Substrate comprising a nano-patterned surface and method for preparing the same

Abstract

Disclosed are substrates and the preparation of the substrates by using a room temperature UV nanoembossing method, wherein the substrate comprises: a functionalizable layer, a polymer layer comprising a plurality of micrometer-scale patterns or nanometer-scale patterns or a combination thereof, and a backing layer. The substrates can be prepared by a roll-to-roll continuous process. The substrates can be used as flow cells, nanofluidic devices or microfluidic devices for biomolecular analysis.

Description

[0001] Related Applications

[0002] This application is a divisional application of Chinese Patent Application No. 201480064276.4, and claims the benefit of priority of U.S. Provisional Application No. 61 / 918,582 filed on December 19, 2013, which is incorporated herein by reference in its entirety. Field of the Invention

[0003] In general, the present application relates to the field of nanopatterning methods and substrates comprising nanopatterned surfaces. More specifically, the present application relates to substrates comprising at least two layers and in some embodiments at least three layers, the substrate comprising a polymer layer comprising nanoscale patterns and micrometer-scale patterns. Methods for preparing these substrates by using a resist-free, room temperature UV curing and embossing method are also disclosed. Background of the Invention

[0004] Nanoprinting techniques are used to enable economical and efficient production of nanostructures. Standard nanoembossing lithography relies on direct mechanical deformation of a resist material by a stamp with nanostructures, followed by an etching process to transfer the nanostructures from the stamp to the substrate. SUMMARY OF THE INVENTION

[0005] The present application utilizes UV sensitive polymers to form a polymer layer containing multiple micrometer-scale patterns or nanometer-scale patterns. The UV release process prevents the polymer from sticking to the template and prevents any deformation of the pattern during the separation process, thus giving a stable pattern transfer even at the nanometer level. The method of the present application is complementary to the hot embossing method and does not require heating of the master template, and thus avoids problems associated with thermal stress in the material and thermal gradients across the template.

[0006] The method of the present application can be applied to technologies where it is necessary to carry out low-cost micro-patterning and nano-patterning. Applications include nanomanufacturing, medical diagnosis and global health, flexible display technology, nanofluids, and emerging markets including construction and automotive industries, where low-cost methods for producing nano-patterned, super-hydrophobic surfaces (dewetting windows, windshields, etc.) in emerging markets gain traction. The method of the present application can also be applied to produce low-cost patterned consumables, for example, patterned plastic flow cells for sequencing devices and low-cost microfluidic devices.

[0007] With the spiraling cost of healthcare and the lack of medical infrastructure in developing countries, there is a growing demand for point-of-care (POC) diagnostics and low-cost lab-on-a-chip (LOC) technologies. These technologies can gain expanded applicability if they are made cheaper, stable, portable, disposable, and most importantly, provide a fast time to produce results. To make such progress possible, low-cost patterning and microfluidic technologies are needed. Paper-based microfluidics have emerged as a potential candidate, but they are difficult to manufacture using conventional lithography-based methods, difficult to perform multiplexed detection of multiple analytes, and easily rely on capillary action to carry away solutions, so pre-enrichment of target analytes is required in many embodiments. The method of the present application provides a highly cost-effective option for manufacturing a total microfluidic-based analytical platform for global health applications.

[0008] Some embodiments disclosed herein include a substrate comprising: a functionalizable layer comprising one or more functional groups; a polymer layer comprising a plurality of microscale patterns or nanoscale patterns, or a combination thereof; and a backing layer.

[0009] In some embodiments, the functionalizable layer is arranged between the backing layer and the polymer layer. In some embodiments, the polymer layer is arranged between the backing layer and the functionalizable layer. In some embodiments, the substrate can also include a sealing layer to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer. In some such embodiments, the sealing layer is optically transparent.

[0010] In some embodiments, at least one of the microscale pattern or the nanoscale pattern can allow entry of a sample fluid. In some embodiments, the substrate can also include a fluid reservoir, for example, a sample reservoir.

[0011] In any disclosed embodiment, the functionalizable layer may comprise a reactive silane layer, a functionalizable hydrogel, or a functionalizable polymer. In some such embodiments, the functionalizable layer may comprise one or more functional groups. In some such embodiments, the functional group may be selected from: optionally substituted olefins, azides, optionally substituted amines, carboxylic acids, optionally substituted hydrazones, halogens, hydroxyls, optionally substituted tetrazoles, optionally substituted tetrazines, thiols, and combinations thereof.

[0012] In any disclosed embodiment, the functional group of the functionalizable layer can be connected to a biomolecule. In some embodiments, the biomolecule is selected from amino acids, nucleosides, nucleotides, peptides, oligonucleotides, polynucleotides, nucleic acids, proteins, or combinations thereof. In some such embodiments, the biomolecule is a polynucleotide or a nucleic acid.

