Sample preparation integrated chip

• US 20030138941A1
• Filed: 10/24/2002
• Published: 07/24/2003
• Est. Priority Date: 10/26/2001
• Status: Active Grant

• Current Assignee: Defence Science and Technology, Nanyang Technological University
• Original Assignee: Defence Science and Technology
• Inventors: Yap, Eric Peng Huat, Chen, Longqing, Gong, Haiqing
• Granted Patent: US 7,338,760 B2
• Time in Patent Office: Days
• US Class Current: 435/287.2
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• 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 2300/044 pierceable, e.g. films, membranes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0861 Configuration of multiple channels and/or chambers in a single devices
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2400/0403 specific forces
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0448 Marangoni flow; Thermocapillary effect
• CPC Class Codes: B01L 2400/0475 specific mechanical means and fluid pressure
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/06 Valves, specific forms thereof
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• 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: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: C12Q 1/68 involving nucleic acids
• CPC Class Codes: C12Q 2565/629 being a microfluidic device
• CPC Class Codes: G01N 1/28 Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Qmounting specimens on microscopic slides G02B21/34; means for supporting the objects or the materials to be analysed in electron microscopes H01J37/20 ; laboratory gas handling apparatus B01L5/00
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention relates to an apparatus comprising a substrate having at least one assay station. The at least one assay station has at least a first assay station channel and at least a second assay station channel and the first and second assay station channels each separately being in communication with the at least one assay station. The apparatus has an arrangement of at least first and second multipurpose channels in communication with the first and second assay station channels, respectively. The first multipurpose channel and first assay station channel have internal surface characteristics conducive to conduction of a sample solution therethrough. There is at least one sample fluid inlet in communication with the at least first multipurpose channel, and at least one isolation-medium inlet in communication with the at least first and second multipurpose channels. The at least one second multipurpose channel has an internal surface portion non-conducive to conduction of said sample solution.

Claims

1. ) An apparatus comprising:

a substrate having at least one assay station;

an arrangement of at least one first multipurpose channels and at least one second multipurpose channel wherein said at least one assay station being situated in a position intermediate between said first and second multipurpose channels and in fluid communication therewith, wherein said first multipurpose channel has at least one characteristic conducive to conduction of a sample fluid therethrough;

at least one sample fluid inlet in communication with said at least first multipurpose channel; and

at least one isolation-medium inlet in communication with said at least first and second multipurpose channels, said at least one second multipurpose channel having at least one characteristic non-conducive to conduction of said sample fluid.

2. ) The apparatus of claim 1 wherein said fluid communication is via at least first and second assay station channels in communication with said first and second multipurpose channels.

3. ) The apparatus of claim 1 wherein said first multipurpose channel characteristic conducive to conduction of said sample fluid comprises at least one of internal surface characteristic and/or shape characteristic and said at least one second multipurpose channel characteristic that is non-conducive to conduction of said sample fluid comprises at lease one of an internal surface portion and/or shape characteristics.

4. ) The apparatus according to claim 1 wherein the apparatus further comprises a sealing layer sealing at least one assay station.

5. ) The apparatus according to claim 1 wherein the internal surface of said first multipurpose channel permits flowthrough of at least one of a sample fluid, air and an isolation-medium.

6. ) The apparatus according to claim 1 wherein the internal surface of said second multipurpose channel permits the flowthrough of at least one of air or an isolation-medium.

7. ) The apparatus of claim 1 wherein at least a portion of said assay station, first multipurpose channel and second multipurpose channel are formed in the substrate layer.

8. ) The apparatus of claim 4 wherein at least a portion of at least one of said assay station, first multipurpose channel and second multipurpose channel is formed in the substrate layer and at least a portion of at least one of said assay station, first multipurpose channel and second multipurpose is formed in the sealing layer.

9. ) The apparatus according to claim 2 wherein said internal surface of said multipurpose channel and a surface of said second assay station channel immediately adjacent to the intersection of the second assay station channel and said second multipurpose channel are non-conducive to conduction of said sample fluid.

