Integrated solid-phase hydrophilic matrix circuits and micro-arrays

• US 20030127333A1
• Filed: 12/02/2002
• Published: 07/10/2003
• Est. Priority Date: 06/04/2001
• Status: Active Grant

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Epocal Incorporated (Siemens AG)
• Inventors: Bergevin, Benoit R., Wojtyk, James, Pierce, Raymond J., Lauks, Imants
• Granted Patent: US 7,201,833 B2
• Time in Patent Office: Days
• US Class Current: 204/600
• CPC Class Codes: B01J 2219/00286 Reactor vessels with top and bottom openings
• CPC Class Codes: B01J 2219/00358 electrode driven
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B01J 2219/00783 Laminate assemblies, i.e. the reactor comprising a stack of plates
• CPC Class Codes: B01J 2219/00831 Glass
• CPC Class Codes: B01J 2219/00853 Employing electrode arrangements
• CPC Class Codes: B01J 2219/0086 Dimensions of the flow channels
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0825 Test strips
• CPC Class Codes: B01L 2300/10 Means to control humidity and/or other gases
• 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 3/5023 with a sample being transported to, and subsequently stored in an absorbent for analysis
• 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 3/50273 : 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: G01N 2035/00158 : Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 33/6845 : Methods of identifying protein-protein interactions in protein mixtures
• CPC Class Codes: Y02E 60/50 : Fuel cells

Abstract

The invention is directed to analytical devices and micro-arrays with integral fluidic inputs and outputs. The devices are constructed from planar solid-phase hydrophilic matrix circuits containing dry chemical reagents overlaying integral electro-kinetic pumping electrodes. The hydrophilic matrix circuits are enclosed within a gas permeable electrical insulator. The devices are for use in micro-scale bio-analysis, mixture separation and reaction.

Claims

1. An enclosed hydrophilic matrix device for transport of an aqueous solute, comprising

an electrically insulated substrate;

a hydrophilic matrix path on the substrate for electro-kinetic transport of the solute, the matrix path having a pair of spaced apart contacting locations for respective electric contact with a source of power for producing an electric potential along the hydrophilic matrix path;

an electrode supported on the substrate and having a contact end for connection to the power source and a matrix end for electric contact with the hydrophilic matrix at one of the contacting locations;

the matrix being initially dry and including a humectant for increasing a water absorption rate of the matrix;

an insulator enclosing the hydrophilic matrix for sealing the matrix between the insulator and the substrate, the insulator being water vapor permeable; and

an orifice in the insulator above the matrix for the passage of an aqueous solute through the insulator.

2. The device of claim 1, including a pair of electrodes supported on the substrate, each electrode having a contact end for connection to the power source and a matrix end for electric connection to the matrix at one of the contacting locations.

3. The device of claim 1, wherein the hydrophilic matrix is initially in a dry and inactive state in which it is substantially non-conductive, and is transferred into a humidified state by transport of water vapor through the insulator upon exposure of the enclosing insulator to an aqueous environment.

4. The device of claim 3, wherein the insulator is gas permeable to permit incorporation of water into the matrix by capillary action through the orifice.

5. The device of claim 3, wherein the insulator includes a wet-up opening for the passage of water and is gas permeable to permit venting of gas within the enclosed matrix during incorporation of water into the matrix by capillary action through the wet-up opening.

6. The device of claim 3, wherein water is incorporated into the matrix by transport through the water vapor permeable material of the insulator.

7. The device of claim 1, wherein the humectant is a low molecular weight neutral molecule which when dissolved in water forms an aqueous solution with a water vapor pressure significantly less than pure water at a concentration where the solution'"'"'s viscosity is not significantly higher than pure water.

8. The device of claim 6, wherein the humectant is selected from the group of urea, alanine, orthinine, praline, lysine, glycine, polyols and sugars:

sucrose, glucose, xylitol, sorbitol, mannitol, lactose, maltose, lactulose, glycerol, propylene glycol, citric acid, tartaric acid, malic acid and combinations thereof.

9. The device of claim 2, wherein electric contact between the matrix and the electrodes at the contact locations is achieved by direct physical contact between the electrode and matrix materials at the contacting locations.

10. The device of claim 2, wherein the electrodes and matrix path are spaced apart at the contact locations and electric contact is achieved by an intermediate conductive substance.

11. The device of claim 2, wherein the electrodes and matrix path are spaced apart at the contact locations and electric contact is achieved by a hydrophilic intermediate conductive substance which is initially in a dry and non conductive condition when the matrix is in the dry condition and is rendered conductive upon wet up of the matrix.

