Device and method for the detection of an analyte utilizing mesoscale flow systems

• US 6,551,841 B1
• Filed: 01/27/1999
• Issued: 04/22/2003
• Est. Priority Date: 05/01/1992
• Status: Expired due to Term

• Current Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Kricka, Larry J., Wilding, Peter, Zemel, Jay N.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/237,523
• Time in Patent Office: 1,546 Days
• Field of Search: 435/6, 435/7.2, 435/7.21, 435/287.1, 435/287.9, 435/288.4, 435/288.5, 435/288.7, 435/810, 436/164, 436/514, 436/518, 436/524, 436/527, 436/531, 436/533, 436/534, 436/805, 436/806, 436/807, 436/809, 422/55-58
• US Class Current: 436/518
• CPC Class Codes: B01F 25/41 Mixers of the fractal type
• CPC Class Codes: B01F 35/7182 with means for feeding the material with a fractal or tree-type distribution in a surface
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• 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 2200/146 Employing pressure sensors
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• 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 2300/0822 Slides
• 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 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 2300/0883 : Serpentine channels
• CPC Class Codes: B01L 2300/0887 : Laminated structure
• CPC Class Codes: B01L 2300/1827 : using resistive heater
• CPC Class Codes: B01L 2300/1861 : using radiation
• CPC Class Codes: B01L 2400/0406 : capillary forces
• CPC Class Codes: B01L 2400/043 : magnetic forces
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/086 : using baffles or other fixed flow obstructions
• 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 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 3/50273 : characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502746 : characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502761 : specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• 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: B01L 7/525 : with physical movement of samples between temperature zones
• CPC Class Codes: B01L 9/527 : for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: C12Q 1/686 : Polymerase chain reaction [PCR]
• CPC Class Codes: C12Q 2565/629 : being a microfluidic device
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: Y10S 366/03 : Micromixers: variable geometry from the pathway influences mixing/agitation of non-laminar fluid flow
• CPC Class Codes: Y10S 435/81 : Packaged device or kit
• CPC Class Codes: Y10S 435/97 : Test strip or test slide
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/806 : Electrical property or magnetic property
• CPC Class Codes: Y10S 436/807 : Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 : Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 977/923 : Cell culture
• CPC Class Codes: Y10S 977/953 : Detector using nanostructure

Abstract

Disclosed are devices for detecting the presence of a preselected analyte in a fluid sample. The devices comprise a substrate microfabricated to define a sample inlet port, and a mesoscale flow system that includes a sample flow channel extending from the inlet port. The mesoscale flow system further includes an analyte detection region in fluid communication with the flow channel comprised of a binding moiety for specifically binding the analyte. The detection region is constructed with a mesoscale dimension sufficiently small to enhance binding of the binding moiety and the analyte. The binding moiety may be immobilized in the detection region. The mesoscale detection systems of the invention may be used in a wide range of applications, including the detection of cells or macromolecules, or for monitoring reactions or cell culture growth.

Claims

1. A device comprising a first polymer substrate layer having a plurality of mesoscale channels fabricated thereon, which first polymer substrate layer is overlaid by a flat cover layer, which cover layer comprises a transparent portion, whereby the plurality of channels are sealed between the first polymer substrate layer and the flat cover layer.

2. The device of claim 1, wherein the flat cover layer and the first substrate Isyer are adhered.

3. The device of claim 1, wherein the first polymer substrate is formed by molding the polymeric substrate.

4. The device of claim 1, wherein the cover layer comprises glass.

5. The device of claim 1, wherein the cover layer comprises a material other than glass.

6. The device of claim 1, wherein the cover layer comprises a plastic.

7. The device of claim 1, wherein the cover layer comprises a plastic sheet.

8. The device of claim 1, wherein the flow channels are fabricated on the first substrate by at least one of spin coating and vapor deposition, photolitography, wet chemical etching and plasma processing.

9. The device of claim 1, further comprising a second cover layer bonded to the first substrate.

10. The device of claim 1, wherein the plurality of channels intersect at a plurality of intersections.

11. The device of claim 1, wherein the channels are between about 0.5 and 500 microns in at least one channel dimension.

12. The device of claim 1, wherein the channels are between about 2 and 500 microns wide and between 0.1 and 500 microns deep.