10. ) The apparatus according to claim 2 wherein said at least first and second multipurpose channels are in communication with a plurality of assay stations via the first and second assay station channels, respectively, of said plurality of assay stations.

11. ) The apparatus according to claim 10 wherein said plurality of assay stations are arranged to provide at least one of simultaneous or sequential filling of said plurality of assay stations with said sample fluid solution conducted thereto.

12. ) The apparatus according to claim 10 wherein said plurality of assay stations are arranged to provide at least one of simultaneous or sequential filling of said first and second multipurpose channels with said isolation medium to seal said plurality of assay stations.

13. ) The apparatus of claim 1 wherein said at least one assay station has disposed therein at least one reaction assay component.

14. ) The apparatus of claim 1 wherein said sample fluid inlet is in communication with a sample fluid preparation element.

15. ) The apparatus of claim 14 further comprising at least one of a sample preparation chamber and a lid.

16. ) The apparatus of claim 1 further comprising at least one element for controlling fluid flow in at least one of said channels.

17. ) The apparatus of claim 1 further comprising a chamber for introduction of flow-promoting fluid.

18. ) The apparatus of claim 17 wherein at least one of said chamber or an inlet is in communication with a mixing chamber for mixing said flow-promoting fluid with said sample fluid.

19. ) The apparatus according to claim 1 wherein said at least one assay station comprises at least one component of an assay reaction pre-loaded therein.

20. ) The apparatus according to claim 19 wherein said least one component of said assay reaction at said at least one assay station provides at least one of detectable qualitative or quantitative data.

21. ) The apparatus of claim 20 further comprising beads having said at least one component of said assay reaction.

22. ) The apparatus according to claim 19 wherein said at least one component of said assay reaction is secured to said at least one assay station.

23. ) The apparatus according to claim 15 wherein said sample preparation chamber further comprises an absorbent.

24. ) The apparatus according to claim 1 further comprising an absorbent in communication with a terminal portion of at least one of said at least first and second multipurpose channels.

25. ) The apparatus according to claim 24 wherein said absorbent is removeably attached to the terminal portion of said at least first and second multipurpose channels.

26. ) The apparatus according to claim 16 wherein said at least one flow controlling element is disposed between said sample preparation chamber and said at least first multipurpose channel.

27. ) The apparatus according to claim 16 said at least one flow controlling element is disposed adjacent said at least one assay station.

28. ) The apparatus according to claim 1 wherein said at least one assay station is further comprised of flow promoting structures.

29. ) The apparatus according to claim 2 wherein said at least a portion of said at least one first assay station channel has a cross-sectional area that is less than the cross-sectional area of at least a portion of said at least second assay station channel.

30. ) The apparatus of claim 9 wherein said first and second multipurpose channels provide a path by which said plurality of assay stations are sealed via the flow through of an isolation medium.

31. ) The apparatus according to claim 1 wherein exposed portions of the said at least first and second multipurpose channels are sealed with a solid from ambient atmosphere adhesively, mechanically, electrically, or magnetically after the first and second multipurpose channels are filled with a sample fluid and/or an isolation medium.

32. ) An apparatus for analyzing a sample fluid, comprising;

a substrate, said substrate comprising;

a sample preparation chamber for removal of undesired components from a sample fluid;

a sample fluid inlet fluidically coupled to said sample preparation chamber;

a chamber or inlet for receiving a flow-promoting fluid;

a flow controlling element for controlling flow of a sample fluid from said sample preparation chamber to said chamber or inlet for receiving a flow-promoting fluid;

at least one first multi purpose channel fluidically coupled to said chamber;

at least one assay station;

said first multi purpose channel in communication to said at least one assay station;

a second flow controlling element for isolating and permitting flow of a fluid from said chamber to said at least one assay station; and

a first assay station channel, coupled to said at least one assay station.

