Microfluidic device and methods of using same

• US 8,163,492 B2
• Filed: 08/17/2010
• Issued: 04/24/2012
• Est. Priority Date: 11/30/2001
• Status: Expired due to Term

• Current Assignee: Standard Biotools Incorporated
• Original Assignee: Fluidigm Corporation
• Inventors: Unger, Marc, Manger, Ian D., Lucero, Michael, Yi, Yong, Miyashita-Lin, Emily, Wienecke, Anja, Facer, Geoffrey
• Primary Examiner(s): Yang, N. C.
• Application Number:
  US12/858,364
• Publication Number:
  US 20110053784A1
• Time in Patent Office: 616 Days
• Field of Search: None
• US Class Current: 435/6.12
• 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 method for conducting an analysis, comprising:

(a) introducing a solution into a first flow channel of a microfluidic device,

wherein the solution comprises a sample, a reactant, or both;

wherein the first flow channel is in fluid communication with a plurality of blind flow channels, with a region at the terminus of each blind flow channel defining a reaction site,

wherein each blind flow channel is associated with a valve that when closed isolates the reaction site from the first flow channel, and

wherein the valves associated with each of the plurality of blind flow channels in fluid communication with the first flow channel are under common control and are coordinately closed or opened,

(b) flowing said solution through said blind flow channels and into the reaction sites;

(c) closing the valves, thereby isolating the reaction sites; and

(d) detecting a reaction between the sample and the reactant at one or more of the reaction sites,

wherein said device comprises an elastomeric materials that allows air within the blind flow channels to escape through the material as the solution is flowed through the blind flow channels and into the reaction sites.

2. The method of claim 1 wherein the valves comprise elastomeric membranes that deflect into or out of the blind flow channel.

3. The method of claim 1, wherein at least one reactant is deposited in each of the reaction sites prior to step (a).

4. The method of claim 1 wherein the solution comprises at least one sample and at least one reagent.

5. The method of claim 1, further comprising heating the solution within the reaction sites after step (d).

6. The method of claim 1, wherein the solution comprises the components for a nucleic acid amplification reaction.

7. The method of claim 6 comprising thermocycling the solution within the reaction sites.

8. The method of claim 6 wherein the nucleic acid amplification reaction is a polymerase chain reaction.

9. The method of claim 1, wherein the reaction comprises the action of an enzyme on a substrate.

10. The method of claim 9 wherein the reaction is a nucleic acid amplification reaction.

11. The method of claim 1 where the reaction results in emission of a detectable signal.

12. The method of claim 1 wherein the microfluidic device comprises polydimethylsiloxane.

13. The method of claim 2, wherein there are at least 100 reaction sites.

14. The method-of claim 13, wherein there are at least 1,000 reaction sites.

15. The method of claim 14, wherein there are at least 10,000 reaction sites.

16. The method of claim 15, wherein there are at least 100,000 reaction sites.

17. The method of claim 1 wherein the fluid capacity of each isolated reaction site is less than 20 nL.

18. The method of claim 17 wherein the fluid capacity of each isolated reaction site is less than 10 nL.

19. The method of claim 2, wherein the microfluidic device comprises a deflectable membrane formed from an elastomeric material having a Young'"'"'s modulus in the range 50 Pa to 10 MPa.

20. The method of claim 2, wherein the microfluidic device comprises a deflectable membrane formed from an elastomeric material having a Young'"'"'s modulus in the range 100 Pa to 1 MPa.

21. The method of claim 1, wherein the microfluidic device comprises an array of reaction sites having a density of at least 100 sites/cm².

22. The method of claim 21, wherein the microfluidic device comprises an array of reaction sites having a density of at least 1000 sites/cm².

23. The method of claim 2 wherein the membranes are deflected in response to the same actuation force.

24. The method of claim 2, wherein the reaction between the sample and the reactant is detected by detecting a product formed by the reaction.