Microfluidic cartridge

• US 9,322,054 B2
• Filed: 02/21/2013
• Issued: 04/26/2016
• Est. Priority Date: 02/22/2012
• Status: Active Grant

• Current Assignee: Leidos Innovations Technology, Inc.
• Original Assignee: Lockheed Martin Corporation (Martin Marietta Corporation), Microlab Diagnostics Limited
• Inventors: Root, Brian, Scott, Orion N., South, Douglas J., Bienvenue, Joan, Kinnon, Paul, Landers, James, Saul, David, Tsuei, An-Chi, Hayes, Jason, Springer, Matthew, Solomon, Matthew, van Ruijven, Peter, Egan, Michael
• Primary Examiner(s): Wilder, Cynthia B
• Application Number:
  US13/773,350
• Publication Number:
  US 20130217026A1
• Time in Patent Office: 1,160 Days
• Field of Search: 422/417, 435/91.2
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/04 Exchange or ejection of cartridges, containers or reservoirs
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2200/148 Specific details about calibrations
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0663 Whole sensors
• CPC Class Codes: B01L 2300/0672 Integrated piercing tool
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0832 cylindrical, tube shaped
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2300/161 Control and use of surface tension forces, e.g. hydrophobic, hydrophilic
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 2300/1872 Infrared light
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/049 : vacuum
• CPC Class Codes: B01L 2400/0683 : mechanically breaking a wall or membrane within a channel or chamber
• CPC Class Codes: B01L 2400/088 : by specific surface properties
• CPC Class Codes: B01L 3/502715 : characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: B01L 3/502723 : characterised by venting arrangements
• CPC Class Codes: B01L 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502738 : characterised by integrated valves microvalves F16K99/0001
• CPC Class Codes: B01L 3/502753 : characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B01L 3/502784 : specially adapted for droplet or plug flow, e.g. digital microfluidics automatic analysis using a stream of discrete samples in a tube system G01N35/08
• CPC Class Codes: B01L 3/5029 : using swabs
• 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: C12P 19/34 : Polynucleotides, e.g. nucleic acids, oligoribonucleotides
• CPC Class Codes: C12Q 1/6844 : Nucleic acid amplification reactions
• CPC Class Codes: C12Q 1/6853 : using modified primers or templates
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: G01N 21/75 : Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated systems in which material is burnt in a flame or plasma G01N21/72, G01N21/73

Abstract

A microfluidic cartridge can include at least one nucleic acid analysis portion. Each nucleic acid analysis portion can include a fluidic network being configured for micro-liter volumes or less, a sample input at the beginning of the fluidic network, a plurality of vent ports and fluidic channels in the fluidic network configured to effectuate hydrodynamic movement within the fluidic network, an extraction mixture reservoir in the fluidic network, a mixing chamber in the fluidic network, an amplification chamber in the fluidic network, and a separation channel in the fluidic network. A nucleic acid analyzer can be capable of performing nucleic acid analysis using the microfluidic cartridge. A nucleic acid analysis method can be performed using the microfluidic cartridge.

Claims

1. A microfluidic cartridge, comprising:

at least one nucleic acid analysis portion, each nucleic acid analysis portion including;

a fluidic network defined within the nucleic acid analysis portion, the fluidic network being configured for micro-liter volumes or less, the fluidic network including at least one frangible seal formed from a depression defined in a fluidic channel with a fluid flow resistance determined by a depth and a width of the depression;

a sample input at a beginning of the fluidic network, the sample input having a fitting that is configured to be mated to a complementary fitting of a sample acceptor to form a fluid-tight seal;

a plurality of vent ports and fluidic channels in the fluidic network configured to effectuate hydrodynamic movement within the fluidic network;

an extraction mixture reservoir in the fluidic network, the extraction mixture reservoir being configured to hold an enzymatic mixture for performing nucleic acid extraction on a sample provided by the sample acceptor;

a mixing chamber in the fluidic network, the mixing chamber being configured to mix amplification reagents and a portion of an extracted nucleic acid mixture; and

an amplification chamber in the fluidic network, the amplification chamber being configured to hold an amplification mixture during nucleic acid amplification.

