High throughput screening assay systems in microscale fluidic devices

• US 5,942,443 A
• Filed: 06/28/1996
• Issued: 08/24/1999
• Est. Priority Date: 06/28/1996
• Status: Expired due to Term

• Current Assignee: Caliper Life Sciences Incorporated (Revvity, Inc.)
• Original Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Inventors: Parce, John Wallace, Kopf-Sill, Anne R., Bousse, Luc J.
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): PORTNER, VIRGINIA ALLEN
• Application Number:
  US08/671,987
• Time in Patent Office: 1,152 Days
• Field of Search: 436/514, 435/4, 435/6
• US Class Current: 506/39
• 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: 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 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/0673 Handling of plugs of fluid surrounded by immiscible fluid
• CPC Class Codes: B01L 2200/0689 Sealing
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• 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 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0887 Laminated structure
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• 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 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• 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 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 9/527 for microfluidic devices, e.g. used for lab-on-a-chip
• CPC Class Codes: G01N 27/44791 Microapparatus sample containers with integrated microfluidic structures B01L3/5027
• CPC Class Codes: G01N 33/5008 : for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
• CPC Class Codes: G01N 33/5011 : for testing antineoplastic activity
• CPC Class Codes: G01N 33/502 : for testing non-proliferative effects
• CPC Class Codes: G01N 33/5044 : involving specific cell types
• CPC Class Codes: G01N 33/5064 : Endothelial cells
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5094 : for blood cell populations red blood cells G01N33/80
• CPC Class Codes: G01N 33/5304 : Reaction vessels, e.g. agglutination plates for solid-phase systems G01N33/543
• CPC Class Codes: G01N 33/57415 : of breast
• CPC Class Codes: G01N 35/08 : using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
• CPC Class Codes: Y10S 366/02 : Micromixers: segmented laminar flow with boundary mixing orthogonal to the direction of fluid propagation with or without geometry influences from the pathway
• 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

Abstract

Microfluidic devices and methods that are useful for performing high-throughput screening assays. In particular, the devices and methods of the invention are useful in screening large numbers of different compounds for their effects on a variety of chemical, and preferably, biochemical systems.

Claims

1. A method of screening test compounds for an effect on an interaction between components of a biochemical system, comprising;

providing a substrate having at least two intersecting channels disposed therein, at least one of said at least two intersecting channels having at least one cross-sectional dimension in a range from 0.1 to 500 μ

m;

continuously flowing interacting components of a biochemical system in a first of said at least two intersecting channels;

flowing at least a first test compound from a second channel into said first channel whereby said first flowing test compound contacts said flowing components of said biochemical system; and

detecting an effect of said at least first flowing test compound on interactions between said continuously flowing components of said biochemical system.

2. The method of claim 1, wherein said components of a biochemical system produce a detectable signal representative of a function of said biochemical system.

3. The method of claim 1, wherein said components further comprise an indicator compound which interacts with said components to produce a detectable signal representative of a functioning of said biochemical system.

4. The method of claim 1, wherein components of a biochemical system comprise an enzyme and a substrate for said enzyme, wherein action of said enzyme on said substrate produces a detectable signal.

5. The method of claim 1, wherein said components of a biochemical system comprise a receptor/ligand binding pair, wherein at least one of said receptor or ligand has a detectable signal associated therewith.

6. The method of claim 1, wherein said components of a biochemical system comprise a receptor/ligand binding pair, wherein binding of said receptor to said ligand produces a detectable signal.

7. The method of claim 1, wherein said components of a biochemical system comprise components of cells, which cells are flowing in said first channel, and said detecting step comprises determining an effect of said test compound on said cells.

8. The method of claim 7, wherein said cells are capable of producing a detectable signal corresponding to a cellular function, and said detecting step comprises detecting an effect of said test compound on said cellular function by detecting a level of said detectable signal.

