Moving microdroplets in a microfluidic device

• US 7,323,140 B2
• Filed: 02/15/2002
• Issued: 01/29/2008
• Est. Priority Date: 03/28/2001
• Status: Expired due to Term

• Current Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Original Assignee: HandyLab Incorporated (Becton, Dickinson & Co)
• Inventors: Handique, Kalyan, Parunak, Gene
• Primary Examiner(s): SINES, BRIAN J
• Application Number:
  US10/075,371
• Publication Number:
  US 20020142471A1
• Time in Patent Office: 2,174 Days
• Field of Search: 422/68.1, 422/81, 422/82, 422/100, 422/101, 422/102, 422/103, 422/104, 422/939, 422/942, 422/946, 422/948, 422/50, 422/55, 422/58, 436/43, 436/52, 436/53, 436/63, 436/180, 204/193, 204/403.01, 204/403.02, 204/403.03, 435/283.1, 435/286.6, 435/286.5, 435/287.1, 435/288.4, 435/288.5, 435/288.1, 73/10.1, 73/10.2, 73/531.1, 73/15.7
• US Class Current: 422/68.1
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/0684 Venting, avoiding backpressure, avoid gas bubbles
• 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 2200/143 Quality control, feedback systems
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/1805 Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
• CPC Class Codes: B01L 2300/1827 using resistive heater
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0633 with moving parts
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 2400/0694 vents used to stop and induce flow, backpressure valves
• CPC Class Codes: B01L 2400/084 Passive control of flow resistance
• 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 3/502784 : 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 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: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: F04B 19/006 : Micropumps F04B43/043 and F04B43/095 take precedence
• CPC Class Codes: F04B 19/24 : Pumping by heat expansion of pumped fluid
• CPC Class Codes: G01N 1/4077 : by other techniques involving separation of suspended solids
• CPC Class Codes: G01N 2035/1044 : Using pneumatic means
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/118339 : with formation of a segmented stream
• CPC Class Codes: Y10T 436/25 : including sample preparation
• CPC Class Codes: Y10T 436/25375 : Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/2575 : Volumetric liquid transfer

Abstract

The present invention relates to a system and method for moving samples, such as fluid, within a microfluidic system using a plurality of gas actuators for applying pressure at different locations within the microfluidic. The system includes a substrate which forms a fluid network through which fluid flows, and a plurality of gas actuators integral with the substrate. One such gas actuator is coupled to the network at a first location for providing gas pressure to move a microfluidic sample within the network. Another gas actuator is coupled to the network at a second location for providing gas pressure to further move at least a portion of the microfluidic sample within the network. A valve is coupled to the microfluidic network so that, when the valve is closed, it substantially isolates the second gas actuator from the first gas actuator.

Claims

1. A microfluidic device, comprising:

a microfluidic network, wherein the microfluidic network comprises several modules, including;

an enrichment module configured to accept a particle-containing fluid sample, and to create an enriched sample therefrom;

a microdroplet preparation module configured to receive the enriched sample and to create a microdroplet therefrom;

a cell lysing module configured to lyse the microdroplet of enriched sample;

a mixing module configured to mix one or more reagents and the enriched sample; and

a DNA manipulation module configured to receive the enriched sample mixed with one or more reagents;

wherein the several modules are operatively connected to one another by one or more channels; and

wherein at least one of the several modules is isolatable by a valve; and

wherein at least one actuator is configured to move a microdroplet of fluid from one of the several modules to another of the several modules.

2. The microfluidic device of claim 1, further comprising:

a lower substrate having a base and an oxide layer, wherein the oxide layer contains a plurality of resistive heaters; and

an upper substrate having a bottom surface, wherein the bottom surface is bonded to the oxide layer on the lower substrate, and wherein the bottom surface includes the microfluidic network.

3. The microfluidic device of claim 2, wherein the base is made of glass.

4. The microfluidic device of claim 2, wherein the upper substrate is composed of a material selected from the group consisting of:

glass, silicon, ceramic, plastic, quartz.

5. The microfluidic device of claim 2, wherein the silicon oxide layer prevents each of the plurality of resistive heaters from directly contacting the fluid in the microfluidic network.

