High surface area substrates for microarrays and methods to make same

• US 20030148401A1
• Filed: 11/08/2002
• Published: 08/07/2003
• Est. Priority Date: 11/09/2001
• Status: Active Grant

• Current Assignee: 3Dbiosurfaces Technologies, LLC (Stratec SE)
• Original Assignee: 3dbiosurfaces, Inc.
• Inventors: Galbraith, David W., Agrawal, Anoop, Cronin, John P., Hogan, Michael E., LeCompte, Robert S., Goodyear, A. G., Tonazzi, Juan C.
• Granted Patent: US 7,195,872 B2
• Time in Patent Office: Days
• US Class Current: 435/7.9
• 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 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00378 Piezoelectric or ink jet dispensers
• CPC Class Codes: B01J 2219/00387 Applications using probes
• CPC Class Codes: B01J 2219/00504 Pins
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00574 radioactive
• CPC Class Codes: B01J 2219/00576 fluorophore
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00617 by chemical means
• CPC Class Codes: B01J 2219/00621 by physical means, e.g. trenches, raised areas
• CPC Class Codes: B01J 2219/00628 Ionic
• CPC Class Codes: B01J 2219/00635 by reactive plasma treatment
• CPC Class Codes: B01J 2219/00637 by coating it with another layer
• CPC Class Codes: B01J 2219/00644 the porous medium being present in discrete locations, e.g. gel pads
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00677 : Ex-situ synthesis followed by deposition on the substrate
• CPC Class Codes: B01J 2219/00722 : Nucleotides
• CPC Class Codes: B01J 2219/00725 : Peptides
• CPC Class Codes: B01J 2219/00734 : Lipids
• CPC Class Codes: B01J 2219/00743 : Cells
• CPC Class Codes: B01L 2200/12 : Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0819 : Microarrays; Biochips
• CPC Class Codes: B01L 2300/16 : Surface properties and coatings
• CPC Class Codes: B01L 2300/166 : Suprahydrophobic; Ultraphobic; Lotus-effect
• CPC Class Codes: B01L 3/5085 : for multiple samples, e.g. microtitration plates
• CPC Class Codes: B01L 3/5088 : confining liquids at a location by surface tension, e.g. virtual wells on plates, wires B01L3/50857 takes precedence
• CPC Class Codes: B81B 1/006 : Microdevices formed as a single homogeneous piece, i.e. wherein the mechanical function is obtained by the use of the device, e.g. cutters
• CPC Class Codes: B81B 2201/0214 : Biosensors; Chemical sensors
• CPC Class Codes: B81C 1/00206 : Processes for functionalising a surface, e.g. provide the surface with specific mechanical, chemical or biological properties

Abstract

The present invention is directed to a substrate having a plurality of microfeatures that provide a high surface area and are open to provide ready access to fluids and components therein. Methods of making the high surface area substrates are described and include generating microfeatures and/or microstructures on the surface of the substrate.

Claims

1. A substrate having a high surface area for use in a microarray comprising a plurality of microfeatures on a surface of the substrate arranged in spatially discrete regions to produce a texture on the surface, wherein the textured surface provides an increase in surface area as compared to a non-textured surface.

2. The substrate of claim 1, wherein the substrate comprises a material selected from the group consisting of glass, a ceramic, a metal, a non-metal and a polymer.

3. The substrate of claim 2, wherein the microfeatures comprise a material that is different from the material of the substrate, said material of the microfeatures is selected from the group consisting of a glass, a ceramic, a metal, a non-metal, an inorganic oxide and a polymer.

4. The substrate of claim 1, wherein the plurality of microfeatures comprise a pit, a trench, a pillar, a cone, a wall, a micro-rod, a tube, a channel or a combination thereof.

5. The substrate of claim 1, wherein the plurality of microfeatures comprise communicating microfeatures.

6. The substrate of claim 1, wherein the plurality of microfeatures are distributed uniformly on the surface of the substrate.

7. The substrate of claim 1, wherein the plurality of microfeatures are distributed randomly on the surface of the substrate.

8. The substrate of claim 1, wherein each of the plurality of microfeatures has an aspect ratio less than about 10.

9. The substrate of claim 8, wherein each of the plurality of microfeatures has a height of about 0.1 to about 100 microns.

10. The substrate of claim 8, wherein each of the plurality of microfeatures has a cross-section of about 0.01 to about 500 sq. microns.

11. The substrate of claim 1, wherein each of the plurality of microfeatures has an aspect ratio of less than about 5.

12. The substrate of claim 1, wherein each of the plurality of microfeatures has an aspect ratio of less than about 1.

13. The substrate of claim 1, wherein the textured surface is characterized by having a peak-to-valley value of greater than about 10 nm.

14. The substrate of claim 1, wherein the textured surface is characterized by having a peak-to-valley value of greater than about 100 nm.