9. The method of claim 7, wherein said detecting step comprises detecting an effect of said test compound on viability of said cells.

35. The method of claim 1, wherein said components and said first test compound are flowed electroosmotically.

36. The method of claim 1, wherein said electrokinetic fluid direction system is used for flowing said components within said first channel and for introducing said test compound from said second channel to said first channel.

37. The method of claim 36, further comprising generating a continuous flow of a fluid mixture comprising said components, and periodically injecting the first test compound from said second channel in to the continuous flow of the fluid mixture in said first channels.

38. The method of claim 1 wherein said components and said test compound are flowed by concomitantly applying a variable voltage to at least three electrodes in electrical contact with said at least two intersecting channels.

10. A method of screening a plurality of test compounds for an effect on a biochemical system, comprising:

providing a substrate having at least two intersecting channels disposed therein, at least one of said at least two intersecting channels having at least one cross-sectional dimension in a range from 0.1 to 500 μ

m;

continuously flowing interacting components of a biochemical system in a first channel of said at least two intersecting channels;

periodically introducing a different flowing test compound into said first channel from a second channel of said at least two intersecting channels; and

detecting an effect of said test compound on said continuously flowing interacting components of a biochemical system.

11. The method of claim 10, wherein said step of periodically introducing comprises flowing a plurality of different test compounds into said first channel from a second channel of said at least two intersecting channels, each of said plurality of different test compounds being physically isolated from each other of said plurality of different test compounds.

12. The method of claim 10, wherein said components of a biochemical system produce a detectable signal representative of a function of said biochemical system.

13. The method of claim 12, wherein said detecting comprises monitoring said detectable signal from said continuously flowing components at a point on said first channel, said detectable signal having a steady state intensity, and wherein said effect of said interaction between said components and said test compound comprises a deviation from said steady state intensity of said detectable signal.

14. The method of claim 12, wherein said components of a biochemical system comprise an indicator compound which interacts with at least a first component of a biochemical system to produce a detectable signal representative of a functioning of said biochemical system.

15. The method of claim 14, wherein said components of a biochemical system comprises an enzyme and said indicator compound comprises a substrate for said enzyme, wherein action of said enzyme on said substrate produces a detectable signal.

16. The method of claim 12, wherein said components of a biochemical system comprise a receptor/ligand binding pair, wherein at least one of said receptor or ligand has a detectable signal associated therewith.

17. The method of claim 16, wherein said receptor and said ligand flow along said first channel at different rates.

18. The method of claim 12, wherein said components of a biochemical system comprises a receptor/ligand binding pair, wherein binding of said receptor to said ligand produces a detectable signal.

19. The method of claim 10, wherein said components of a biochemical system comprise cells, and said detecting step comprises determining an effect of said test compound on said cells.

20. The method of claim 19, wherein said cells are capable of producing a detectable signal corresponding to a cellular function, and said detecting step comprises detecting an effect of said test compound on said cellular function by detecting a level of said detectable signal.

21. The method of claim 19, wherein said detecting step comprise s detecting an effect of said test compound on viability of said cells.

39. The method of claim 10, wherein said components and said different test compounds are flowed electroosmotically.

40. The method of claim 10, wherein an electrokinetic fluid direction system is used for flowing said components within said first channel and for introducing said different test compound from said second channel to said first channel.

41. The method of claim 40, further comprising generating a continuous flow of a fluid mixture comprising said components, and periodically injecting the different test compound from said second channel into the continuous flow of the fluid mixture in said first channel.

42. The method of claim 10, wherein said components and said test compound are flowed by concomitantly applying a variable voltage to at least three electrodes in electrical contact with said at least two intersecting channels.

