Microfluidic cartridge and method of making same

• US 9,815,057 B2
• Filed: 04/28/2014
• Issued: 11/14/2017
• Est. Priority Date: 11/14/2006
• Status: Active Grant

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Handique, Kalyan
• Primary Examiner(s): FORMAN, BETTY J
• Application Number:
  US14/263,208
• Publication Number:
  US 20140323357A1
• Time in Patent Office: 1,296 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/142 Preventing evaporation
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2300/1844 using fans
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0677 phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
• CPC Class Codes: B01L 2400/0694 vents used to stop and induce flow, backpressure valves
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• 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 3/502723 characterised by venting arrangements
• CPC Class Codes: B01L 3/502738 characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: B81C 1/00087 Holes
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: F16K 2099/008 : Multi-layer fabrications
• CPC Class Codes: F16K 2099/0084 : Chemistry or biology, e.g. "lab-on-a-chip" technology
• CPC Class Codes: F16K 99/0001 : Microvalves microdevices B81B1/00; manufacture or treatment of devices or systems in or on a substrate B81C1/00; microfluidic structures B01L3/5027; micropumps F04B19/006

Abstract

The present technology provides for a microfluidic substrate configured to carry out PCR on a number of polynucleotide-containing samples in parallel. The substrate can be a single-layer substrate in a microfluidic cartridge. Also provided are a method of making a microfluidic cartridge comprising such a substrate. Still further disclosed are a microfluidic valve suitable for use in isolating a PCR chamber in a microfluidic substrate, and a method of making such a valve.

Claims

1. A method of isolating a plurality of polynucleotide-containing samples on a microfluidic cartridge, the method comprising:

introducing a first polynucleotide-containing sample into a first reaction chamber via a first inlet port, the first inlet port in fluid communication with the first reaction chamber and a first outlet;

introducing a second polynucleotide-containing sample into a second reaction chamber via a second inlet port, the second inlet port in fluid communication with the second reaction chamber and a second outlet, wherein the first polynucleotide-containing sample is different than the second polynucleotide-containing sample;

thermally actuating a first set of microfluidic valves to isolate the first polynucleotide-containing sample from the first inlet port and the first outlet; and

independent of the actuation of the first set of microfluidic valves, thermally actuating a second set of microfluidic valves to isolate the second polynucleotide-containing sample from the second inlet port and the second outlet, the isolation effected by the first and the second set of microfluidic valves preventing movement of fluid into and out of the first and the second reaction chambers.

2. The method of claim 1, wherein thermally actuating the first set of microfluidic valves comprises actuating a first microfluidic valve spatially separated from the first inlet port and actuating a second microfluidic valve spatially separated from the first outlet, and wherein thermally actuating the second set of microfluidic valves comprises actuating a first microfluidic valve spatially separated from the second inlet port and actuating a second microfluidic valve spatially separated from the second outlet.

3. The method of claim 1, further comprising amplifying one or more polynucleotides in the first reaction chamber independently of amplifying one or more polynucleotides in the second reaction chamber.

4. The method of claim 1, further comprising simultaneous loading samples from a multiple-pipette head dispenser into the first inlet port and the second inlet port.

5. The method of claim 1, wherein, prior to thermally actuating the first set of microfluidic valves, the first set of microfluidic valves are initially open and in fluid communication with the first reaction chamber and wherein, prior to thermally actuating the second set of microfluidic valves, the second set of microfluidic valves are initially open and in fluid communication with the second reaction chamber.

6. The method of claim 1, further comprising amplifying the plurality of polynucleotide-containing samples after performing the method of isolating the plurality of polynucleotide-containing samples.

7. The method of claim 1, further comprising mixing each sample of the plurality of polynucleotide-containing samples with a reagent prior to performing the method of isolating the plurality of polynucleotide-containing samples.

8. The method of claim 1, wherein thermally actuating a first set of microfluidic valves further comprises heating a volume of trapped air.

9. The method of claim 1, wherein thermally actuating a first set of microfluidic valves further comprises heating a conical chamber of air.