[0013] In any disclosed embodiment, the polymer layer comprises at least one photocurable polymer. In some embodiments, the photocurable polymer comprises urethane, acrylate, siloxane, epoxy, polyacrylic acid, polyacrylate, epoxysiloxane, epoxy resin, polydimethylsiloxane (PDMS), silsesquioxane, acyloxysilane, maleic acid polyester, vinyl ether, monomers containing vinyl or ethynyl groups, or copolymers and combinations thereof.

[0014] Some embodiments disclosed herein include methods for preparing a substrate, the method comprising: providing a substrate including a functionalizable layer comprising one or more functional groups, wherein the functionalizable layer is disposed between a backing layer and a photocurable polymer layer; contacting the surface of the photocurable polymer layer with a template having a plurality of micrometer-scale patterns or nanometer-scale patterns including micrometer-scale or nanometer-scale holes, pillars, or a combination thereof; applying pressure to the template or substrate to transfer the micrometer-scale pattern or nanometer-scale pattern to at least the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature; irradiating the photocurable polymer with UV light to cure the photocurable polymer; and separating the template from the substrate; wherein at least a portion of the polymer layer is penetrated to expose the functionalizable layer below. In some embodiments, at least a portion of the polymer layer and the functionalizable layer is penetrated to expose the backing layer below.

[0015] Some embodiments disclosed herein include methods for preparing a substrate, comprising: providing a template comprising a plurality of micrometer-scale patterns or nanometer-scale patterns including micrometer-scale or nanometer-scale holes, channels, or combinations thereof on the surface of the template; depositing a functional layer comprising one or more functional groups on the surface of the template so that at least a portion of the micrometer-scale or nanometer-scale holes, channels, or combinations thereof comprises a functionalized layer; removing excess functionalizable layer from the surface of the template so that the functionalizable layer is present only on a portion of the surface of the template; providing a substrate comprising a backing layer, wherein the backing layer has a photocurable polymer layer disposed on the backing layer; contacting the surface of the photocurable polymer layer with the surface of the template having a plurality of micrometer-scale patterns or nanometer-scale patterns and a functionalizable layer thereon; applying pressure to the template or substrate to transfer the micrometer-scale patterns or nanometer-scale patterns to at least the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature; irradiating the photocurable polymer with UV light to cure the photocurable polymer; and separating the template from the substrate; wherein at least a portion of the functionalizable layer is transferred to the polymer layer. In some embodiments, at least a portion of the polymer layer is penetrated to expose the backing layer. In some embodiments, the substrate further comprises a backing layer having a photocurable polymer layer disposed on the backing layer, and the substrate is a roll of flexible dicing tape.

[0016] In any disclosed method embodiment, the method may also include applying a sealing layer to the substrate after removing the template to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer. In some embodiments, the sealing layer also comprises a second substrate prepared by the method of any disclosed embodiment, wherein the functionalizable layer of the substrate and the photocurable polymer layer are arranged between the backing layer of the substrate. In some embodiments, the sealing layer is optically transparent. In some method embodiments, the method may be a roll-to-roll method.

[0017] As non-limiting examples, the present application provides the following embodiments:

[0018] 1. A substrate comprising:

[0019] a functionalizable layer comprising one or more functional groups;

[0020] a polymer layer comprising a plurality of micro-scale patterns or nano-scale patterns or a combination thereof; and

[0021] Backing layer.

[0022] 2. The substrate of embodiment 1, wherein the microscale pattern or nanoscale pattern of the polymer layer comprises channels, grooves, holes, pillars, or a combination thereof.

[0023] 3. The substrate of embodiment 1 or 2, wherein the functionalizable layer is disposed between the backing layer and the polymer layer.

[0024] 4. The substrate of embodiment 3, wherein at least a portion of the microscale pattern or nanoscale pattern of the polymer layer penetrates through the polymer layer to expose the functionalizable layer underneath.

[0025] 5. A substrate as described in embodiment 4, wherein the functionalizable layer comprises a plurality of microscale patterns or nanoscale patterns or a combination thereof, and at least a portion of the microscale patterns or nanoscale patterns of the polymer layer and the functionalizable layer penetrate through the polymer layer and the functionalizable layer to expose the backing layer underneath.

[0026] 6. The substrate of embodiment 1 or 2, wherein the polymer layer is disposed between the backing layer and the functionalizable layer.

[0027] 7. The substrate of embodiment 6, wherein the functionalizable layer comprises a plurality of microscale patterns or nanoscale patterns or a combination thereof, wherein at least a portion of the microscale patterns or nanoscale patterns of the functional layer penetrates through the functionalizable layer to expose the polymer layer underneath.

[0028] 8. The substrate of embodiment 7, wherein at least a portion of the microscale pattern or nanoscale pattern of the functionalizable layer and the polymer layer penetrates through the functionalizable layer and the polymer layer to expose the backing layer underneath.

