Pre-disposed assay components in microfluidic devices and methods

• US 6,479,299 B1
• Filed: 08/12/1998
• Issued: 11/12/2002
• Est. Priority Date: 06/28/1996
• Status: Expired due to Term

• Current Assignee: Caliper Holdings Incorporated
• Original Assignee: Caliper Holdings Incorporated
• Inventors: Parce, J. Wallace, Kopf-Sill, Anne R., Bousse, Luc J.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US09/132,963
• Time in Patent Office: 1,553 Days
• Field of Search: 204/601, 204/450, 204/451, 204/600, 436/29, 436/62, 436/149, 436/163, 436/806, 436/150, 436/52, 436/514, 422/68.1, 422/81, 422/82, 435/4
• US Class Current: 436/514
• 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

Microfluidic devices having predisposed assay components for increased throughput and prolonged shelf life are provided.

Claims

1. A method of manufacturing a microfluidic apparatus comprising:

providing a microfluidic device comprising a body having at least two enclosed, fluidly connected intersecting microfluidic channels disposed therein such that at least one of said at least two intersecting microfluidic channels is fluidly connected to a fluid reservoir; and

disposing an assay component into at least one of the at least two intersecting channels by delivering said assay component using a material transport system.

2. The method of claim 1 wherein the delivered assay component is in stabilized form.

3. The method of claim 2, wherein the assay component is stabilized for extended shelf-life.

4. The method of claim 1, further comprising a source of a plurality of different test compounds fluidly connected to a first of said at least two intersecting microfluidic channels.

5. The method of claim 1 wherein the material transport system comprises a fluid direction system for flowing said assay component to a second of said at least two intersecting microfluidic channels and introducing a plurality different test compounds form said first to said second of said at least two intersecting microfluidic channels.

6. The method of claim 1, wherein the assay component is lyophilized.

7. The method of claim 1, wherein the assay component is immobilized on a bead.

8. The method of claim 1, wherein at least a second assay component is predisposed in at least one of said at least two channels, or within said reservoir, wherein said second assay component is in stabilized form.

9. The method of claim 8, wherein the first and second assay components contact one another in an assay performed in at least one of said at least two intersecting microfluidic channels.

10. The method of claim 1, wherein the assay component is reconstituted by introducing a buffer solution into the reservoir or by flowing the buffer solution through one or more of the at least two channels.

11. The method of claim 1, wherein said assay component is a receptor-ligand interaction assay component, an enzyme-substrate interaction assay component, an antibody-antigen interaction assay component, a cellular signaling pathway assay component, or a transport reaction assay component.

12. The method of claim 1, wherein the assay component comprises an enzyme, a receptor-ligand binding pair, 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, a selectin family receptor, a steroid receptor, a thyroid hormone receptor, a retinoid receptor, a vitamin D receptor, a toxin, a venom, a viral epitope, a hormone, an anti-coagulant, a detectable ligand, a colorimetric ligand, or a fluorescent ligand.

13. The method of claim 1, wherein the assay component produces an effect in an assay, the effect selected from the group consisting of:

cell proliferation, cell differentiation, cell activation, activation of a cell activity mediating enzyme, stimulation or messenger turnover in a cell, alteration a cell ion fluxes, activation of cellular enzymes, a change in cell shape, and an alteration in expression of a gene.

14. The method of claim 1, wherein the assay component is blood-derived.

15. The method of claim 1, wherein one of said at least two intersecting microfluidic channels has a cross-sectional dimension in the range from about 0.1 to about 500 μ

m.

16. The method of claim 1, wherein one of said at least two intersecting microfluidic channels has a serpentine, square-wave, saw tooth, or reciprocating channel geometry.

17. The method of claim 1, wherein said body is fabricated by a process comprising a step selected from (i) molding a polymer, or (ii) by wet etching said at least two intersecting microfluidic channels on a glass substrate and thermally fusing a cover layer to the glass substrate.

18. The method of claim 1, wherein said body is fabricated from quartz, silicon, glass, or plastic.

19. The method of claim 1, wherein said body comprises a planar surface.

20. The method of claim 1, wherein the microfluidic device further comprises a plurality of electrodes in a configuration which mates with a plurality of reservoirs in the body, the reservoirs being fluidly connected to one or more of said intersecting microfluidic channels;

and, a control system for concomitantly applying a voltage to at least two of the plurality of electrodes, whereby movement of said first component in said at least two intersecting microfluidic channels is electrokinetically controlled.

21. The method of claim 1, further comprising an assay component detection system.

22. The method of claim 21, wherein said detection system includes a detection window in said second channel.

23. The method of claim 21, wherein said detection system comprises a fluorescent detector.

24. The method of claim 1, wherein said assay component is moved electrokinetically through at least one of said at least two intersecting microfluidic channels.

25. The method of claim 1, wherein the body is a multi-layered structure comprising at least a first and second planar substrate, which first and second planar substrates are bonded to form the body.

26. The method of claim 1, wherein the body comprises first and second bonded planar substrate layers,

wherein the first substrate layer comprises one or more of: a polymer, plastic, glass, quartz, and silicon,

and the second substrate layer comprises one or more of;

a polymer, plastic, glass, quartz, and silicon.

27. The method of claim 1, wherein the body comprises a first substrate material having the at least two intersecting microchannels etched, molded or stamped in the surface of the substrate, the body further comprising a planar cover layer, which cover layer overlays and encloses at least a portion of the at least two intersecting channels.

28. The method of claim 1, wherein the microfluidic device further comprises a sample channel which extends to at least the surface of the body structure, which sample channel is fluidly coupled to the at least two microchannels.

29. The method of claim 1, wherein the microfluidic device further comprises a capillary pipettor which is coupled to at least one of the at least two microchannels.

30. The method of claim 1, wherein the at least two microchannels are arranged as part of a serial input parallel reaction channel configuration.

31. The method of claim 30, wherein the microfluidic device further comprises between about 10 and about 500 reaction channels.

32. The method of claim 1, wherein the microfluidic device flusher comprises a particle retention region fluidly coupled to the at least two microchannels.

33. The method of claim 32, further comprising a bead in the particle retention region.

34. The method of claim 1, the material transport system comprising an electrokinetic controller.

35. The method of claim 1, the material transport system comprising a pump.