22. A method of screening a plurality of different test compounds for an effect on a biochemical system, comprising:

providing a substrate having at least a first surface, and a plurality of reaction channels fabricated in said first surface, each of said plurality of reaction channels being fluidly connected to at least two transverse channels fabricated in said surface;

introducing components of a biochemical system into said plurality of reaction channels;

flowing a plurality of different test compounds through at least one of said at least two transverse channels, each of said plurality of test compounds being introduced into said at least one transverse channels in a discrete volume;

directing each of said plurality of different test compounds into a separate one of said plurality of reaction channels; and

detecting an effect of each of said test compounds on said components of said biochemical system.

23. The method of claim 22, wherein said components of said biochemical system produce a flowable detectable signal representative of a function of said biochemical system.

24. The method of claim 23, wherein said detectable flowable signal produced in each of said plurality of reaction channels is flowed into and through said second transverse channel, each of said detectable flowable signals produced in each of said plurality of reaction channels being physically isolated from each other of said detectable flowable signals, whereupon each of said detectable flowable signals is separately detected.

25. The method of claim 23, wherein said flowable signal comprises a soluble signal.

26. The method of claim 25, wherein said soluble signal is selected from fluorescent or colorimetric signals.

27. The method of claim 22, wherein said components comprise an indicator compound which interacts with at least a first component of a biochemical system to produce a detectable signal representative of a functioning of said biochemical system.

28. The method of claim 27, wherein said first component of a biochemical system comprises an enzyme and said indicator compound comprises a substrate for said enzyme, wherein action of said enzyme on said substrate produces a detectable signal.

29. The method of claim 22, wherein said components of a biochemical system comprise a receptor/ligand binding pair, wherein at least one of said receptor or ligand has a detectable signal associated therewith.

30. The method of claim 22, wherein said components of a biochemical system comprise a receptor/ligand binding pair, wherein binding of said receptor to said ligand produces a detectable signal.

31. The method of claim 22, wherein said components of a biochemical system comprise cells, and said detecting step comprises determining an effect of said test compound on said cells.

32. The method of claim 31, wherein said cells are capable of producing a detectable signal corresponding to a cellular function, and said detecting step comprises detecting an effect of said test compound on said cellular function by detecting a level of said detectable signal.

33. The method of claim 32, wherein said detecting step comprises detecting an effect of said test compound on viability of said cells.

34. The method of claim 22, wherein each of said plurality of different test compounds is immobilized upon a separate bead, and said step of directing each of said plurality of different test compounds into a separate one of said plurality of reaction channels comprises:

lodging one of said separate beads at an intersection of said first transverse channel and each of said plurality of reaction channels; and

controllably releasing said test compounds from each of said separate beads into each of said plurality of reaction channels.

43. The method of claim 22, wherein said components and said plurality of different test compound are flowed electroosmotically.

44. The method of claim 22, wherein an electrokinetic fluid direction system is used for flowing said components within said first channel and for introducing said plurality of test compounds from said second channel to said first channel.

45. The method of claim 44, further comprising generating a continuous flow of a fluid mixture of said components along said first channel, and periodically injecting a one or more of the plurality of test compounds from said second channel into the fluid mixture in said first channel.

46. The method of claim 22, wherein said components and said plurality of test compounds are flowed by concomitantly applying a variable voltage to at least three electrodes in electrical contact with said at least two intersecting channels.

47. A method of screening a plurality of test compounds for an effect on a biochemical system, comprising:

providing a first channel and a second channel which intersect, wherein at least one of said first and second channels has a cross sectional dimension in the range from 0.1 to 500 μ

m;

continuously flowing interacting components of the biochemical system into said first channel;

introducing at least a first flowing test compound into the first channel from the second channel; and

detecting an effect of the first flowing test compound on an interaction between the continuously flowing components of the biochemical system.

48. The method of claim 47 further comprising providing a detection zone in said first or said second channel.

49. The method of claim 47, wherein the components comprise a component of a receptor/ligand binding pair.

50. The method of claim 47, wherein the components are selected from an enzyme, an enzyme substrate, and a biological cell.

