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Kafe 바로가기국가/구분 | United States(US) Patent 등록 |
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국제특허분류(IPC7판) |
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출원번호 | UP-0133805 (2005-05-19) |
등록번호 | US-7833708 (2011-01-16) |
발명자 / 주소 |
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출원인 / 주소 |
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대리인 / 주소 |
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인용정보 | 피인용 횟수 : 120 인용 특허 : 250 |
The present invention provides microfluidic devices and methods using the same in various types of thermal cycling reactions. Certaom devices include a rotary microfluidic channel and a plurality of temperature regions at different locations along the rotary microfluidic channel at which temperature
The present invention provides microfluidic devices and methods using the same in various types of thermal cycling reactions. Certaom devices include a rotary microfluidic channel and a plurality of temperature regions at different locations along the rotary microfluidic channel at which temperature is regulated. Solution can be repeatedly passed through the temperature regions such that the solution is exposed to different temperatures. Other microfluidic devices include an array of reaction chambers formed by intersecting vertical and horizontal flow channels, with the ability to regulate temperature at the reaction chambers. The microfluidic devices can be used to conduct a number of different analyses, including various primer extension reactions and nucleic acid amplification reactions.
What is claimed is: 1. A microfluidic device, comprising (a) a substrate; (b) a first plurality of flow channels formed within the substrate, each of the first plurality of flow channels parallel to each other; (c) a second plurality of flow channels formed within the substrate, each of the second
What is claimed is: 1. A microfluidic device, comprising (a) a substrate; (b) a first plurality of flow channels formed within the substrate, each of the first plurality of flow channels parallel to each other; (c) a second plurality of flow channels formed within the substrate, each of the second plurality of flow channels parallel to each other and in fluid communication with an inlet, the second plurality of flow channels orthogonally intersecting the first plurality of flow channels to define an array of reaction chambers; (d) isolation valves selectively actuatable to regulate solution flow to the reaction chambers, and to block flow between reaction chambers along at least one of: a flow channel in the first plurality of flow channels; or a flow channel in the second plurality of flow channels; (e) a plurality of temperature regions located along each of the second plurality of flow channels; and (f) a temperature controller operatively disposed to regulate temperature at one or more of the temperature regions. 2. The microfluidic device of claim 1, wherein the microfluidic device further comprises at least one isolation valve disposed within each of the first and second plurality of flow channels to regulate solution flow therethrough. 3. The microfluidic device of claim 2, wherein a set of the first plurality of flow channels are in fluid communication with a shared inlet such that sample introduced into the shared inlet flows into each of the flow channels of the set. 4. The microfluidic device of claim 3, wherein the set of flow channels that are in fluid communication with the shared inlet includes every other flow channel of the first plurality of flow channels. 5. The microfluidic device of claim 1, further comprising one or more control channels, and wherein one or more of the isolation valves comprises a membrane that separates the flow channel upon which it acts from one of the control channels, the membrane able to be deflected into or retracted from the flow channel upon which it acts in response to an actuation force. 6. The microfluidic device of claim 5, wherein the isolation valves comprise at least one first isolation valve and at least one second isolation valve, the first and second isolation valve having differing activation thresholds. 7. The microfluidic device of claim 6, wherein the membrane of the first isolation valve is wider than the membrane of the second isolation valve. 8. The microfluidic device of claim 6, wherein the membrane of each of the first and second valves is located at an intersection formed between one of the control channels and the flow channel upon which the first or second valve acts; and the control channel at the intersection at which the first valve is located is wider than the control channel at the intersection at which the second valve is located. 9. The microfluidic device of claim 6, wherein each of the reaction chambers disposed along at least one of the plurality of second flow channels are separated from one another by at least two first isolation valves or at least two second isolation valves. 10. The microfluidic device of claim 1, wherein the plurality of temperature regions each comprise one of the reaction chambers. 11. The microfluidic device of claim 1, wherein a single temperature controller regulates temperature at each of the reaction chambers. 12. The microfluidic device of claim 1, wherein the temperature controller is one of a plurality of temperature controllers, each temperature controller separately regulating temperature at a different reaction chamber. 13. The microfluidic device of claim 1, wherein the temperature controller is selected from the group consisting of a Peltier device, a resistive heater, a heat exchanger and an indium tin oxide element. 14. The microfluidic device of claim 1, wherein the temperature controller comprises a microfluidic chamber adapted to contain heated fluid and configured to be placed adjacent a face of the substrate. 15. The microfluidic device of claim 14, wherein the temperature controller comprises (i) a pair of spaced apart plates that are joined to one another by a fluid tight separation material located along the periphery of the plates, the space between the plates defining the microfluidic chamber, and the separation material defining the sides of the temperature controller; (ii) at least one inlet through which fluid can be introduced into the microfluidic chamber; and (iii) at least one outlet through which fluid can exit the microfluidic chamber, the outlet being located at a second end region opposing a first end region. 16. The microfluidic device of claim 1, further comprising at least two pumps, one pump operatively connected to the first plurality of flow channels to transport solution therethrough and a second pump operatively connected to the second plurality of flow channels to transport solution therethrough. 17. The microfluidic device of claim 1, wherein a polymerase enzyme is immobilized within one or more of the reaction chambers. 18. The microfluidic device of claim 1, wherein one or more nucleic acids are immobilized within one or more of the reaction chambers. 