Mesoscale detection structures

• US 20030129671A1
• Filed: 01/21/2003
• Published: 07/10/2003
• Est. Priority Date: 05/01/1992
• Status: Active Grant

• Current Assignee: Jay N. Zemel, Larry J. Kricka, Peter Wilding
• Original Assignee: Jay N. Zemel, Larry J. Kricka, Peter Wilding
• Inventors: Kricka, Larry J., Wilding, Peter, Zemel, Jay N.
• Granted Patent: US 7,018,830 B2
• Time in Patent Office: Days
• US Class Current: 435/7.2
• 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 for detecting the presence of an analyte in a fluid sample, the device comprising a solid substrate microfabricated to define:

a sample inlet port; and

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

an analyte detection region in fluid communication with said flow channel comprising a binding moiety for specifically binding said analyte, said detection region having a mesoscale dimension.

2. The device of claim 1 wherein said binding moiety is immobilized in said detection region.

3. The device of claim 1, wherein the device further includes a window for optically probing said detection region disposed over said region on said substrate.

4. The device of claim 1 wherein said substrate further defines means for delivering to said detection region a labeled substance which binds to said analyte to produce a detectable signal indicative of the presence of said analyte.

5. The device of claim 4 further comprising:

a window for optically probing said detection region disposed over said region on said substrate; and

optical means for detecting the presence of said detectable signal through said window.

6. The device of claim 1 wherein said analyte is an antigen and said binding moiety is an antigen binding protein.

7. The device of claim 1 wherein said analyte is a polynucleotide and said binding moiety is a complementary polynucleotide which hybridizes with said polynucleotide.

8. The device of claim 1 wherein said analyte and said immobilized binding moiety comprise a ligand/receptor pair.

9. The device of claim 4 wherein said optically detectable signal is a luminescent signal.

10. The device of claim 9 wherein said optically detectable signal is a fluorescent signal.

11. The device of claim 3 wherein said analyte detection region comprises particles having analyte binding sites on the surface thereof which, in the presence of an analyte induce particle agglomeration optically detectable through said window.

12. The device of claim 3 wherein said substrate further defines a control region in fluid communication with said sample inlet port and a control region window, disposed over said control region on said substrate, for optically probing said control region whereby data determined optically in said control and detection regions may be compared.

13. The device of claim 3 wherein said analyte is a cell population in said sample, and wherein

said binding moiety binds to a surface protein on members of said cell population to induce agglutination of said cell population;

and

said agglutination is optically detectable through said window.

14. The device of claim 1 wherein said substrate defines at least a second mesoscale flow system.

15. The device of claim 14 wherein the analyte detection regions of said flow systems comprise different immobilized binding moieties.

16. The device of claim 1, wherein the analyte is an intracellular molecular component in a cell-containing liquid biological sample, the device further comprising:

cell lysing means in said mesoscale flow system in fluid communication with said flow channel; and

means for forcing cells in said cell-containing microsample within said substrate into said cell lysing means thereby to release said intracellular molecular component.

17. The device of claim 16 wherein said cell lysing means comprises a portion of said flow channel having cell-membrane piercing protrusions extending from a wall thereof.

18. The device of claim 16 wherein said cell lysing means comprises a region of restricted cross-sectional dimension sufficient to permit passage of intracellular molecules while prohibiting passage of cells.

19. The device of claim 1, wherein said analyte is a cell population in said sample, said device further comprising:

a cell separation region comprising sites for binding cell surface protein immobilized on a wall of said separation region; and

means for inducing flow of said microsample to said separation region;

at a first flow rate sufficiently slow to permit capture of said cell population in the sample by said binding sites to separate said cell population from said microsample; and

at a second, higher, flow rate sufficient to release said separated cell fraction from said region.

20. The device of claim 3 wherein said flow system further comprises:

a fractal region in fluid communication with said flow channel comprising bifurcations leading to plural secondary flow channels; and

means for inducing flow of a biological sample through said flow channel and said fractal region.

21. The device of claim 1 wherein said solid substrate comprises microfabricated silicon.

22. The device of claim 1 wherein said sample flow channel and said detection region are microfabricated in a surface of a solid substrate and enclosed by a cover adhered to said surface.

23. The device of claim 1 further comprising an appliance for use in combination with said substrate, said appliance comprising:

means for holding said substrate,

fluid input means interfitting with an inlet port on said substrate, and

pump means for passing fluid through the flow system of said substrate when it is held in said holding means.