51. The method of claim 47, wherein detecting the effect of the first test compound on the biochemical system comprises monitoring a fluorescent signal.

52. The method of claim 47, further comprising flowing a second component of the biochemical system into said first channel.

53. The method of claim 47, further comprising flowing a second test compound into said first channel.

54. The method of claim 47, further comprising flowing a second test compound into said first channel, wherein the first test compound and the second test compound are not mixed.

55. The method of claim 47, wherein said components and said first test compound are flowed electroosmotically.

56. The method of claim 47, wherein an electrokinetic fluid direction system is used for flowing said components within said first channel and for introducing said test compound from said second channel to said first channel.

57. The method of claim 56, further comprising generating a continuous flow of a fluid mixture comprising said components along the first channel, and periodically injecting a test compound from said second channel into the fluid mixture in said first channel.

58. The method of claim 47, wherein said components and said test compound are flowed by concomitantly applying a variable voltage to at least three electrodes in electrical contact with said at least two intersecting channels.

59. The method of claim 47, wherein the components comprise a biological cell and the method further comprises the step of detecting the effect of the first test compound on the cell.

60. The method of claim 47, wherein the components comprise an enzyme and an enzyme substrate and the method further comprises detecting the formation of a enzyme-substrate product.

61. The method of claim 47, wherein the components comprise a receptor and a ligand and the method further comprises detecting the binding of the ligand to the receptor.

62. The method of claim 47, wherein the components produce a detectable signal representative of a function of the biological system.

63. The method of claim 47, wherein the first and second intersecting channels are fabricated into a planar substrate.

64. A method of screening a plurality of test compounds for an affect on a biochemical system, comprising:

providing a substrate having at least one surface with a plurality of reaction channels fabricated into said surface;

providing at least two transverse channels fabricated into said surface, each of said plurality of reaction channels having fluidly connected to a first of said at least two transverse channels at a first point in said reaction channels and fluidly connected to a second of said at least two transverse channels at a second point in said reaction channels, and at least two transverse channels and said plurality of reaction channels each having at least one cross-sectional dimension in the range from about 0.1 to about 500 μ

m;

providing a biochemical source of components of said biochemical system, said source of components of said biochemical system being fluidity connected to one or more of said plurality of reaction channels;

providing a test source of test compounds fluidly connected to said first of said at least two transverse channels;

providing a fluid direction system for controlling movement of said test compound and said components within said at least two transverse channels and said plurality of reaction channels;

providing a cover mated with said surface;

continuously flowing components of the biochemical system from said biochemical source through at least one of said reaction channels;

flowing a first test compound from said test source through at least one of said reaction channels;

mixing said continuously flowing components of the biochemical system and said flowing test compound in at least one of said reaction channels; and

detecting an effect of said first test compound on the biochemical system.

65. The method of claim 64, wherein the flowing components of the biochemical system comprise a receptor/ligand binding pair, wherein at least one of said receptor or ligand has a detectable signal associated therewith.

66. The method of claim 64, wherein the flowing components of the biochemical system comprise an enzyme and an enzyme substrate and the method further comprises detection formation of a product produced by interaction between the enzyme and the substrate.

67. The method of claim 64, wherein the method further comprises detecting a fluorescent or chromogenic signal to quantify the effect of said first test compound on the biochemical system.

68. The method of claim 64, further comprising providing a detection zone in at least one of said plurality of transverse channels.

69. The method of claim 64, further comprising providing a detection system for detecting the effect of the test compound on the biochemical system.

70. The method of claim 64, further comprising providing a fluid direction system for controlling movement of said test compound and said components within said at least two transverse channels.

71. The method of claim 70, wherein the fluid direction system comprises a plurality of individual electrodes, each in electrical contact with each terminus of said at least two transverse channel;

and

a control system for concomitantly applying a variable voltage at each of said electrodes, whereby movement of said test compounds or said components in said at least two transverse channels and said plurality of reaction channels are controlled.