15. The substrate of claim 1, wherein the textured surface is characterized by having a peak-to-valley value of greater than about 1000 nm.

16. The substrate of claim 1, wherein the surface area is increased by at least about 20%.

17. The substrate of claim 1, wherein the surface area is increased by at least about 100%.

18. The substrate of claim 1, wherein the surface further comprises a plurality of microstructures.

19. The substrate of claim 18, wherein the plurality of microstructures comprise a pit, a trench, a pillar, a cone, a wall, a micro-rod, a tube, a channel or a combination thereof.

20. The substrate of claim 18, wherein the each of the plurality of microstructures are smaller in dimension than each of the plurality of microfeatures and occupy a spatially discrete region within the microfeature.

21. The substrate of claim 1, wherein the microfeatures are etched into the surface of the substrate.

22. The substrate of claim 21, wherein the etching comprises corrosion, ablation, abrasion, reaction, impact, drilling or dissolving.

23. The substrate of claim 1, wherein the microfeatures are formed on the surface of the substrate.

24. The substrate of claim 23, wherein the microfeatures comprise an inorganic oxide selected from the group consisting of tungsten oxide, silica, zirconia, alumina, titania, tantala, zinc oxide, nickel oxide, magnesium oxide, calcium oxide, boron oxide, potassium oxide, sodium oxide, chromium oxide, tin oxide, lithium oxide, lead oxide, and phosphorous oxide.

25. The substrate of claim 1, wherein at least one spatially discrete region comprises a well, said well comprising at least two microfeatures and a plurality of microstructures that are each integral to the bottom of the well, said microstructures are smaller in dimension than each of the microfeatures.

26. The substrate of claim 1, wherein each of the plurality of microfeatures are smaller in dimension than an array element of the microarray, said array element comprises a textured array element comprised of at least two microfeatures.

27. The substrate of claim 26, wherein the textured array element provides local replications of measurements as compared to a non-textured array element.

28. The substrate of claim 26, wherein each of the local measurements in said textured array element is a uniform intensity as compared to a non-textured array element.

29. The substrate of claim 26, wherein at least one of said microfeatures of said textured array element functions as a well-defined boundary of said textured array element.

30. The substrate of claim 1, further comprising an activating material coated on at least one of the plurality of microfeatures.

31. The substrate of claim 30, wherein the activating material functions to immobilize a biomolecule.

32. The substrate of claim 31, wherein the biomolecule comprises at least one of a nucleic acid, an oligonucleotide, a peptide, a polypeptide, a protein, an enzyme, a cell, an organelle, a lipid, a carbohydrate, a fat, a vitamin, a nutrient, or an antibody.

33. The substrate of claim 31, further characterized by providing an access of the biomolecule to the textured surface that is substantially similar to an access provided by a non-textured surface.

34. The substrate of claim 30, wherein the activating material has an iso-electric point (IEP) equal to or greater than about 4.

35. The substrate of claim 30, wherein the activating material comprises a silane, a metal oxide, streptavidin, salicylhydroxamic acid (SHA), a hydrazine, an aldehyde, a ketone, an ester, an amide, poly(ethyleneimine), polyamide, polyacrylamide, nitrocellulose, an organic molecule having at least one free amino group, or derivatives thereof.

36. The substrate of claim 35, wherein the silane comprises an amino group, a mercaptan, or an epoxy group.

37. The substrate of claim 35, wherein the metal oxide comprises tungsten oxide, silica, zirconia, alumina, titania, tantala, zinc oxide, nickel oxide, magnesium oxide, calcium oxide, boron oxide, potassium oxide, sodium oxide, chromium oxide, tin oxide, lithium oxide, lead oxide, or phosphorous oxide.

47. A microarray comprising a high surface area substrate of claim 1.

48. A microarray comprising a high surface area substrate of claim 18.

76. A method of preparing the high surface area of claim 1 comprising molding the substrate and each of the plurality of microfeatures in one step using a molding material.

77. The method of claim 76, wherein the molding material comprises polycarbonate, acrylic, polystyrene, nylon, polyolefin, or silicone.

78. The method of claim 76, wherein at least one of the spatially discrete regions is a well comprising a textured bottom surface comprised of said plurality of microfeatures.

79. The method of claim 78, wherein the textured bottom surface further comprises a plurality of microstructures, wherein each of said plurality of microstructures are smaller in dimension than each of the plurality of microfeatures.

80. The method of claim 76, wherein said molding comprises injection molding or compression molding.

81. The method of claim 76, further comprising applying an activating material to at least one of the plurality of microfeatures, wherein the activating material functions to immobilize a biomolecule.

38. A substrate having a high surface area for use in a microarray comprising a well, said well comprising a textured bottom surface comprised of a plurality of microfeatures that provide the high surface area.