33. ) The apparatus of claim 32 wherein said sample preparation chamber comprises a sintered glass block which seals at least a portion of said sample preparation chamber.

34. ) The apparatus according to claim 32 wherein portions of said at least first multipurpose channel is sealed with a solid from ambient atmosphere adhesively, mechanically, electrically, or magnetically after the first multipurpose channel is filled with sample fluid and/or an isolation medium.

35. ) The apparatus of claim 33 wherein an absorbent adheres to said sintered glass block for absorbing said undesired undesired components.

36. ) The apparatus of claim 32 wherein a sealing layer seals at least one of said filtration chamber, chamber, first multipurpose channel, assay station, and first assay channel.

37. ) The apparatus of claim 32 further comprising a diaphragm which seals at least a portion of said filtration chamber from the environment;

and

a vibrating actuator for vibrating said diaphragm, thereby agitating a lysing buffer.

38. ) The apparatus of claim 32 wherein said substrate comprises at least one of (a) glass, (b) plastic, (c) elastomer, (d) a composite, (e) silicon and (f) metal.

39. ) The apparatus of claim 38 wherein said elasomer comprises poly-dimethylsiloxane.

40. ) A method for conducting reactions on a substrate wherein said a substrate comprises at least one assay station, an arrangement of at least first and second multipurpose channels wherein said at least one assay station being situated in a position intermediate between said first and second multipurpose channels and in fluid communication therewith, and wherein said first multipurpose channel has internal surface characteristics conducive to conduction of a sample solution therethrough, at least one sample fluid receiving area in communication with said at least first multipurpose channel, at least one isolation-medium inlet in communication with said at least first and second multipurpose channels, said least one second multipurpose channel having at least an internal surface portion non-conducive to conduction of said sample solution comprising:

obtaining a sample fluid;

introducing a sample fluid to at least one sample inlet;

filling said at least one assay station via said at least one multipurpose channel;

allowing isolation-medium from said at least one isolation medium port to flow into at least said first multipurpose channel; and

running at least one reaction at said at least one assay station, said reaction providing at least one of qualitative or quantitative data relating to said sample fluid.

41. ) The method according to claim 40 further comprising running said at least one reaction under temperature control.

42. ) The method according to claim 40 further comprising the step of obtaining said sample fluid from a test sample.

43. ) The method according to claim 42 further comprising the step of subjecting said test sample to at least one preparative operation.

44. ) The method according to claim 43 further comprising performing said at least one preparative operation separately from said substrate.

45. ) The method according to claim 43 further comprising performing said at least one preparative operation at at least one of upon or within said substrate.

46. ) The method according to claim 40 wherein said at least one of qualitative or quantitative data provides at least one of a colorimetric, flurometric or luminescent result.

47. ) The method according to claim 43 wherein said at least one preparative operation provides nucleic acids susceptible for use in said at least one reaction.

48. ) The method of claim 40 further comprising the step of disposing at least one assay reaction component into said at least one assay station.

49. ) The method according to claim 40 wherein said step of running said at least one reaction comprises nucleic acid amplification.

50. ) The method of claim 40 further comprising obtaining said at least one of qualitative or quantitative data utilizing fluorescence.

51. ) The method according to claim 50 wherein said fluorescence is provided by at least one of binding of a fluorophore or hybridization of fluorophore containing probe.

52. ) The method according to claim 40 wherein said qualitative or quantitative data is obtained via probe labeled with at least one of a fluorophore, an enzyme or component of a binding complex.

53. ) The method according to claim 40 further comprising the step of displacing said sample fluid via isolation-medium.

54. ) The method according to claim 52 further comprising introducing sequentially said isolation-medium into said at least first and second multipurpose channels.

55. ) The method according to claim 53 wherein said isolation medium is first introduced into said at least first multipurpose channel followed by introduction into said at least second multipurpose channel.