12. The device of claim 2, wherein the electrodes and matrix path are spaced apart at the contact locations and electric contact is achieved by a hydrophilic substance which is included in the matrix at the contacting locations and is initially in a dry and non conductive condition when the matrix is in the dry condition, is rendered conductive upon wet up of the matrix and electrically bridges the space between the electrodes and the matrix at the contacting locations after wet-up.

13. The device of claim 1, wherein the hydrophilic matrix path has a fixed charge for electro-osmotic transport of the aqueous solute therethrough.

14. The device of claim 1, wherein the hydrophilic matrix contains a reagent to be electro-kinetically pumped through the orifice.

15. The device of claim 13, wherein the reagent is in a dry state when the matrix is in the dry state, the reagent in the dry state being substantially positionally and chemically stable.

16. The device of claim 1, wherein the hydrophilic matrix contains electrolyte salt.

17. The device of claim 16, wherein the maximum electrolyte salt concentration in the matrix is 10 mM.

18. The device of claim 1, wherein the humectant is a neutral molecule.

19. The device of claim 17, wherein the neutral humectant is loaded to give a wet-up concentration of greater than 1 molar in the humidified state of the matrix.

20. The device of claim 1, wherein the hydrophilic matrix further contains a redox additive.

21. The device of claim 19, wherein the redox additive is neutral.

22. The device of claim 1, wherein the hydrophilic matrix is micro-porous.

23. The device of claim 21, wherein micro-pores of the matrix have a diameter between 50 nanometers and 5 micrometers.

24. The device of claim 1, wherein the hydrophilic matrix has a maximum thickness of 50 micrometers.

25. The device of claim 1, wherein the water vapor permeable insulator is less than 25 micrometers in thickness.

26. The device of claim 1, wherein one of the pair of electrodes is constructed as a cathode electrode for supporting an oxygen reduction reaction, and the enclosing insulator is gas permeable for permitting lateral diffusion of oxygen through the insulator.

27. The device of claim 1, wherein one of the pair of electrodes is constructed as an anode electrode for supporting a water oxidation reaction, and the enclosing insulator is gas permeable for permitting oxygen removal from the electrode region by lateral permeation through the gas permeable insulator.

28. The device of claim 1, wherein the hydrophilic matrix material is selected to be dry-etchable.

29. The device of claim 27, wherein the hydrophilic matrix further contains dry-etchable additives.

30. The device of claim 1, wherein the hydrophilic matrix path further includes a reservoir for containing a reagent to be transported along the matrix path by electrokinetic transport.

31. The device of claim 29, wherein the hydrophilic matrix reservoir region is circular and contains reagents locally deposited from a micro-nozzle dispenser, ink jet dispenser or a pin-transfer dispenser.

32. The device of claim 29, wherein the transport path includes an air gap located between the matrix reservoir and the orifice in the insulator.

33. The device of claim 29, the matrix further including a second hydrophilic matrix reservoir interposed between the other electrode and the orifice.

36. A planar array of hydrophilic matrix fluidic i/o devices, comprising an array of micro-locations each including a hydrophilic matrix fluidic i/o device as defined in claim 29.

40. A bioassay device, comprising in combination an enclosed hydrophilic matrix fluidic i/o device according to claim 29 and a reactor device as defined in claim 35, the orifice in the insulator of the hydrophilic matrix fluidic i/o device overlapping the orifice in the insulator of the reactor device for reagent exchange and the hydrophilic matrix fluidic i/o device being constructed for electro-kinetically transporting the reagent from the reservoir to the reactor device through the orifice.

41. A bioassay device, comprising

a first planar array in accordance with claim 40 and having the micro-reactor devices arranged at preselected step-and-repeat dimensions;

a second planar array of micro-locations each including an immobilized reactant and being arranged at the same step-and-repeat dimensions as the micro-reactor devices in the first array;

alignment means for aligning the co-planar first and second arrays in a spaced apart parallel orientation so that the micro-reactors on the respective arrays are aligned pairwise opposite to one another;

means for introducing fluid between the co-planar first and second arrays; and

means for sealing each pair of micro-reactor and opposite micro-location for forming an array of isolated, fluid-filled wells, each well containing a micro-reactor of the first array, a spaced apart parallel micro-location of the second array and intermediate fluid.

