Microfluidic Device and Methods of Using Same

• US 20110053784A1
• Filed: 08/17/2010
• Published: 03/03/2011
• Est. Priority Date: 11/30/2001
• Status: Active Grant

• Current Assignee: Standard Biotools Incorporated
• Original Assignee: Fluidigm Corporation
• Inventors: Yi, Yong, Unger, Marc, Manger, Ian D., Miyashita-Lin, Emily, Wienecke, Anja, Facer, Geoffrey, Lucero, Michael
• Granted Patent: US 8,163,492 B2
• Time in Patent Office: Days
• US Class Current: 506/7
• CPC Class Codes: B01F 33/30 Micromixers
• CPC Class Codes: B01J 2219/00337 Valves
• CPC Class Codes: B01J 2219/00353 Pumps
• CPC Class Codes: B01J 2219/00403 in multiple arrangements
• CPC Class Codes: B01J 2219/00495 Means for heating or cooling the reaction vessels
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/0072 Organic compounds
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/06 Valves, specific forms thereof
• CPC Class Codes: B01L 3/502707 characterised by the manufacture of the container or its components by shaping or joining plastic parts B29C59/00, B29C65/00; by laminating B32B37/00; manufacture of microstructural devices in general B81C
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• CPC Class Codes: C12M 1/38 Temperature-responsive control
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 1/6844 Nucleic acid amplification reactions
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: F16K 11/20 operated by separate actuating members with one plug turning in another F16K11/12
• CPC Class Codes: G01N 2035/00366 : Several different temperatures used
• CPC Class Codes: G01N 35/00029 : provided with flat sample substrates, e.g. slides G01N35/028 takes precedence
• CPC Class Codes: G01N 35/0098 : involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C
• CPC Class Codes: Y10T 137/2076 : Utilizing diverse fluids
• CPC Class Codes: Y10T 137/2496 : Self-proportioning or correlating systems
• CPC Class Codes: Y10T 137/86292 : System with plural openings, one a gas vent or access opening
• CPC Class Codes: Y10T 137/87249 : Multiple inlet with multiple outlet
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

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.

Claims

1. A microfluidic device, comprising:

(a) a flow channel formed within an elastomeric material;

(b) a plurality of blind flow channels in fluid communication with the flow channel, with a region of each blind flow channel defining a reaction site.

4. The microfluidic device of claim 1, further comprising one or more reagents deposited within each of the reaction sites.

2-3. -3. (canceled)

5-13. -13. (canceled)

15-24. -24. (canceled)

25. A microfluidic device comprising a reaction site formed within an elastomeric substrate at which a reagent for conducting a reaction is immobilized.

26. The microfluidic device of claim 25, wherein the reagent comprises one or more reagents for conducting a nucleic acid amplification reaction.

27. The microfluidic device of claim 26, wherein the reagent comprises a primer, polymerase and/or one or more nucleotides.

28-33. -33. (canceled)

34. A method for conducting an analysis, comprising:

(a) providing a microfluidic device, comprising

(i) a flow channel formed within an elastomeric material; and

(ii) a plurality of blind flow channels in fluid communication with the flow channel, with an end region of each blind flow channel defining a reaction site;

(b) introducing at least one reagent into each of the reaction sites; and

(c) detecting a reaction at one or more of the reaction sites.

35. The method of claim 34, further comprising heating the at least one reagent within the reaction sites.

36. The method of claim 34, wherein the at least one reagent comprises a reaction mixture.

37. The method of claim 36, wherein the reaction mixture comprises the components for a nucleic acid amplification reaction and the method further comprises thermocycling the reaction mixture.

38. The method of claim 36, wherein the reaction mixture comprises the components for an enzyme assay.

39-51. -51. (canceled)

52. A method for conducting an analysis, comprising:

(a) providing a microfluidic device comprising one or more reaction sites, each reaction site comprising a first reagent for conducting an analysis that is non-covalently deposited on an elastomeric substrate;

(b) introducing a second reagent into the one or more reaction sites, whereby the first and second reagents mix to form a reaction mixture; and

(c) detecting a reaction between the first and second reagents at one or more of the reaction sites.

53. The method of claim 52, further comprising heating the reaction mixture.

54. The method of claim 52, wherein introduction of the second reagent causes the first reagent to be suspended at the reaction site.

55. The method of claim 52, wherein the one or more reaction sites is an array of reaction sites.

56. The method of claim 52, wherein the analysis is a nucleic acid analysis, the first reagent is a reagent that interacts with a nucleic acid and the second reagent is a test nucleic acid.

57. The method of claim 56, wherein the analysis is a nucleic acid amplification reaction, the first reagent comprises one or more reagents for conducting the nucleic acid amplification reaction and the second reagent is a nucleic acid template, and the method further comprises thermocycling the first and second reagents.

58-91. -91. (canceled)