24. The device of claim 23 wherein said appliance further comprises a reagent reservoir and means for delivering a reagent to said flow system.

25. The device of claim 1, further comprising an appliance for use in combination with said substrate, said appliance comprising:

means for holding said substrate; and

optical means for viewing the contents of said mesoscale flow system in said substrate.

26. The device of claim 25, wherein said optical means comprises magnifying optics and a video camera, and wherein said appliance further comprises:

a tilt mechanism for manually adjusting the angle and location of the device; and

a video screen for viewing the contents of said flow system.

41. The device of claim 1, wherein said analyte detection region comprises a tortuous mesoscale flow channel;

and

wherein said tortuous channel is microfabricated with a length allowing the timed mixing and reaction of fluid flowing through said tortuous channel.

27. A method for detecting the presence of an analyte in a fluid sample, the method comprising the steps of:

(i) providing a device comprising a solid substrate microfabricated to define;

a sample inlet port; and

a mesoscale flow system comprising;

a sample flow channel extending from said inlet port; and

an analyte detection region in fluid communication with said flow channel comprising a binding moiety for specifically binding said analyte, said detection region having a mesoscale dimension;

(ii) delivering said sample to said inlet port and through said flow system to said detection region; and

(iii) detecting the binding of said analyte to said binding moiety in said detection region.

28. The method of claim 27 wherein said binding moiety in the device provided in step (i) is immobilized in said detection region;

and

wherein step (iii) includes the step of detecting the binding of said analyte and said immobilized binding moiety.

29. The method of claim 27, wherein the device provided in step (i) further includes a window disposed over said detection region on said substrate;

and

wherein step (iii) includes the step of optically probing said detection region through said window.

30. The method of claim 27 wherein the device provided in step (i) further includes means for delivering to said detection region a labeled substance which binds to said analyte to produce a detectable signal indicative of the presence of said analyte, the method further comprising:

(iv) delivering a labeled substance to said detection region; and

(v) optically probing for said detectable signal in said detection region.

31. The method of claim 30 wherein said optical probing step includes the step of detecting a luminescent signal.

32. The method of claim 27 wherein said analyte detection region, in the device provided in step (i), comprises particles having analyte binding sites on the surface thereof which, in the presence of an analyte, induce particle agglomeration;

wherein, in step (ii), said analyte binds to said particles to induce agglomeration; and

wherein, in step (iii), agglomeration is detected.

33. The method of claim 29 wherein said substrate provided in step (i) further defines a control region in fluid communication with said sample inlet port and a control region window, disposed over said control region on said substrate, for optically probing said control region whereby data determined optically in said control and detection regions may be compared;

and

wherein step (iii) includes the step of optically probing and comparing said control region and said detection region.

34. The method of claim 29 wherein said analyte is a cell population in said sample, and wherein,

in step (ii), said binding moiety binds to a surface protein on members of said cell population to induce agglutination of said cell population;

and

in step (iii), said agglutination is optically detected through said window.

35. The method of claim 27 wherein said substrate, provided in step (i), defines at least a second said mesoscale flow system;

and

wherein, in step (iii), binding in the detection regions in said first and at least second system is detected.

36. The method of claim 27, wherein the analyte is an intracellular molecular component in a cell-containing liquid biological sample, said method comprising the additional step of lysing said cells within said substrate to release said intracellular analyte prior to step (iii).

37. The method of claim 27, wherein said analyte is a cell population in said sample, said method comprising the additional step of separating said cell population from other cells within said substrate prior to step (iii).

42. The method of claim 27, wherein said analyte detection region further comprises a tortuous mesoscale flow channel, upstream from said detection reaction;

wherein said tortuous channel is microfabricated with a length allowing the timed mixing and reaction of fluid flowing through said tortuous channel prior to delivery to said detection region; and

wherein step (ii) includes the step of delivering said sample to said tortuous channel then to said detection region.

38. An appliance for use with an analytical device including a solid substrate microfabricated to define at least one inlet port and a mesoscale flow system, extending from an inlet port on said substrate, the appliance comprising:

means for holding said substrate;

fluid input means interfitting with an inlet port on said substrate; and

pump means for passing fluid through the flow system of said substrate when is held in said holding means.

39. The appliance of claim 38, further comprising a reagent reservoir and means for delivering a reagent to said flow system.

40. The appliance of claim 39, further comprising detection means for detecting a parameter of a fluid sample in said flow system.