Analysis based on flow restriction

• US 5,486,335 A
• Filed: 04/24/1995
• Issued: 01/23/1996
• 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
• Primary Examiner(s): Warden, Robert J.
• Assistant Examiner(s): TRAN, HIEN THI
• Application Number:
  US08/427,493
• Time in Patent Office: 274 Days
• Field of Search: 422/55, 422/58, 422/61, 422/68.1, 422/73, 435/2, 435/7.2, 435/7.21, 435/6, 435/291, 436/164, 436/180, 436/501, 436/524, 436/809, 356/246
• US Class Current: 422/400
• 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 19/0046 Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays synthesis methods per se C40B50/00
• 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 2200/10 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 2300/0645 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 2300/0864 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 2400/0487 fluid pressure, pneumatics
• 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: 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: 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 436/809 : Multifield plates or multicontainer arrays

Abstract

Disclosed are devices and methods for detecting the presence of a preselected analyte in a fluid sample. The invention provides a device comprising a solid substrate, typically on the order of a few millimeters thick and approximately a 0.2 to 2.0 centimeters square, microfabricated to define a sample inlet port and a mesoscale flow system. A sample is passed through the mesoscale flow system, and the restriction or blockage of flow through the flow system is detected as a positive indication of the presence of the analyte. The mesoscale flow system includes in one embodiment a primary sample flow channel extending from the inlet port, and a fractal region, in fluid communication with the flow channel, comprising bifurcations leading to plural secondary flow channels. The device may be adapted for operation in conjunction with a pump, for example, to induce flow of a sample through the flow system. A detector may also be provided for detecting analyte induced changes in flow properties in the mesoscale flow system. The devices of the invention may be used in a wide range of applications, including the detection of cells or macromolecules, such as viruses.

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 primary sample flow channel extending from said inlet port; and

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

means for detecting a flow property of a fluid sample in said flow system as an indication of the presence of an analyte in the fluid sample.

2. The device of claim 1 wherein said fractal region further comprises junctions in fluid communication with said secondary flow channels leading to a third flow channel in said mesoscale flow system.

3. The device of claim 1 further comprising means for inducing flow of said sample through said mesoscale flow system.

4. The device of claim 2 wherein said fractal region comprises equal numbers of bifurcations and junctions disposed serially along the direction of flow of the sample through said fractal region.

5. The device of claim 4, wherein the fractal region has a reduced cross-sectional area relative to said primary flow channel and said third flow channel.

6. The device of claim 3, wherein said means for detecting comprises means for detecting analyte induced restriction of flow through said flow system.

7. The device of claim 2 wherein said means for detecting comprises means for detecting a parameter in said third flow channel.

8. The device of claim 2, wherein said means for detecting comprises means for detecting and comparing a parameter in said primary sample flow channel with a parameter in said third flow channel.

9. The device of claim 7 or 8 wherein said means for detecting comprises an electrical detection means.

10. The device of claim 7 wherein said parameter is fluid pressure.

11. The device of claim 7 wherein said parameter is fluid conductivity.

12. The device of claim, 1 wherein said means for detecting comprises means for detecting a parameter in said fractal region.

13. The device of claim 12 wherein said detecting means comprises means defining an optical path to said fractal region.

14. The device claim 1 further comprising a binding moiety disposed within said fractal region for binding the analyte of said sample.

15. The device of claim 14 wherein said binding moiety comprises particles which bind with the analyte of said sample to induce particle agglomeration.

16. The device of claim 1 wherein said means for detecting comprises a magnetic detection means.

17. The device of claim 1 wherein said substrate defines a plurality of said flow systems.

18. The device of claim 1 wherein said means for detecting comprises means for detecting the growth of an organism in said flow system.

19. The device of claim 1 wherein the sample is a sperm sample and wherein flow of sperm through the fractal region provides an indication of sperm motility.

20. The device of claim 19, wherein a channel in said substrate is disposed at an angle with respect to a horizontal plane.

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

22. 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 the inlet port on said substrate; and

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

23. The device of claim 1, wherein said means for detecting comprises 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.

24. The device of claim 23, 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.

25. The device of claim 1 wherein said flow system includes a control fractal region permitting comparison of flow of said sample in said fractal region and said control fractal region.

42. The device of claim 1, 4 or 25 wherein, within at least a portion of a channel in said flow system, each of the channel width and channel depth is between 0.1 μ

m and 500 μ

m.

43. The device of claim 42 wherein the channel width in said portion is between 2.0 and 300 μ

m.

44. The device of claim 42 wherein the channel depth in said portion is between 0.1 and 100 μ

m.

26. A method for detecting the presence or absence 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 primary sample flow channel extending from said inlet port; and

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

(ii) passing a fluid sample suspected to contain an analyte through said mesoscale flow system;

(iii) detecting the restriction or blockage of flow of the fluid sample through said system; and

(iv) correlating the detected restriction or blockage of flow, or lack of said detected restriction or blockage of flow, to the presence or absence of an analyte in said sample.

27. The method of claim 26, wherein said flow system comprises a binding moiety, capable of binding said analyte in said sample, to promote said restriction or blockage of flow through said system.

28. The method of claim 27 wherein said binding moiety is disposed on particles which bind with said analyte in said sample to induce particle agglomeration, thereby to promote said restriction or blockage of flow.

29. The method of claim 27 wherein said analyte is a cell population in said sample;

said binding moiety comprises a crosslinker of cells in said population; and

said restriction of flow is caused by crosslinker-induced cell aggregation.

30. The method of claim 27 wherein said binding moiety is immobilized within said flow system.

31. The method of claim 30 wherein flow is restricted by the build-up of a macromolecular surface layer on a surface of said flow system.

32. The method of claim 26 wherein said fractal region further comprises junctions in fluid communication with said secondary flow channels leading to a third channel in said flow system, to amplify the effect of occlusion of said flow system or increase in viscosity within said substrate.

33. The method of claim 26 wherein, in step (iii), restriction or blockage is detected electrically.

34. The method of claim 26 wherein, in step (iii), restriction or blockage is detected optically.

35. The method of claim 32 wherein said fractal region further comprises a binding moiety for binding the analyte of said sample.

36. The method of claim 35 wherein said binding moiety contains particles which bind with the analyte of said sample to induce particle agglomeration.

37. The method of claim 26, wherein said substrate, provided in step (i), further comprises a control region in fluid communication with said sample inlet port;

and

wherein, in step (iii), flow of said sample in said fractal region and said control region is detected and compared.

38. The method of claim 26 wherein the analyte comprises a replicable procaryotic organism;

and

wherein, in step (iii), the restriction or blockage of flow of said organism through said flow system serves as an indication of the presence of said organism.

41. The method of claim 32 or 40 wherein said fractal region comprises equal numbers of bifurcations and junctions disposed serially along the direction of flow of the sample through said fractal region.

45. The method of claim 26, 39 or 40 wherein, within at least a portion of a channel in said flow system, the channel width and channel depth each are between 0.1 μ

m and 500 μ

m.

46. The method of claim 45 wherein the channel width in said portion is between 2.0 and 300 μ

m.

47. The method of claim 45 wherein the channel depth in said portion is between 0.1 and 100 μ

m.

39. 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 primary sample flow channel extending from said inlet port; and

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

(ii) passing a fluid sample suspected to contain an analyte through said mesoscale flow system;

(iii) detecting a flow property of the fluid sample in the flow system; and

(iv) correlating the detected flow property of the fluid sample to the presence or absence of an analyte in the sample.

40. The method of claim 39 wherein said fractal region further comprises junctions, in fluid communication with the secondary flow channels, leading to a third flow channel.