High throughput screening assay systems in microscale fluidic devices

• US 6,274,337 B1
• Filed: 03/19/1998
• Issued: 08/14/2001
• Est. Priority Date: 06/28/1996
• Status: Expired due to Term

• Current Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Original Assignee: Caliper Technologies Corp (Revvity, Inc.)
• Inventors: Parce, J. Wallace, Kopf-Sill, Anne R., Bousse, Luc J.
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): Pham, Minh-Quan K.
• Application Number:
  US09/044,557
• Time in Patent Office: 1,244 Days
• Field of Search: 435/5, 435/7.2, 435/7.21, 435/7.22, 435/7.23, 435/7.24, 435/7.25, 435/7.32, 435/7.31, 435/7.3, 435/29, 435/287.1, 435/287.2, 435/287.3, 435/288.5, 436/514, 436/536
• US Class Current: 435/29
• 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 2219/00828 Silicon wafers or plates
• CPC Class Codes: B01J 2219/00831 Glass
• CPC Class Codes: B01J 2219/00833 Plastic
• CPC Class Codes: B01J 2219/00853 Employing electrode arrangements
• CPC Class Codes: B01J 2219/00952 Sensing operations
• CPC Class Codes: B01J 2219/00961 Temperature
• CPC Class Codes: B01J 2219/0097 Optical sensors
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• 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 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 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• 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 2400/0418 : electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0487 : fluid pressure, pneumatics
• CPC Class Codes: B01L 3/5025 : for parallel transport of multiple samples
• 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 3/502753 : 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: 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 33/5064 : Endothelial cells
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/5097 : involving plant cells immunoassays of plant cells G01N33/56961
• 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 435/817 : Enzyme or microbe electrode
• CPC Class Codes: Y10S 436/804 : Radioisotope, e.g. radioimmunoassay
• CPC Class Codes: Y10S 436/805 : Optical property
• CPC Class Codes: Y10S 436/806 : Electrical property or magnetic property
• CPC Class Codes: Y10T 436/11 : Automated chemical analysis
• CPC Class Codes: Y10T 436/110833 : Utilizing a moving indicator strip or tape
• CPC Class Codes: Y10T 436/113332 : with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/117497 : with a continuously flowing sample or carrier stream
• CPC Class Codes: Y10T 436/25 : including sample preparation

Abstract

The present invention provides novel 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 a plurality of test compounds for an effect on a cell comprising:

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

m;

flowing a first biological cell in a first channel;

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

contacting the first biological cell in the first channel with the at least first test compound and,

detecting an effect of the at least first test compound on the first biological cell.

2. The method of claim 1, wherein the biological cell is isolated from an in vitro source.

3. The method of claim 1, wherein the biological cell is isolated from an in vivo source.

4. The method of claim 1, wherein the biological cell is selected from the group consisting of a bacterial cell, a plant cell, a fungal cell, and an animal cell.

5. The method of claim 1, wherein the biological cell is derived from a tissue homogenate or a blood sample.

6. The method of claim 1, wherein the biological cell is derived from a pathogen or tumor cell.

7. The method of claim 1, wherein the biological cell is infected with a virus.

8. The method of claim 1, wherein the biological cell is infected with HIV.

9. The method of claim 1, wherein the biological cell comprises one or more receptor molecules selected from the group consisting of transferrin, c-kit, a viral receptor, a cytokine receptor, a chemokine receptor, an interleukin receptor, an immunoglobulin receptor, an antibody, a cadherein family receptor, an integrin family receptor, and, a selectin family receptor.

10. The method of claim 1, wherein the biological cell comprises an intracellular receptor selected from the group consisting of a steroid receptor, a thyroid hormone receptor, a retinoid receptor and a vitamin D receptor.

11. The method of claim 1, wherein the test compound is selected from the group consisting of:

a toxin, a venom, a viral epitope, and a hormone.

12. The method of claim 1, wherein the cell comprises an indicator compound which interacts with the test compound to produce a detectable signal representative of a function of the cell.

13. The method of claim 1, wherein the biological cell comprises an enzyme and a substrate for the enzyme, wherein action of the enzyme on the substrate produces a detectable signal in the cell.