42. A bioassay device, comprising

a first planar array in accordance with claim 37 and having the micro-reactor devices arranged at preselected step-and-repeat dimensions;

a second planar array in accordance with claim 29 and having the hydrophilic matrix fluidic i/o devices arranged at the same step-and-repeat dimensions as the micro-reactors in the first array;

alignment means for aligning the co-planar first and second arrays in a spaced apart parallel orientation so that each micro-reactor on the first planar array is opposite one hydrophilic matrix fluidic i/o device of the second array;

means for introducing fluid between the co-planar first and second arrays; and

means for sealing each pairing of micro-reactor and opposite hydrophilic matrix fluidic i/o device for forming an array of isolated, fluid-filled wells, each well containing a micro-reactor of the first array, a spaced apart parallel hydrophilic matrix fluidic i/o device of the second array and intermediate fluid.

43. The bioassay device of claim 41 or 42, further comprising means for monitoring a reaction in each of the isolated wells.

54. The device of claim 2, wherein the substrate has a pair of opposite surfaces, the matrix path is supported on one of the substrate surfaces and at least one of the pair of electrodes is supported on the other substrate surface, the substrate being shaped and constructed for providing electrical contact of the matrix with the electrode on the opposite substrate surface.

55. The device of claim 54, wherein the substrate includes a passage for physical and electrical contact of the matrix at one of the contacting locations with the electrode on the opposite substrate surface.

56. The device of claim 54, wherein the pair of electrodes are supported on one of the substrate surfaces and the matrix is supported on the opposite of the substrate surfaces and the substrate at each of the contacting locations has a throughgoing passage, the matrix material extending through the passage and into contact with the respective electrode.

34. The device of claim 58, wherein the second reservoir contains a reagent to be electro-kinetically pumped through the orifice.

35. A micro reactor device with integrated fluidic i/o, comprising

an insulated substrate;

a pair of electrodes supported on the substrate, each electrode having a contact end for connection to an external circuit for supplying power and a matrix end for electric contact with a hydrophilic matrix;

a hydrophilic matrix path on the substrate for electro-kinetic transport of the solute, the matrix path including a reservoir for containing a reagent, a transport path for electro-kinetic transport of the reagent, a discrete micro-reactor for carrying out a chemical reaction and a pair of spaced apart contacting locations for electric contact with the respective matrix ends of the electrodes, the matrix being initially dry and including a humectant for increasing a water absorption rate of the matrix;

an insulator enclosing the hydrophilic matrix for sealing the matrix between the insulator and the substrate, the insulator being water vapor permeable; and

an orifice in the insulator above the matrix for the passage of an aqueous solute through the insulator.

37. A planar micro-reactor array, comprising an array of micro-locations, each including a micro-reactor device as defined in claim 35.

38. The array of claim 37, wherein each reactor device is constructed to carry out a nucleic acid hybridization reaction.

39. The array of claim 37, wherein each reactor device is constructed to carry out a protein-protein interaction.

44. A micro reactor device as defined in claim 35, including a plurality of reservoirs each containing a different reagent for transport to the micro-reactor.

45. A micro reactor device as defined in claim 35, including a plurality of micro-reactors for receiving the reagent from the reservoir.

46. A micro reactor device as defined in claim 35, wherein the orifice is located between the reservoir and the micro-reactor.

47. A micro reactor device as defined in claim 46, wherein the matrix path includes a first portion extending between the contacting locations and a second portion in extension of the first portion, the device further comprising a gap in the matrix path for preventing osmotic transport thereacross of the reagent in the reservoir, the gap being located in the first portion and between the reservoir and the micro reactor.

48. A micro reactor device as defined in claim 35, wherein the matrix path includes a first portion extending between the contacting locations and a second portion in extension of the first portion, the reservoir being positioned in the second portion and the orifice being located in the second portion between the reservoir and the first portion.

49. A micro reactor device as defined in claim 35, wherein the matrix path includes in series a first portion, a second portion and a third portion, the second portion extending between the contacting locations, the reservoir being located in the first portion and the orifice and the micro-reactor being located in the third portion.

50. A micro reactor device as defined in claim 49, further comprising a second reservoir located in the second portion.

51. A micro reactor device as defined in claim 50, further comprising a gap in the matrix path in one of the second and third portions for preventing osmotic transport of the solute thereacross, the gap being located between the second reservoir and the orifice.

52. A micro reactor device as defined in claim 51, wherein the gap is located in the second portion between the reservoir and the third portion.

53. A bioassay device, comprising a first micro reactor device as defined in claim 35 and a second micro reactor device as defined in claim 35 arranged in series to the first micro reactor device such that the matrix path of the second micro reactor device is separated from the matrix path of the first micro reactor device by an intermediate air gap which can be bridged through electrokinetic pumping along the matrix path of the first micro reactor device.