[0029] 9. The substrate of embodiment 6, wherein at least a portion of the micro-scale pattern or nano-scale pattern of the polymer layer penetrates to expose the backing layer underneath.

[0030] 10. The substrate of any one of embodiments 1 to 9, wherein the substrate is in the shape of a cylinder, and wherein the backing layer is closer to the outer surface of the cylinder than the functionalizable layer or polymer layer of the substrate.

[0031] 11. The substrate of any one of embodiments 1 to 9, wherein the substrate is in the shape of a cylinder, and wherein the backing layer is closer to the inner surface of the cylinder than the functionalizable layer or polymer layer of the substrate.

[0032] 12. The substrate of any one of embodiments 1 to 11, wherein the substrate further comprises a sealing layer to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer.

[0033] 13. The substrate of embodiment 12, wherein the sealing layer is optically transparent.

[0034] 14. The substrate of any one of embodiments 1 to 13, wherein at least one of the microscale pattern or the nanoscale pattern is capable of allowing entry of a sample fluid.

[0035] 15. The substrate of any one of embodiments 1 to 14, further comprising a fluid reservoir.

[0036] 16. The substrate of any one of embodiments 1 to 15, wherein at least a portion of the microscale pattern or nanoscale pattern is a columnar body.

[0037] 17. The substrate of embodiment 16, wherein the average diameter of the columns is less than about 500 nm.

[0038] 18. The substrate of embodiment 17, wherein the average diameter of the columns is about 330 nm.

[0039] 19. The substrate of any one of embodiments 16 to 18, wherein the average height of the columns is less than about 500 nm.

[0040] 20. The substrate of embodiment 19, wherein the average height of the columns is about 300 nm.

[0041] 21. The substrate of any one of embodiments 1 to 20, wherein the backing layer is made of a material selected from the group consisting of silica, plastic, quartz, metal, metal oxide, paper, and combinations thereof.

[0042] 22. The substrate of embodiment 21, wherein the backing layer is made of a flexible plastic material.

[0043] 23. The substrate of embodiment 21 or 22, wherein the backing layer is made from a roll of flexible film.

[0044] 24. The substrate of any one of embodiments 1 to 23, wherein the functionalizable layer comprises a reactive silane layer, a functionalizable hydrogel, or a functionalizable polymer.

[0045] 25. The substrate of any one of embodiments 1 to 24, wherein the functionalizable layer comprises one or more functional groups.

[0046] 26. The substrate of embodiment 25, wherein the functional group is selected from the group consisting of optionally substituted olefins, azides, optionally substituted amines, carboxylic acids, optionally substituted hydrazones, hydroxyls, optionally substituted tetrazoles, optionally substituted tetrazines, thiols, and combinations thereof.

[0047] 27. The substrate of any one of embodiments 1 to 26, wherein the functionalizable layer comprises a polymer or hydrogel comprising formula (Ia) or formula (Ib):

[0048]

[0049] R 1 is H or optionally substituted alkyl;

[0050] Functional group R A is selected from the group consisting of: azide, optionally substituted amine, optionally substituted olefin, optionally substituted hydrazone, carboxylic acid, halogen, hydroxyl, optionally substituted tetrazole, optionally substituted tetrazine, and thiol;

[0051] R 5 is selected from H or optionally substituted alkyl;

[0052] Each -(CH2)-p can be optionally substituted;

[0053] P is an integer from 1 to 50;

[0054] n is an integer from 1 to 50,000; and

[0055] m is an integer from 1 to 100,000.

[0056] 28. The substrate of any one of embodiments 25 to 27, wherein the functional group comprises an azide.

[0057] 29. The substrate of any one of embodiments 25 to 28, wherein the functional groups of the functionalizable layer are linked to biomolecules.

[0058] 30. The substrate of embodiment 29, wherein the biomolecule is selected from amino acids, nucleosides, nucleotides, peptides, oligonucleotides, polynucleotides, nucleic acids, proteins, or combinations thereof.

[0059] 31. The substrate of embodiment 30, wherein the biomolecule is a polynucleotide or a nucleic acid.

[0060] 32. The substrate of any one of embodiments 1 to 31, wherein the polymer layer comprises at least one photocurable polymer.

[0061] 33. A substrate as described in embodiment 32, wherein the photocurable polymer comprises: ethyl formate, acrylate, siloxane, epoxy, polyacrylic acid, polyacrylate, epoxysiloxane, epoxy resin, polydimethylsiloxane (PDMS), silsesquioxane, acyloxysilane, maleic acid polyester, vinyl ether, monomers containing vinyl or ethynyl groups, or copolymers and combinations of the above.

[0062] 34. The substrate of any one of embodiments 1 to 33, wherein the backing layer has a thickness of no more than about 1 mm.

[0063] 35. The substrate of any one of embodiments 1 to 34, wherein the backing layer has a thickness of no more than about 100 μm.