14. The method of claim 1, wherein the cell comprises a receptor/ligand binding pair, wherein at least one of the receptor or ligand has a detectable signal associated therewith.

15. The method of claim 1, wherein the cell comprises a receptor/ligand binding pair, wherein binding of the receptor to the ligand produces a detectable signal.

16. The method of claim 1, wherein the effect of the test compound is an ability of the test compound to enter the cell.

17. The method of claim 16, wherein the biological cell is an epithelial or endothelial cell.

18. The method of claim 1, wherein the effect is selected from the group consisting of cell proliferation, cell differentiation, cell activation, activation of a cell activity mediating enzyme, stimulation of messenger turnover in the cell, alteration of cell ion fluxes, activation of cellular enzymes, changes in cell shape, and an alteration in expression of a gene.

19. The method of claim 1, wherein the biological cell is capable of producing a detectable signal corresponding to a cellular function, and the detecting step comprises detecting an effect of the test compound on the cellular function by detecting a level of the detectable signal.

20. The method of claim 1, wherein the detecting step comprises detecting an effect of the test compound on viability of the cell.

21. The method of claim 1, wherein a pressure based fluid direction system is used for flowing said biological cell within said first channel and for flowing said test compound from said second channel into said first channel.

22. The method of claim 1, wherein an electrokinetic fluid direction system is used for flowing said biological cell within said first channel and for flowing said test compound from said second channel into said first channel.

23. A method of determining whether a sample contains a compound capable of affecting a cell, comprising:

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

m;

flowing a first cell in a first channel;

flowing the sample from a second channel into the first channel;

contacting the first cell in the first channel with the sample; and

,

detecting an effect of the sample on the cell.

24. The method of claim 23, wherein the first cell is isolated from an in vitro source.

25. The method of claim 23, wherein the first cell is isolated from an in vivo source.

26. The method of claim 23, wherein the first cell is selected from the group of cells consisting of a bacterial cell, a plant cell, a fungal cell, and an animal cell.

27. The method of claim 23, wherein the first cell is derived from a tissue homogenate or blood sample.

28. The method of claim 23, wherein the first cell is derived from a pathogen or tumor cell.

29. The method of claim 23, wherein the first cell is infected with a virus.

30. The method of claim 23, wherein the first cell is infected with HIV.

31. The method of claim 23, wherein the effect is selected from the group consisting of cell proliferation, cell differentiation, cell activation, activation of a cell activity mediating enzyme, stimulation of messenger turnover in the cell, alteration of cell ion fluxes, activation of cellular enzymes, changes in cell shape, and an alteration in expression of a gene.

32. The method of claim 23, wherein the first cell comprises one or more receptor molecules selected from the group consisting of transferrin, c-kit, a viral receptor ligand, a cytokine receptor, a chemokine receptor, an interleukin receptor, an immunoglobulin receptor;

an antibody, a cadherein family receptor, an integrin family receptor, and, a selectin family receptor.

33. The method of claim 23, wherein the at least first component of a biochemical system comprises at least one member of an antibody/antigen binding pair, wherein the antibody is specifically immunoreactive with the antigen.

34. The method of claim 23, wherein the cell comprises an antigen specifically reactive with an antibody.

35. The method of claim 23, wherein the sample comprises a test compound selected from the group consisting of:

a toxin, a venom, a viral epitope, and a hormone.

36. The method of claim 23, wherein the first cell comprises at least one member of a receptor/ligand binding pair.

37. The method of claim 23, wherein the first cell comprises a receptor and a ligand capable of specifically binding to the ligand.

38. The method of claim 23, wherein the first cell is derived from a patient.

39. The method of claim 23, wherein the detecting step comprises measuring a parameter of the cell in the presence and absence of the sample, and comparing the measured parameter in the presence of the sample to the measured parameter in the absence of the sample, a change in the parameter being indicative that the sample has an effect on the cell.

40. The method of claim 23, wherein a pressure based fluid direction system is used for flowing said first cell within said first channel and for flowing said sample from said second channel into said first channel.

41. The method of claim 23, wherein an electrokinetic fluid direction system is used for flowing said first cell within said first channel and for flowing said sample from said second channel into said first channel.