10. A method of isolating a plurality of polynucleotide-containing samples, the method comprising:

providing a plurality of sample lanes, wherein each of the plurality of sample lanes comprises a microfluidic network having, in fluid communication with one another;

an inlet and an outlet;

a first valve and a second valve;

a reaction chamber,

a first channel leading from the inlet, via the first valve, to the reaction chamber; and

a second channel leading from the reaction chamber, via the second valve, to the outlet;

introducing a first polynucleotide-containing sample into a first reaction chamber in a first microfluidic network and introducing a second polynucleotide-containing sample into a second reaction chamber in a second microfluidic network, wherein the first polynucleotide-containing sample is different than the second polynucleotide-containing sample;

isolating the first polynucleotide-containing sample from the inlet and the outlet to prevent movement of fluid into or out of the first reaction chamber by closing the first valve and the second valve of the first microfluidic network by application of heat; and

isolating the second polynucleotide-containing sample from the inlet and the outlet to prevent movement of fluid into or out of the second reaction chamber by closing the first valve and the second valve of the second microfluidic network by application of heat, the first valve in the first microfluidic network closed independently of the first valve in the second microfluidic network.

11. The method of claim 10, wherein, in each microfluidic network, the first valve is spatially separated from the first inlet and the second valve is spatially separated from the outlet.

12. The method of claim 10, further comprising amplifying one or more polynucleotides in the first reaction chamber independently of amplifying one or more polynucleotides in the second reaction chamber.

13. The method of claim 10, wherein the first polynucleotide-containing sample and the second polynucleotide-containing sample are simultaneously introduced by a multiple-pipette head dispenser.

14. The method of claim 10, wherein the first valve and the second valve in each microfluidic network are both initially open and in fluid communication with the reaction chamber of the microfluidic network.

15. The method of claim 10, wherein isolating the first polynucleotide-containing sample and isolating the second polynucleotide-containing sample comprises closing the first and second valves in the first microfluidic network independent of closing the first and second valves in the second microfluidic network.

16. The method of claim 10, wherein a microfluidic cartridge comprises the plurality of sample lanes.

17. A method of isolating a plurality of polynucleotide-containing samples, the method comprising:

introducing a first polynucleotide-containing sample into a first inlet port, wherein the first polynucleotide-containing sample flows from the first inlet port through a first valve spaced from the first inlet port into a first amplification chamber and toward a second valve spaced from the first amplification chamber;

introducing a second polynucleotide-containing sample into a second inlet port, wherein the first polynucleotide-containing sample is different than the second polynucleotide-containing sample, wherein the second polynucleotide-containing sample flows from the second inlet port through a third valve spaced from the second inlet port into a second amplification chamber and toward a fourth valve spaced from the second amplification chamber;

applying heat to the first valve and the second valve to isolate the first polynucleotide-containing sample in the first amplification chamber;

independent of applying heat to the first valve and the second valve, applying heat to the third valve and the fourth valve to isolate the second polynucleotide-containing sample in the second amplification chamber independent of the isolation of the first polynucleotide-containing sample in the first amplification chamber.

18. The method of claim 17, wherein the first valve is spatially separated from the first inlet port, and wherein the third valve is spatially separated from the second inlet port.

19. The method of claim 17, wherein applying heat to the first and the second valves prevents movement of the first polynucleotide-containing sample into and out of the first amplification chamber.

20. The method of claim 17, further comprising amplifying one or more polynucleotides in the first reaction chamber independently of amplifying one or more polynucleotides in the second reaction chamber.

21. The method of claim 17, further comprising simultaneous loading of polynucleotide-containing samples from a multiple-pipette head dispenser into the first inlet port and the second inlet port.

22. The method of claim 17, wherein, prior to applying heat to the first and second valves, the first valve and the second valve are both initially open and in fluid communication with the first amplification chamber, and wherein, prior to applying heat to the third and fourth valves, the third valve and the fourth valve are both initially open and in fluid communication with the second amplification chamber.

23. The method of claim 17, wherein a microfluidic cartridge comprises a plurality of sample lanes, wherein a first lane comprises the first inlet port, the first valve, the first amplification chamber and the second valve, and wherein a second lane comprises the second inlet port, the third valve, the second amplification chamber and the fourth valve.