[0064] 36. The substrate of any one of embodiments 1 to 35, wherein the thickness of the functionalizable layer does not exceed about 1 mm.

[0065] 37. The substrate of any one of embodiments 1 to 36, wherein the functionalizable layer has a thickness of no more than about 100 μm.

[0066] 38. The substrate of any one of embodiments 1 to 37, wherein the polymer layer has a thickness of no more than about 1 mm.

[0067] 39. The substrate of any one of embodiments 1 to 38, wherein the polymer layer has a thickness of no more than about 100 μm.

[0068] 40. The substrate of any one of embodiments 1 to 39, wherein at least one of the backing layer, polymer layer, or functionalizable layer is further doped with a dopant material.

[0069] 41. A substrate as described in embodiment 40, wherein the dopant material is selected from: nanoparticles, quantum dots, binding receptors, ligands, nucleic acids, reactive parts in the functionalizable layer, filter materials, lighter absorbing materials, luminescent materials, light scattering materials, conductive materials or thermally conductive materials.

[0070] 42. The substrate of embodiment 40 or 41, wherein the dopant material is present primarily or only on the surface of the backing layer, polymer layer, or functionalizable layer.

[0071] 43. The substrate of embodiment 40 or 41, wherein the dopant material is present within the volume of the backing layer, polymer layer, or functionalizable layer.

[0072] 44. A substrate as described in embodiment 40 or 41, wherein the dopant material is present both within the volume of the backing layer, polymer layer or functionalizable layer and on the surface of the backing layer, polymer layer or functionalizable layer.

[0073] 45. The substrate of any one of embodiments 1 to 44, wherein the plurality of microscale patterns or nanoscale patterns or a combination thereof are made by mechanical embossing at room temperature.

[0074] 46. ​​A method for preparing a substrate, comprising:

[0075] providing a substrate comprising a functionalizable layer comprising one or more functional groups, wherein the functionalizable layer is disposed between a backing layer and a photocurable polymer layer;

[0076] contacting the surface of the photocurable polymer layer with a template having a plurality of micro-scale patterns or nano-scale patterns including micro-scale or nano-scale holes, pillars, or a combination thereof;

[0077] applying pressure to the template or substrate to transfer the microscale pattern or nanoscale pattern to at least the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature;

[0078] irradiating the photocurable polymer with UV light to cure the photocurable polymer; and

[0079] separating the template from the substrate;

[0080] At least a portion of the polymer layer is penetrated to expose the functionalizable layer underneath.

[0081] 47. The method of embodiment 46, wherein at least a portion of the polymer layer and the functionalizable layer are penetrated to expose the backing layer underneath.

[0082] 48. A method for preparing a substrate, comprising:

[0083] Providing a template comprising a plurality of micro-scale patterns or nano-scale patterns including micro-scale or nano-scale holes, channels, or a combination thereof on a surface of the template;

[0084] depositing a functional layer comprising one or more functional groups on the surface of the template so that at least a portion of the micro- or nano-scale pores, channels, or a combination thereof comprises the functional layer;

[0085] removing excess functionalizable layer from the surface of the template so that the functionalizable layer is present only on a portion of the template surface;

[0086] providing a substrate including a backing layer having a photocurable polymer layer disposed on the backing layer;

[0087] contacting the surface of the photocurable polymer layer with the surface of the template having a plurality of micro-scale patterns or nano-scale patterns and a functionalizable layer thereon;

[0088] applying pressure to the template or substrate to transfer the microscale pattern or nanoscale pattern to at least the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature;

[0089] irradiating the photocurable polymer with UV light to cure the photocurable polymer; and

[0090] separating the template from the substrate;

[0091] wherein at least a portion of the functionalizable layer is transferred to the polymer layer.

[0092] 49. The method of embodiment 48, wherein at least a portion of the polymer layer is penetrated to expose the backing layer.

[0093] 50. The method of embodiment 48 or 49, wherein the substrate is a roll of flexible dicing tape, the substrate comprising the backing layer having a photocurable polymer layer disposed on the backing layer.

[0094] 51. The method of any one of embodiments 46 to 50, further comprising applying a sealing layer to the substrate after removing the template so as to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer.

[0095] 52. A method as described in embodiment 51, wherein the sealing layer includes a second substrate prepared by a method as described in any one of embodiments 46 to 49, wherein the functionalizable layer and the photocurable polymer layer of the substrate are arranged between the backing layer of the substrate.

[0096] 53. The method of any one of embodiments 51 and 52, wherein the sealing layer is optically transparent.

[0097] 54. The method of any one of embodiments 46 to 53, further comprising forming the substrate into a cylinder.

[0098] 55. The method of embodiment 54, wherein the backing layer is closer to the outer surface of the cylinder than the functionalizable layer or polymer layer of the substrate.

[0099] 56. The method of embodiment 54, wherein the backing layer is closer to the inner surface of the cylinder than the functionalizable layer or polymer layer of the substrate.