6. The microfluidic device of claim 1, wherein at least one of the one or more channels has a micro-scale dimension.

7. The microfluidic device of claim 6 wherein the microscale dimension is less than 75 μ

m.

8. The microfluidic device of claim 6 wherein the microscale dimension is less than 250 μ

m.

9. The microfluidic device of claim 2, wherein each heater of the plurality of resistive heaters is connected via electric leads to a pair of terminals exposed at the edge of the lower substrate.

10. The microfluidic device of claim 9, further configured to accept electrical control signals from a data acquisition and control board, wherein the control board is electrically connected to the terminals.

11. The microfluidic device of claim 10 wherein the data acquisition and control board is configured to receive instructions from a computer to apply a voltage across the pair of terminals and to cause a current to flow through the electric leads, thereby activating one or more of the resistive heaters.

12. The microfluidic device of claim 1, further comprising a sample input module, and a reagent input module, respectively configured to permit sample and reagent materials to be introduced into the device.

13. The microfluidic device of claim 12, wherein at least one of the sample input module and the reagent input module is configured to accept material input from a computer controlled laboratory robot.

14. The microfluidic device of claim 1, wherein the sample is a fluid containing whole cells.

15. The microfluidic device of claim 14, wherein the cells in the sample are bacteria, animal or human cells.

16. The microfluidic device of claim 1, wherein the sample comprises intracellular material.

17. The microfluidic device of claim 1, further comprising an input module, wherein the enrichment module is configured to accept the particle-containing fluid sample from the input module.

18. The microfluidic device of claim 17, wherein the enrichment module is configured to concentrate particles in the particle-containing fluid sample, thereby creating an enriched sample having a substantially higher ratio of particles per volume of fluid than a corresponding ratio of the particle-containing fluid sample.

19. The microfluidic device of claim 1, wherein the enrichment module comprises:

a flow-through member;

an enrichment chamber adjacent the flow-through member;

a downstream channel connecting the flow-through member to the microdroplet preparation module;

an actuator configured to force enriched sample from the enrichment chamber to the downstream channel;

a valve disposed between the actuator and the flow-through member; and

a valve disposed between the flow-through member and the downstream channel,

wherein the enrichment module is configured to allow fluid of a particle-containing fluid to pass through the flow-through member thereby accumulating the enriched sample, comprising particles of the particle-containing fluid, in the enrichment chamber.

20. The microfluidic device of claim 19, wherein the flow-through member comprises:

a first surface adjacent the enrichment chamber; and

a second surface spaced apart from the enrichment chamber and adjacent a self-contained space, wherein the space contains an absorbent material.

21. The microfluidic device of claim 19, wherein the flow-through member comprises a material having pathways smaller than the diameter of particles in the sample to be enriched.

22. The microfluidic device of claim 19, wherein the actuator is a thermally actuated gas actuator.

23. The microfluidic device of claim 22, wherein the thermally actuated gas actuator comprises a resistive heater located beneath a chamber in the microfluidic network.

24. The microfluidic device of claim 23, wherein the resistive heater is in thermal contact with the chamber, and wherein the chamber contains a volume of gas.

25. The microfluidic device of claim 22, wherein the actuator is integral with the upper substrate.

26. The microfluidic device of claim 1, wherein the microdroplet preparation module comprises:

a channel;

a positioning element situated in the channel;

a valve in communication with the channel; and

a gas actuator in communication with the channel, and configured to split a microdroplet from a fluid sample situated in the channel.

27. The microfluidic device of claim 26, wherein the positioning element is a non-wetting positioning element, or an active fluid positioning element.

28. The microfluidic device of claim 1, wherein the microdroplet preparation module is configured to accept enriched sample from the enrichment module, and is further configured to send a microdroplet of enriched sample to the cell lysing module.

29. The microfluidic device of claim 1, wherein the cell lysing module comprises:

a lysing zone;

a lysing mechanism within the lysing zone; and

a positioning element positioned upstream from the lysing zone.

30. The microfluidic device of claim 29, wherein the positioning element is selected from the group consisting of:

a non-wetting positioning element;

a capillary-assisted positioning element;

a vented positioning element; and

an active fluid positioning element.