13. The device of claim 1 further comprising a sample inlet port coupled to at least one of the plurality of channels.

14. The device of claim 1 further comprising at least one source of at least one biological material.

15. The device of claim 14, wherein the biological material comprises at least one of blood, plasma, serum, urine, sputum, saliva, cells and antibodies.

16. The device of claim 14, wherein the at least one source of at least one biological fluid is connected to a sample inlet port, which sample inlet port is coupled to at least one of the plurality of channels.

17. The device of claim 1, further comprising an appliance, the appliance comprising a nesting site for receiving the first substrate when overlayed by the transparent cover layer.

18. The device of claim 17, the appliance comprising a conduit for delivering source of sample material to at least one of the plurality of channels.

19. The device of claim 17, the appliance comprising an injector for injecting a sample into contact with at least one of the plurality of channels.

20. The device of claim 19, wherein the injector is a pressure injector and the appliance further comprises a pressure detector.

21. The device of claim 17, wherein the cover layer comprises a sample inlet port fluidly coupled to at least one of the plurality of channels, the appliance comprising a flow line coupled to the sample inlet port.

22. The device of claim 17, further comprising one or more valves in the appliance or in contact with one or more of the plurality of channels, which one or more valves regulates fluid flow.

23. The device of claim 1 further comprising means for sample movement.

24. The device of claim 1 further comprising means for reagent movement.

25. The device of claim 1 further comprising a detection region within at least one of the channels.

26. The device of claim 1 further comprising a conductivity sensor in at least one of the channels.

27. The device of claim 1 further comprising a pressure sensor in at least one of the channels.

28. The device of claim 1 further comprising an optical detector proximal to channel for detecting an optically detectable moiety within the channel.

29. The device of claim 28, wherein the optical detector comprises a spectroscope.

30. The device of claim 28, wherein the optical detector comprises a microscope.

31. The device of claim 28, wherein the optical detector comprises a light source.

32. The device of claim 28, wherein the optical detector comprises a camera.

33. The device of claim 28, wherein the optical detector detects a fluorescent or luminescent signal in a detection region coupled to or within a channel of the device.

34. The device of claim 1, further comprising a tilting mechanism for tilting the first substrate when overlaid by the cover layer.

35. The device of claim 1 further comprising a detection region within at least one mesoscale channel or chamber of the device, the detection region comprising a polymer bead disposed therein.

36. The device of claim 1 further comprising a plurality of electrical contacts configured to resistively heat a portion of the first substrate.

37. The device of claim 1 further comprising temperature control means for controlling the temperature of a portion of the device.

38. The device of claim 1 further comprising an inlet port and a source of a plurality of sample materials, which, during operation of the device, are flowed through the inlet port and into at least one of the plurality of channels.

39. The device of claim 1, the device further comprising an inlet port and a plurality of source of separate sample materials, which, during operation of the device, are flowed through the inlet port and into at least one of the plurality of channels.

40. The device of claim 1, wherein the first substrate is a few millimeters thick.

41. The device of claim 1, wherein the first substrate and cover layer are each between 0.2 and 2 centimeters square.

42. The device of claim 1, wherein the first substrate is manufactured from a polytetrafluoroethylene.

43. The device of claim 1, wherein the total volume of the plurality of channels is less than 10 microliters.

44. The device of claim 1, wherein the channels comprise at least one surface coating.

45. The device of claim 1, further comprising a magnetic bead disposed within one or more of the plurality of channels.

46. The device of claim 45, further comprising a magnetic field source of directing movement of the magnetic bead.

47. The device of claim 1, wherein at least one of the channels varies in width along the length of the at least one channel.

48. The device of claim 1, further comprising a pump coupled to a port coupled to at least one of the plurality of channels.

49. The device of claim 1, further comprising reagents for PCR amplification within the channels of the device.

50. The device of claim 1, further comprising a microprocessor coupled to a detector which is mounted proximal to the cover layer.

51. The device of claim 1, wherein the plurality of channels comprise at least one sample and at least one control region.

52. The device of claim 1, further comprising a microprocessor coupled to a detector which is mounted proximal at least one sample and at least one control region, which microprocessor compares data from the sample and the control region.