[0100] 57. The method of any one of embodiments 46 to 56, wherein at least a portion of the microscale pattern or nanoscale pattern is a column.

[0101] 58. A method as described in embodiment 57, wherein the average diameter of the pillars is less than about 500 nm.

[0102] 59. A method as described in embodiment 58, wherein the average diameter of the pillars is about 330 nm.

[0103] 60. The method of any one of embodiments 57 to 59, wherein the average height of the pillars is less than about 500 nm.

[0104] 61. A method as described in embodiment 60, wherein the average height of the columns is about 300 nm.

[0105] 62. The method of any one of embodiments 46 to 61, wherein the backing layer is made of a material selected from the group consisting of silica, plastic, quartz, metal, metal oxide, paper, and combinations thereof.

[0106] 63. The method according to embodiment 62, wherein the backing layer is made of a flexible plastic material.

[0107] 64. The method of embodiment 62 or 63, wherein the backing layer is made from a roll of flexible film.

[0108] 65. The method of any one of embodiments 46 to 64, wherein the functionalizable layer comprises a reactive silane layer, a functionalizable hydrogel, or a functionalizable polymer.

[0109] 66. The method of any one of embodiments 46 to 65, wherein the functionalizable layer comprises one or more functional groups.

[0110] 67. A method as described in embodiment 66, wherein the functional group is selected from: optionally substituted olefins, azides, optionally substituted amines, carboxylic acids, optionally substituted hydrazones, hydroxyls, optionally substituted tetrazoles, optionally substituted tetrazines, thiols, and combinations thereof.

[0111] 68. The method of any one of embodiments 46 to 67, wherein the functionalizable layer comprises a polymer or hydrogel comprising Formula (Ia) or Formula (Ib):

[0112]

[0113] R 1 is H or optionally substituted alkyl;

[0114] Functional group R A is selected from the group consisting of: azide, optionally substituted amine, optionally substituted olefin, optionally substituted hydrazone, carboxylic acid, halogen, hydroxyl, optionally substituted tetrazole, optionally substituted tetrazine, and thiol;

[0115] R 5 is selected from H or optionally substituted alkyl;

[0116] Each -(CH2)-p can be optionally substituted;

[0117] p is an integer from 1 to 50;

[0118] n is an integer from 1 to 50,000; and

[0119] m is an integer from 1 to 100,000.

[0120] 69. The method of any one of embodiments 66 to 68, wherein the functional group comprises an azide.

[0121] 70. The method of any one of embodiments 66 to 69, wherein the functional group of the functionalizable layer is linked to a biomolecule.

[0122] 71. The method of embodiment 70, wherein the biomolecule is selected from an amino acid, a nucleoside, a nucleotide, a peptide, an oligonucleotide, a polynucleotide, a nucleic acid, a protein, or a combination thereof.

[0123] 72. A method as described in embodiment 71, wherein the biomolecule is a polynucleotide or a nucleic acid.

[0124] 73. The method of any one of embodiments 46 to 72, wherein the polymer layer comprises at least one photocurable polymer.

[0125] 74. A method as described in embodiment 73, wherein the photocurable polymer comprises urethane, acrylate, siloxane, epoxy, polyacrylic acid, polyacrylate, epoxysiloxane, epoxy resin, polydimethylsiloxane (PDMS), silsesquioxane, acyloxysilane, maleic acid polyester, vinyl ether, monomers containing vinyl or ethynyl groups, or copolymers and combinations of the above.

[0126] 75. The method of any one of embodiments 46 to 74, wherein the thickness of the backing layer does not exceed about 1 mm.

[0127] 76. The method of any one of embodiments 46 to 75, wherein the backing layer has a thickness of no more than about 100 μm.

[0128] 77. The method of any one of embodiments 46 to 76, wherein the thickness of the functionalizable layer does not exceed about 1 mm.

[0129] 78. The method of any one of embodiments 46 to 77, wherein the thickness of the functionalizable layer does not exceed about 100 μm.

[0130] 79. The method of any one of embodiments 46 to 78, wherein the polymer layer has a thickness of no more than about 1 mm.

[0131] 80. The method of any one of embodiments 46 to 79, wherein the polymer layer has a thickness of no more than about 100 μm.

[0132] 81. The method of any one of embodiments 46 to 80, wherein at least one of the backing layer, polymer layer, or functionalizable layer is also doped with a dopant material.

[0133] 82. A method as described in embodiment 81, wherein the dopant material is selected from: nanoparticles, quantum dots, binding receptors, ligands, nucleic acids, reactive parts in the functionalizable layer, filter materials, lighter absorbing materials, luminescent materials, light scattering materials, conductive materials or thermally conductive materials.

[0134] 83. The method of embodiment 81 or 82, wherein the dopant material is present primarily or only on the surface of the backing layer, polymer layer, or functionalizable layer.