31. The microfluidic device of claim 29, wherein the lysing mechanism comprises:

one or more electrodes that communicate with a digital acquisition and control board, and configured to deliver a pulsed electric field to a microdroplet containing cells, wherein the microdroplet is in contact with the electrodes.

32. The microfluidic device of claim 1, wherein the cell lysing module is configured to accept a microdroplet of enriched sample from the enrichment module, and is further configured to deliver a lysed microdroplet to the mixing module.

33. The microfluidic device of claim 1, wherein the cell lysing module comprises:

a lysing zone;

a lysing mechanism within the lysing zone;

an actuator disposed upstream of the lysing zone; and

a positioning element positioned downstream from the lysing zone.

34. The microfluidic device of claim 1, wherein the mixing module comprises:

a first channel; and

a second channel, adjoined to the first channel.

35. The microfluidic device of claim 34, wherein the first channel is configured to accept a lysed microdroplet from the microdroplet preparation module.

36. The microfluidic device of claim 35, wherein the sample comprises intracellular material or whole cells, and wherein the lysed microdroplet comprises polynucleotides.

37. The microfluidic device of claim 34, further comprising:

a reagent input module;

wherein the second channel of the mixing module is configured to accept reagent from the reagent input module.

38. The microfluidic device of claim 37, wherein the reagent input module comprises:

a reagent source channel, in communication with the second channel;

an input port configured to accept reagent, and to deliver a microdroplet of the reagent to the reagent source channel;

a valve in communication with the reagent source channel;

a gas actuator in communication with the reagent source channel via an opening, and

configured to split the microdroplet from the reagent when situated in the reagent source channel; and

a positioning element situated in the reagent source channel downstream from the actuator.

39. The microfluidic device of claim 38, wherein the reagent input module further comprises an overflow channel adjoining the reagent source channel.

40. The microfluidic device of claim 38, wherein the gas actuator in communication with the reagent source channel includes a second positioning element configured to prevent reagent from entering.

41. The microfluidic device of claim 38, wherein the opening and the positioning element are situated at a distance from one another that ensures that the microdroplet of reagent has a predetermined volume suitable for mixing with a lysed microdroplet of sample.

42. The microfluidic device of claim 38, wherein the positioning element is selected from a group consisting of:

a non-wetting positioning element;

a capillary-assisted positioning element;

a vented positioning element; and

an active fluid positioning element.

43. The microfluidic device of claim 1, wherein the DNA manipulation module comprises:

a DNA manipulation zone configured to accept a microdroplet of mixed lysed sample and reagent from the mixing module;

a vent configured to prevent pressure from increasing in the DNA manipulation zone during introduction of the mixed lysed sample reagent therein;

a first valve; and

a second valve;

wherein the first and second valves are positioned to prevent materials from exiting the DNA manipulation zone, when closed, and wherein the DNA manipulation zone is configured with heat sources.

44. The microfluidic device of claim 43, further comprising a detector in communication with DNA manipulation zone.

45. The microfluidic device of claim 43, wherein the DNA manipulation module is configured to carry out one or more of restriction, digestion, ligation, hybridization, or amplification of DNA material in the microdroplet of mixed lysed sample and reagent.

46. The microfluidic device of claim 43, wherein the heat sources are under control of computer to allow thermal cycling of the DNA manipulation zone.

47. The microfluidic device of claim 43, further comprising a light source that introduces light into the DNA manipulation zone.

48. The microfluidic device of claim 2 wherein both the lower substrate and the upper substrate are composed of plastic.

49. The microfluidic device of claim 48, wherein the upper substrate and microfluidic network are formed by injection moulding.

50. The microfluidic device of claim 18, wherein the ratio of particles per volume of fluid in the enriched sample is about 25 times higher than the ratio of particles per volume of fluid in the particle-containing fluid sample.

51. The microfluidic device of claim 18, wherein the ratio of particles per volume of fluid in the enriched sample is about 250 times higher than the ratio of particles per volume of fluid in the particle-containing fluid sample.

52. The microfluidic device of claim 18, wherein the ratio of particles per volume of fluid in the enriched sample is about 1,000 times higher than the ratio of particles per volume of fluid in the particle-containing fluid sample.

53. A microfluidic cartridge containing the microfluidic device of claim 1.