2. The microfluidic cartridge of claim 1, wherein at least a major portion of the microfluidic cartridge is covered by a sealing layer.

3. The microfluidic cartridge of claim 1, wherein the fluidic network comprises a separation channel configured to separate nucleic acid fragments.

4. The microfluidic cartridge of claim 1, wherein the fluidic network includes a plurality of frangible seals.

5. The microfluidic cartridge of claim 4, wherein the plurality of frangible seals include at least one frangible seal that has greater fluid flow resistance than other frangible seals.

6. The microfluidic cartridge of claim 1, wherein at least one frangible seal is formed from a portion of a sealing layer that is extended in the depression and is adhered to a base of the depression in the fluidic network.

7. The microfluidic cartridge of claim 1, wherein the sample input has a universal connector selected from the group consisting of Luer-Lok connectors, threaded connectors, and flanged connectors.

8. The microfluidic cartridge of claim 1, wherein the extraction mixture reservoir is configured to hold from 5 μ

l to 500 μ

l of fluid.

9. The microfluidic cartridge of claim 8, wherein the extraction mixture reservoir is configured to hold from 5 μ

l to 250 μ

l of fluid.

10. The microfluidic cartridge of claim 1, wherein the fluidic network includes at least one reagent reservoir configured to hold a reagent solution, the reagent reservoir being in fluid communication with bypass fluidic channels for loading the reagent solution in the reagent reservoir and being in fluid communication with fluidic channels that have frangible seals to block the reagent solution from entering or prematurely entering other portions of the fluidic network.

11. The microfluidic cartridge of claim 1, further comprising an amplification reagent reservoir in the fluidic network, the amplification reagent reservoir being configured to hold amplification reagents for performing nucleic acid amplification.

12. The microfluidic cartridge of claim 11, wherein more than one amplification reagent reservoir is in the fluidic network.

13. The microfluidic cartridge of claim 1, wherein a portion of the fluidic network is configured to move amplification reagents and the portion of the extracted nucleic acid mixture into the mixing chamber in discrete volumes through serial fluid communication.

14. The microfluidic cartridge of claim 1, wherein a portion of the fluidic network includes one-time actuation features.

15. The microfluidic cartridge of claim 14, wherein the one-time actuation features are frangible seals.

16. The microfluidic cartridge of claim 1, wherein the mixing chamber includes a liquid mixing portion and a gas vent portion.

17. The microfluidic cartridge of claim 16, wherein the liquid mixing portion is below the gas vent portion.

18. The microfluidic cartridge of claim 16, wherein the mixing chamber has a surface including a hydrophobic portion.

19. The microfluidic cartridge of claim 1, further comprising an internal control reservoir in the fluidic network, the internal control reservoir being configured to hold an internal control solution for providing an internal control during nucleic acid separation.

20. The microfluidic cartridge of claim 19, wherein the internal control solution includes an internal lane standard (ILS).

21. The microfluidic cartridge of claim 1, further comprising a sieving polymer reservoir in fluid communication with the separation channel, the sieving polymer reservoir being configured to hold a sieving polymer solution and to provide the sieving polymer solution to the separation channel.

22. The microfluidic cartridge of claim 21, wherein the sieving polymer solution comprises an alignment dye for calibrating a detection module of a nucleic acid analyzer.

23. The microfluidic cartridge of claim 21, further comprising a solution reservoir in fluid communication with the separation channel.

24. The microfluidic cartridge of claim 23, wherein the solution reservoir is configured to hold a dilution and/or buffer solution and/or urea and/or dimethyl-formamide (DMF) in water.

25. The microfluidic cartridge of claim 1, wherein the separation channel includes a detection region configured to enable laser beam detection of separated nucleic acids.

26. The microfluidic cartridge of claim 1, wherein the sample acceptor includes an input-matable portion, an acceptor portion, and a detachable portion for collecting a biological sample containing a minimum of 100 cells.