[0135] 84. The method of embodiment 81 or 82, wherein the dopant material is present within the volume of the backing layer, polymer layer, or functionalizable layer.

[0136] 85. A method as described in embodiment 81 or 82, wherein the dopant material is present both within the volume of the backing layer, polymer layer or functionalizable layer and on the surface of the backing layer, polymer layer or functionalizable layer.

[0137] 86. A method as described in any of embodiments 81 to 85, wherein at least one of the backing layer, the photocurable polymer layer, or the functionalizable layer is doped with a dopant material prior to performing the method as described in embodiment 46 or 48.

[0138] 87. A method as described in any of embodiments 81 to 86, wherein at least one of the photocurable polymer layer or the functionalizable layer is doped with a dopant material before contacting the template according to the method of embodiment 46 or 48.

[0139] 88. The method of any one of embodiments 81 to 87, wherein at least a portion of the microscale pattern or nanoscale pattern of the template is transferred to the dopant material in the polymer layer.

[0140] 89. The method of any one of embodiments 81 to 87, wherein at least a portion of the microscale pattern or nanoscale pattern of the template is transferred to the dopant material in the functionalizable layer.

[0141] 90. The method of any one of embodiments 46 to 89, wherein the method is a roll-to-roll method. BRIEF DESCRIPTION OF THE DRAWINGS

[0142] Figures 1A-1C are cross-sectional views of several embodiments of substrates in which a functionalizable layer is disposed between a polymeric layer and a backing layer.

[0143] Figure 2A-2C are cross-sectional views of several embodiments of substrates in which a functionalizable layer is disposed between a polymeric layer and a backing layer.

[0144] Figure 2D is a top view of a substrate with a functionalizable layer disposed between a polymer layer and a backing layer.

[0145] Figure 2E is a cross-sectional view of a substrate in which a functionalizable layer and a polymer layer are disposed between a backing layer and a sealing layer.

[0146] Figures 3A-3C are cross-sectional views of several embodiments of substrates in which a polymeric layer is disposed between a functionalizable layer and a backing layer.

[0147] Figure 4A and 4Bare cross-sectional views of several embodiments of substrates in which a functionalizable layer is disposed between a polymeric layer and a backing layer.

[0148] Figure 4C is a cross-sectional view of a substrate in which a functionalizable layer and a polymer layer are disposed between a backing layer and a sealing layer.

[0149] Figure 5A and 5B are cross-sectional views of several embodiments of substrates in the shape of a cylinder.

[0150] Fig. 6A and 6B are cross-sectional views of several embodiments of substrates in the shape of a cylinder.

[0151] Fig. 7A and 7B are cross-sectional views of several embodiments of two substrates stacked on top of one another with the distal surfaces of the two substrates (ie, surfaces distal from their respective substrate layers) in contact or in close proximity.

[0152] Figures 8A-8D A cross-sectional view of an embodiment of a method of preparing a substrate in which a functionalizable layer is disposed between a polymer layer and a backing layer using a template comprising a plurality of micro-scale patterns or nano-scale patterns is described.

[0153] Figures 9A-9E A cross-sectional view of an embodiment of a method of preparing a substrate in which a polymer layer is disposed between a functionalizable layer and a backing layer using a template comprising a plurality of micro-scale patterns or nano-scale patterns is described.

[0154] Fig. 10A A top view of an 8-inch master template wafer containing 12 Hi-seq-sized patterned flow cells.

[0155] Fig. 10B Is from Fig. 10A Dark field microscope image of a top view of a cut-away flow cell 7.

[0156] Figures 10C-10F The top view SEM images of the nanoscale patterns formed on the polymer sheet at 1240X, 20kX, 40kX and 80kX magnifications, respectively.

[0157] Fig.11A and 11B The top view SEM images of the nanoscale patterns at 1307x and 5000x magnifications, respectively.

[0158] Fig. 12A is a top-view optical image of the serpentine microfluidic channels and intersections.

[0159] Fig. 12B is a top view of an optical image of a microfluidic filter.

[0160] Fig. 12C is a top view of an optical image of fluid flow through a serpentine microfluidic channel as a function of time.

[0161] Fig.12D is a top view of the optical image of the fluid flowing into the T-junction.

[0162] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0163] The following detailed description relates to some specific embodiments of the present application. In this description, with reference to the accompanying drawings, for clarity, the same parts or steps throughout the text may be designated with similar numbers. "One embodiment", "embodiment", or "in some embodiments" mentioned in this specification means that the specific features, structures or characteristics described in conjunction with the embodiment may be included in at least one embodiment of the present invention. The appearance of the phrase "one embodiment", "embodiment", or "in some embodiments" in different places of this specification does not necessarily refer to all the same embodiments, nor is it a separate or alternative embodiment mutually exclusive with other embodiments. In addition, various features that can be expressed by some embodiments but not by other embodiments are described. Similarly, various requirements that may be some embodiment requirements but not other embodiment requirements are described.