19. A method for conducting an analysis, the method comprising: (a) providing a microfluidic device, comprising (i) a substrate; (ii) a first plurality of flow channels formed within the substrate, each of the first plurality of flow channels parallel to each other; (iii) a second plurality of flow channels formed within the substrate, each of the second plurality of flow channels parallel to each other and in fluid communication with an inlet, the second plurality of flow channels orthogonally intersecting the first plurality of flow channels to define an array of reaction chambers; and (iv) isolation valves selectively actuatable to regulate solution flow to the reaction chambers, and to block flow between reaction chambers along at least one of: a flow channel in the first plurality of flow channels; or a flow channel in the second plurality of flow channels; (b) introducing a sample and one or more reactants into the reaction chambers by selective actuation of one or more of the isolation valves, whereby reaction between the sample and the one or more reactants can occur; and (c) heating regions of the microfluidic device to promote reaction between the sample and the one or more reactants within the reaction chambers. 20. The method of claim 19, wherein introducing comprises pumping sample and/or the one or more reactants into the reaction chambers for reaction. 21. The method of claim 19, wherein introducing comprises introducing the sample and the one or more reactants into the first and second plurality of flow channels and then actuating the isolation valves to allow the sample and the one or more reactants to mix by diffusion within the reaction chambers. 22. The method of claim 19, wherein the microfluidic device further comprises at least one isolation valve disposed within each of the first and second plurality of flow channels to regulate solution flow therethrough. 23. The method of claim 22, wherein a set of the first plurality of flow channels are in fluid communication with a shared inlet, the flow channels within the set including every other flow channel of the first plurality of flow channels; introducing comprises introducing the sample into the flow channels that are not within the set of flow channels and introducing the one or more reactants into the shared inlet, whereby flow channels containing samples alternate with flow channels containing reactant. 24. The method of claim 23, wherein introducing further comprises selectively actuating the isolation valves to allow sample and reactants in adjacent flow channels to mix by diffusion. 25. The method of claim 24, wherein the sample is a sample containing a nucleic acid and the one or more reactants is polymerase. 26. The method of claim 19, wherein the microfluidic device is positioned in a holder that is configured to form an air tight chamber over an inlet to each of the first plurality of flow channels; and introducing comprises placing the sample and/or the one or more reactants into the inlets to the first plurality of flow channels and then pressurizing the airtight chamber. 27. The method of claim 19, wherein heating comprises exposing the reaction chambers to a temperature cycle to promote reaction between the sample and the one or more reactants within the reaction chambers. 28. The method of claim 19, wherein heating is controlled by a temperature controller, a single temperature controller regulating the temperature at each of the reaction chambers. 29. The method of claim 19, wherein heating is controlled by a plurality of temperature controllers, different controllers regulating the temperature at different reaction chambers. 30. The method of claim 19, wherein introducing comprises introducing a sample into each of the first plurality of flow channels and introducing a reactant into each of the second plurality of flow channels. 31. The method of claim 30, wherein the sample comprises a target nucleic acid and the one or more reactants comprise one or more components for conducting a nucleic acid amplification reaction. 32. The method of claim 22, wherein the sample introduced comprises a nucleic acid and the reactant introduced comprises a component of a nucleic acid amplification or sequencing reaction. 33. The method of claim 31, wherein heating comprises exposing the target nucleic acid and the one or more components to a temperature cycle such that an amplified product is formed within the reaction chambers. 34. The method of claim 33, further comprising detecting amplified product formed with the reaction chambers. 35. The method of claim 34, wherein the amplified product bears a detectable label and the detecting step comprises detecting the label. 36. The method of claim 35, wherein the label is selected from the group consisting of a fluorophore, a chromophore, a radioisotope, a luminescent agent, a mass label, an enzyme conjugated to a nucleic acid, and a magnetic agent. 37. The method of claim 34, wherein the detecting step comprises contacting the amplified product with a label such that the amplified product becomes labeled. 38. The method of claim 37, wherein the label is a interchelating dye. 39. The method of claim 37, wherein the label is a molecular beacon. 40. The method of claim 34, wherein the amplified product is detected by conducting a quantitative PCR assay. 41. The method of claim 34, wherein detecting comprises measuring capacitance of a solution containing amplified product. 42. The method of claim 34, wherein the sample introduced into each reaction chamber comprises a plurality of target nucleic acids and a plurality of amplified products are formed. 43. The method of claim 31, wherein a polymerase enzyme is immobilized within the reaction chambers, whereby during the introduction step, the target nucleic acid and the one or more components are brought into contact with the immobilized polymerase. 44. The method of claim 31, wherein one or more nucleic acids are immobilized within the reaction chambers, whereby during the introduction step, the sample and the one or more reactants are brought into contact with the immobilized nucleic acids. 45. The method of claim 19, wherein the sample comprises a target nucleic acid and the one or more reactants are one or more components for conducting a sequencing reaction. 46. The method of claim 19, wherein the sample comprises a target nucleic acid and the one or more reactants are one or more components for conducting a quantitative PCR analysis. 47. A microfluidic device, comprising (a) a substrate (b) a first plurality of flow channels formed within the substrate, each of the first plurality of flow channels parallel to each other; (c) a second plurality of flow channels formed within the substrate, each of the second plurality of flow channels parallel to each other and in fluid communication with an inlet, the second plurality of flow channels orthogonally intersecting the first plurality of flow channels to define an array of reaction chambers; and (c) isolation valves selectively actuatable to regulate solution flow to the reaction chambers, and to block flow between reaction chambers alone at least one of: a flow channel in the first plurality of flow channels; or a flow channel in the second plurality of flow channels; wherein the isolation valves comprise a first and second isolation valve that have differing activation thresholds.
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