel to mix the different fluids.   
     
     
         71 .- 92 . (canceled)