[0164] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0165] definition

[0166] Unless otherwise defined, all technical terms and scientific terms used herein have the same meaning as those generally understood by those skilled in the art. Unless otherwise stated, all patents, applications, published applications and other publications cited herein are incorporated by reference as a whole. As used in this specification and the appended claims, unless otherwise clearly indicated in the context, the singular forms "a / one (a)", "an" and "the" include plural indicators. Unless otherwise stated, the use of "or" means "and / or". In addition, the use of the term "including" and other forms, such as "including / include", "includes" and "included" is not restrictive. As used in this specification, whether in transition words or in the body of the claims, the term "comprise(s)" and "comprising" should be understood to have an open meaning. That is, the term should be understood to be synonymous with the phrase "at least having" or "at least including". When used in the context of a method, the term "comprising" means that the method includes at least the recited steps, but may include additional steps. When used in the context of a compound, composition, or device, the term "comprising" means that the compound, composition, or device includes at least the recited features or components, but may also include additional features or components.

[0167] As used herein, common organic abbreviations are defined as follows:

[0168] PAZAM Poly(N-(5-(2-azidoacetamidopentyl)acrylamide-co-acrylamide) with any ratio of acrylamide to Azapa (N-(5-(2-azidoacetamidopentyl)acrylamide)

[0169] SEM Scanning Electron Microscope

[0170] ℃ Temperature in degrees Celsius

[0171] μm micrometer

[0172] As used herein, the term "array" refers to different probe molecule populations connected to one or more substrates so that different probe molecules can be spatially distinguished from each other. The array can include different probe molecules, each of which is located at different addressable positions on the substrate. Alternatively or in addition, the array can include a separate substrate, each of which has different probe molecules, wherein the different probe molecules can be identified according to the position of the substrate on the surface connected to the substrate or according to the position of the substrate in the liquid. The exemplary array of the single substrate positioned on the surface includes, but is not limited to, those including beads (beads) in the hole, as described in the PCT disclosure of No. 6,355,431B1, No. US 2002 / 0102578 and No. WO 00 / 63437. For example, in No. 6,524,793 U.S. Patent, it is described that can be used for the present application to, for example, utilize a microfluidic device such as a fluorescence activated cell sorter (FACS) to distinguish the exemplary form of beads in the liquid array. Additional examples of arrays that can be used in the present application include, but are not limited to, U.S. Pat. Nos. 5,429,807; 5,436,327; 5,561,071; 5,583,211; 5,658,734; 5,837,858; 5,874,219; 5,919,523; 6,136,269; 6,287,768; 6,287,776; 6,288,220; 6,297,006; 6,291,193; 6,346,413; 6,416,949; 6,482,591; 6,514,751 and 6,610,482; and WO 93 / 17126; WO 95 / 11995; WO 95 / 35505; EP 742 287; and those described in EP 799 897.

[0173] As used herein, the term "covalently attached" or "covalent bonding" refers to the formation of a chemical bond characterized by the sharing of electron pairs between atoms. For example, when a "covalently attached polymer coating" is used to refer to a substrate surface, it refers to a polymer coating that forms a chemical bond with the functionalized surface of the substrate, as opposed to being attached to the surface by other means such as adhesion or electrostatic interactions. It should be understood that a polymer covalently attached to a surface may also be bonded by other means besides covalent attachment.

[0174] As used herein, "cutting tape" refers to a flexible substrate comprising a backing layer and an adhesive layer. The adhesive layer can be treated, for example, by thermal treatment or photophysical treatment (e.g., UV light) to suppress or destroy bonding properties. Cutting tape can be made of PVC, polyolefin, or polyethylene backing materials containing adhesive to fix the mold in place. Cutting tape can have various thicknesses, for example, from 75 μm to 150 μm, and have various bond strengths. UV tape is a cutting tape in which bonding force is destroyed by exposure to UV light after cutting.

[0175] As used herein, a "functionalizable" layer refers to a layer or coating comprising a reactive portion that can be connected to one or more biomolecules by chemical reaction or intermolecular interaction. Such connection can be via a covalent bond or by other binding or interaction forces. In some embodiments, intermolecular interaction can be a specific binding between a ligand and a receptor, and paired ligands and receptors include, but are not limited to, streptavidin and biotin, nucleic acids and their complements, antibodies and ligands, and other known in the art. For example, a functionalizable layer can be a hydrogel comprising one or more functional groups that can react with or bind to a target biomolecule. A non-limiting specific example is a PAZAM comprising one or more azide functional groups that can react with an oligonucleotide comprising an alkynyl group. In some cases, a functionalizable layer becomes a functionalized layer by leaving reactive sites in reaction with a target biomolecule.