39. The substrate of claim 38, the textured bottom surface further comprising a plurality of microstructures, wherein each of said microstructures are smaller in dimension than each of the plurality of microfeatures.

40. The substrate of claim 38, wherein each of the plurality of microfeatures are smaller in dimension than an array element of the microarray, said array element comprises a textured array element comprised of at least two microfeatures.

41. A microarray having improved analytical accuracy comprising a high surface area substrate, said substrate comprising a plurality of microfeatures on a surface of the substrate arranged in spatially discrete regions to produce a texture on the surface, wherein the textured surface provides an improvement in accuracy as compared to a non-textured surface.

42. The microarray of claim 41, wherein each of the plurality of microfeatures are smaller in dimension than an array element of the microarray, said array element comprises a textured array element comprised of at least two microfeatures.

43. The microarray of claim 42, wherein the textured array element provides local replications of measurements.

44. The microarray of claim 42, wherein at least one of said microfeatures of the textured array element functions as a well-defined boundary of said textured array element.

45. A microarray having improved uniformity of signal intensity comprising a high surface area substrate, said substrate comprising a plurality of microfeatures on a surface of the substrate arranged in spatially discrete regions to produce a texture on the surface, wherein the textured surface provides the improvement in the uniformity of the signal intensity as compared to a non-textured surface.

46. The microarray of claim 45, wherein each of the plurality of microfeatures are smaller in dimension than an array element of the microarray, said array element comprises a textured array element comprised of at least two microfeatures.

84. A method of detecting an analyte comprising

applying a sample to a microarray of claim 46;

binding the sample to at least one biomolecule, wherein the biomolecule is a probe; and

detecting the binding, wherein the binding indicates the presence of the analyte.

85. The method of claim 84, wherein the probe is labeled with a fluorescent molecule, a particle, a chemiluminescent fragment, or a radioactive molecule.

86. The method of claim 84, wherein the step of detecting comprises detecting

a fluorescent signal, light scattering, a radioactive signal, an optical signal, an electronic signal, or mass desorption.

87. The method of claim 84, wherein detecting comprises electronic discrimination.

88. The method of claim 87, wherein the electronic discrimination comprises determining a change in mass, capacitance, resistance, inductance or a combination thereof as compared to a control.

89. The method of claim 84, wherein the analyte is selected from the group consisting of a small organic molecule, a biomolecule, a macromolecule, a particle and a cell.

49. A method of preparing a substrate having a high surface area for use in a microarray comprising the steps of:

covering a surface of the substrate with a coating material; and

,

patterning the coating material, said patterning defines a plurality of microfeatures arranged in spatially discrete regions on the surface to form a textured surface, wherein said textured surface provides the high surface area as compared to a non-textured surface.

50. The method of claim 49, wherein the coating material comprises a photoresist polymer, a metal, a non-metal, or a ceramic.

51. The method of claim 50, wherein the coating material further comprises an adhesion promoter.

52. The method of claim 49, wherein the patterning comprises photolithography.

53. The method of claim 49, wherein the patterning comprises embossing the coating material.

54. The method of claim 53, wherein the coating material is deposited on the surface by a sol-gel process.

55. The method of claim 54, wherein the coating material comprises an inorganic oxide comprised of tungsten oxide, silica, zirconia, alumina, titania, tantala, zinc oxide, nickel oxide, magnesium oxide, calcium oxide, boron oxide, potassium oxide, sodium oxide, chromium oxide, tin oxide, lithium oxide, lead oxide, or phosphorous oxide.

56. The method of claim 53, wherein the embossing comprises consolidation, said consolidation involves applying heat, radiation or a combination thereof of the embossed surface.

57. The method of claim 49, further comprising forming a plurality of micro-rods on the surface.

58. The material of claim 57, wherein the coating material inhibits the growth of micro-rods.

59. The method of claim 57, wherein the micro-rod has an aspect ratio of less than about 10.

60. The method of claim 57, wherein the micro-rod has an aspect ratio of less than about 5.

61. The method of claim 57, wherein the micro-rod has an aspect ratio of less than about 1.

62. The method of claim 57, wherein the micro-rod is hollow.

63. The method of claim 57, further comprising applying an activating material to at least one of the plurality of microfeatures, wherein the activating material functions to immobilize a biomolecule.

64. The method of claim 57, further comprising the step of removing the coating material from the textured surface.

65. The method of claim 64, further comprising applying an activating material to at least one of the plurality of microfeatures, wherein the activating material functions to immobilize a biomolecule.

66. The method of claim 49, further comprising the step of etching the textured surface, wherein said etching decreases an aspect ratio of said plurality of microfeatures.

67. The method of claim 66, wherein the coating material is etch-resistant.

68. The method of claim 66, wherein etching comprises corrosion, ablation, abrasion, reaction, impact, drilling, or dissolving.