56. ) The method according to claims 54 or 55 wherein said introduction of isolation medium provides the purging of air from said at least second multipurpose channel and the purging of said sample fluid from said at least first multipurpose channel, resulting in the isolation of said at least one assay station containing said sample fluid.

57. ) The method according to claim 49 further comprising the step of exposing said at least one assay station to irradiation.

58. ) The method according to claim 40 further comprising the step of at least one of solidifying, curing and polymerizing said isolation medium.

59. ) The method according to claim 40 wherein exposed portions of the said at least first and second multipurpose channels are sealed with a solid from ambient atmosphere adhesively, mechanically, or magnetically after the first and second multipurpose channels are filled with sample fluid and/or isolation medium.

60. ) The method according to claim 40 further comprising heating said fluid sample prior to filling said at least one assay station.

61. ) The method according to claim 43 wherein the step of obtaining said sample fluid includes at least one preparative operation in a sample preparation chamber, comprising at least one of exposing said test sample to a lysing buffer, elution buffer and a washing buffer, in order to obtain said sample fluid.

62. ) The method according to claim 61 further comprising adding a flow promoting fluid to said sample fluid.

63. ) The method according to claim 61 wherein said at least one preparative operation is conducted upon said substrate, further comprising the step of agitating said substrate in order to promote the entry of nucleic acids, contained in a nucleic acid containing test sample, to enter into said sample fluid.

64. ) The method of claim 63 wherein said step of agitating said substrate is performed by agitating said substrate at the resonant frequency of at least one of said substrate and the sample fluid contained in said sample preparation chamber.

65. ) The method of claim 63 wherein said agitating step is performed by agitating electro-magnetically magnetic beads in said sample preparation chamber.

66. ) The method of claim 63 further comprising heating said elution buffer contained in said filtration chamber to generate a thermal-gradient induced convection flow, and causing more nucleic acid molecules to enter into solution.

67. ) The method of claim 66 further comprising a step of adding surfactant to said elution buffer contained in said sample preparation chamber.

68. ) The method of claim 42 wherein said test sample is at least one of homogenized, digested and filtered before injection into said sample introduction inlet.

69. ) An apparatus for sample fluid analysis comprising a substrate, said substrate comprising:

a sample preparation chamber and fluidically coupled chamber or inlet for receiving a flow-promoting fluid;

a test sample introduction inlet fluidically coupled to said sample preparation chamber;

at least one buffer introduction inlet fluidically coupled to at least one of said sample preparation chamber and chamber or inlet for receiving a flow-promoting fluid;

a first multipurpose channel fluidically coupled to said sample preparation chamber or chamber or inlet for receiving a flow-promoting fluid;

an assay station;

an arrangement of at least first and second multipurpose channels wherein said at least one assay station being situated in a position intermediate and in fluid communication between said first and second multipurpose channels;

at least one flow control element for controlling fluid flow from at least one of said sample preparation chamber and chamber or inlet for receiving a flow-promoting fluid to said assay station;

at least one assay station channel fluidically coupled to said assay station.

70. ) The apparatus of claim 69 wherein said fluid communication is via at least first and second assay station channels in communication with said first and second multipurpose channels.

71. ) The apparatus of claim 69 wherein said filtration chamber comprises a sintered glass block which seals at least a portion of said a sample preparation chamber.

72. ) The apparatus of claim 70 wherein an absorbent adheres to said sintered glass block for filtering white blood cells through said sintered glass block.

73. ) The apparatus of claim 70 wherein a sealing layer seals at least one said first multi purpose channel, assay station, and first assay station channel.

74. ) The apparatus of claim 69 further comprising a diaphragm which seals at least a portion of said a sample preparation chamber and a vibrating actuator for vibrating said diaphragm.

75. ) The apparatus of claim 69 wherein said substrate comprises at least one of (a) glass, (b) plastic, (c) elastomer, (d) a composite, (e) silicon and (f) metal.

76. ) The apparatus of claim 75 wherein said elastomer comprises poly-dimethylsiloxane.