Microfluidic device and methods of using same

• US 20090176230A1
• Filed: 11/06/2008
• Published: 07/09/2009
• Est. Priority Date: 11/30/2001
• Status: Active Grant

• Current Assignee: Standard Biotools Incorporated
• Original Assignee: Fluidigm Corporation
• Inventors: McBride, Lincoln, Unger, Marc, Facer, Geoffrey, Nassef, Hany Ramez, Lucero, Michael
• Granted Patent: US 7,837,946 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• 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 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/123 Flexible; Elastomeric
• CPC Class Codes: B01L 2400/06 Valves, specific forms thereof
• CPC Class Codes: B01L 3/00 Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
• 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 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/50851 specially adapted for heating or cooling samples laboratory heating apparatus B01L7/00; incubators C12M
• 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 60/08 Integrated apparatus specially adapted for both creating and screening libraries
• CPC Class Codes: C40B 60/14 : for creating libraries
• 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 436/25 : including sample preparation

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 fluidic channel having a first end and a second end in fluid communication with each other through said channel;

an inlet in communication with said channel, said inlet for introducing a sample fluid; and

,

a valve, said valve, when actuated, causing said first channel end to become fluidically separated from said second channel end;

wherein when said sample fluid is introduced into said fluidic channel, and said valve is actuated, said sample fluid is divided into separate sample fluid portions.

2. The microfluidic device of claim 1, further comprising an outlet in communication with said channel, said outlet for removing some or all of said sample fluid.

3. The microfluidic device of claim 2, further comprising a second valve, said second valve, when actuated, causing said outlet and said fluidic channel to be in fluidic isolation.

4. The microfluidic device of claim 1, wherein said sample fluid is introduced into said fluidic channel by blind filling.

5. The microfluidic device of claim 3, wherein said sample fluid is introduced into said fluidic channel by blind filling while said second valve is actuated.

6. The microfluidic device of claim 3, wherein said second valve, when de-actuated, permits removal of some or all of said sample fluid from said fluidic channel.

7. A method of analyzing a sample fluid for a reaction to a condition comprising the steps of:

(i) providing a microfluidic device, said microfluidic device comprising;

a fluidic channel having a first end and a second end in fluid communication with each other through said channel;

an inlet in communication with said channel, said inlet for introducing said sample fluid; and

,

a valve, said valve, when actuated, causing said first channel end to become fluidically separated from said second channel end;

wherein when said sample fluid is introduced into said fluidic channel, and said valve is actuated, said sample fluid is divided into separate sample fluid portions;

(ii) introducing said sample fluid into said inlet, said sample fluid filling said first and second channel ends;

(iii) actuating said valve to separate said sample fluid into said separate portions within said first and second channel ends;

(iv) exposing said sample fluid to said conditions;

(v) analyzing said sample fluid to determine whether said conditions produced said reaction.

8. The method of claim 7, wherein said microfluidic device further comprises an outlet in communication with said channel, said outlet for removing some or all of said sample fluid, and said method further comprises a step for removing said sample fluid from said fluidic channel after said exposing step.

9. The method of claim 8, wherein said microfluidic device further comprises a second valve, said second valve, when actuated, causing said outlet and said fluidic channel to be in fluidic isolation, and said method further comprises actuating said valve prior to said introducing step to blind fill said sample fluid into said fluidic channel.

10. The method of claim 7, wherein said sample fluid is introduced into said fluidic channel by blind filling.

11. The method of claim 7, 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.

12. The method of claim 7, wherein said PCR is digital PCR.

13. The method of claim 11, wherein said reaction produces two different color results.