69. The method of claim 66, wherein etching comprises a wet-chemical process.

70. The method of claim 66, wherein etching comprises anisotropic etching.

71. The method of claim 66, further comprising applying an activating material to at least one of the plurality of microfeatures, wherein the activating material functions to immobilize a biomolecule.

72. The method of claim 66, further comprising the step of removing the coating material from the textured surface.

73. The method of claim 72, further comprising applying an activating material to at least one of the plurality of microfeatures, wherein the activating material functions to immobilize a biomolecule.

74. The method of claim 49, further comprising applying an activating material to at least one of the plurality of microfeatures, wherein the activating material functions to immobilize a biomolecule.

75. The method of claim 74, wherein the biomolecule comprises at least one of a nucleic acid, an oligonucleotide, a peptide, a polypeptide, a protein, an enzyme, a cell, an organelle, a lipid, a carbohydrate, a fat, a vitamin, a nutrient, or an antibody.

82. A method of preparing a substrate having a high surface area for a microarray comprising embossing a surface of the substrate to generate a plurality of microfeatures on the surface to form a textured surface that provides the high surface area.

83. The method of claim 82, wherein the substrate comprises a well plate or a micro-well plate and each of said plurality of microfeatures occupy a bottom surface of at least one well of said well plate or said micro-well plate.

90. A bioreactor having an increased surface area comprising a substrate comprising at least one reservoir, said reservoir comprising more than one communicating microfeature, wherein said microfeature increases the surface area by at least about 100% as compared to a flat surface.

91. The bioreactor of claim 90, wherein the reservoir further comprises a microstructure, said microstructure is smaller in dimension than the microfeature.

92. The bioreactor of claim 90, wherein the reservoir comprises a reaction chamber, said reaction comprises surface catalysis, mixing, filtration, or fractionation.

93. A substrate having a high surface area comprising

a solid substrate;

and

a layer of a coating on a surface of the substrate comprising an inorganic oxide and a plurality of microchannels, wherein said microchannels are formed from a removable fibrous template.

94. The substrate of claim 93, wherein the fibrous template comprises a plurality of particles having a fiber aspect ratio of at least about 3.

95. The substrate of claim 93, wherein the fibrous template comprises a peptide, latex, collagen, a glycol, an amine or an organic acid.

96. The substrate of claim 93, wherein the fibrous template further comprises a removable non-fibrous template that forms a void in the coating.

97. The substrate of claim 96, wherein the non-fibrous template comprises a peptide, latex, collagen, a glycol, an amine or an organic acid.

98. The substrate of claim 93, wherein the inorganic oxide comprises tungsten oxide, silica, zirconia, alumina, titania, tantala, zinc oxide, nickel oxide, magnesium oxide, calcium oxide, boron oxide, potassium oxide, sodium oxide, chromium oxide, tin oxide, lithium oxide, lead oxide, or phosphorous oxide.

99. The substrate of claim 93, wherein the coating is formed by a sol-gel process.

100. A substrate for use in a microarray comprising:

a coating which is doped with a dopant, wherein the coating is selected from the group consisting of silica, titania, tantala, zirconia, tantala, tungsten oxide and alumina, and wherein the dopant is selected from the group consisting of lithium oxide, sodium oxide , potassium oxide, zinc oxide, and organic salts.

101. The substrate of claim 100 wherein the coating is silica and the dopant is lithium oxide or zinc oxide.

102. The substrate in claim 100, where said coating is porous.

103. The substrate of claim 100 wherein the coating is formed by a wet chemical deposition process.

104. The substrate of claim 103, wherein said wet chemical deposition process comprises deposition of a liquid precursor comprising an organic moiety.

105. The substrate of claim 104 wherein the organic moiety is ablated.

106. A method of preparing a substrate for use in a microarray having a high surface area and having a plurality of surface microfeatures, comprising the step of ion-etching the surface of the substrate.

107. The method of claim 106 wherein the step of ion-etching process comprises reactive ion-etching using a reactive ion.

108. The method of claim 107 wherein the step of ion-etching uses a reactive ion formed from carbon tetraflouride gas.

109. The method of claim 107 wherein the reactive ion uses a reactive species generated by an inductively coupled plasma.

110. The method of claim 106 further comprising at least a second ion etching step utilizing a different reactive ion than that used initially in said step of ion etching.

111. The method of claim 110 wherein said different reactive ion is formed from oxygen or argon.

112. The method of claim 106 further comprising the step of etching and plasma treatment using an activating gas.

113. The method of claim 112 wherein said activating gas is ammonia.

114. The method of claim 110 further comprising the step of etching and plasma treatment using an activating gas.

115. A microarray formed from a substrate produced by the process of claim 106 wherein each array element occupies more than one microfeature.