[0176] As used herein, the term "photocurable polymer" refers to a polymer that is capable of undergoing polymerization when exposed to actinic radiation (eg, UV radiation).

[0177] As used herein, the term "roll-to-roll process" refers to the processing of an elongated substrate as it is transferred from one reel to another. An exemplary roll-to-roll process is to continuously pattern a surface with micrometer-scale patterns or nanometer-scale patterns as the surface moves through a patterning device while being unwound from one reel and wound onto another reel.

[0178] As used herein, "alkyl" refers to a fully saturated (i.e., containing no double or triple bonds) straight or branched hydrocarbon chain. An alkyl group may have 1 to 20 carbon atoms (whenever it appears in this article, a numerical range such as "1 to 20" refers to each integer in the given range; for example, "1 to 20 carbon atoms" means that the alkyl group may consist of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms, however, this definition also encompasses the occurrence of the term "alkyl" in which no numerical range is specified). An alkyl group may be a medium-sized alkyl group having 1 to 9 carbon atoms. An alkyl group may also be a lower alkyl group having 1 to 4 carbon atoms. An alkyl group may be designated as "C1-4 alkyl" or similar designation. By way of example only, "C 1-4 "Alkyl" means that there are one to four carbon atoms in the alkyl chain, i.e., the alkyl chain is selected from methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl. Typical alkyl groups include, but are by no means limited to, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, and the like.

[0179] As used herein, "alkene" or "alkenyl" refers to a straight or branched hydrocarbon chain containing one or more double bonds. The alkenyl group may have from 2 to 20 carbon atoms, however, this definition also encompasses occurrences of the term "alkenyl" where no numerical range is specified. Alkenyl groups may also be medium-sized alkenyl groups having from 2 to 9 carbon atoms. Alkenyl groups may also be lower alkenyl groups having from 2 to 4 carbon atoms. Alkenyl groups may be designated as "C 2-4 By way of example only, "C 2-4 The term "alkenyl" means that there are two to four carbon atoms in the alkenyl chain, i.e., the alkenyl chain is selected from the group consisting of ethenyl, propen-1-yl, propen-2-yl, propen-3-yl, buten-1-yl, buten-2-yl, buten-3-yl, buten-4-yl, 1-methyl-propen-1-yl, 2-methyl-propen-1-yl, 1-ethyl-ethen-1-yl, 2-methyl-propen-3-yl, buta-1,3-dienyl, buta-1,2,-dienyl and buta-1,2-dien-4-yl. Typical alkenyl groups include, but are by no means limited to, ethenyl, propenyl, butenyl, pentenyl and hexenyl, etc.

[0180] As used herein, "alkynyl" refers to a straight or branched hydrocarbon chain containing one or more triple bonds. Alkynyl groups can have from 2 to 20 carbon atoms, however, this definition also encompasses occurrences of the term "alkynyl" where no numerical range is specified. Alkynyl groups can also be medium-sized alkynyl groups having from 2 to 9 carbon atoms. Alkynyl groups can also be lower alkynyl groups having from 2 to 4 carbon atoms. Alkynyl groups can be designated as "C 2-4 "alkynyl" or similar designation. By way of example only, "C 2-4 "Alkynyl" means that there are two to four carbon atoms in the alkynyl chain, that is, the alkynyl chain is selected from: ethynyl, propyn-1-yl, propyn-2-yl, butyn-1-yl, butyn-3-yl, butyn-4-yl and 2-butynyl. Typical alkynyl groups include, but are by no means limited to: ethynyl, propynyl, butynyl, pentynyl and hexynyl, etc.

[0181] As defined herein, "amino" refers to "-NR A R B " group, where R A and R B Each independently selected from: hydrogen, C 1-6 Alkyl, C2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclic group, C 6-10 Aryl, 5-10 membered heteroaryl, and 5-10 membered heterocyclyl. Non-limiting examples include free amino (ie, -NH2).

[0182] As used herein, "aryl" refers to an aromatic ring or aromatic ring system (i.e., two or more fused rings that share two adjacent carbon atoms) that contains only carbon in the ring backbone. When aryl is a ring system, each ring in the system is aromatic. Aryl can have 6 to 18 carbon atoms, however, this definition also encompasses occurrences of the term "aryl" where no numerical range is specified. In some embodiments, aryl has 6 to 10 carbon atoms. Aryl can be designated as "C 6-10 Aryl", "C6 or C 10 Examples of aryl groups include, but are not limited to, phenyl, naphthyl, azulenyl, and anthracenyl.

[0183] As used herein, the term "carboxylic acid" as used herein refers to -C(O)OH.

[0184] As used herein, "carbocyclyl" means a non-aromatic ring or non-aromatic ring system that contains only carbon atoms in the ring system backbone. When a carbocyclyl is a ring system, two or more rings may be joined together in a fused, bridged or spiro manner. A carbocyclyl may have any degree of saturation, provided that at least one ring in the ring system is not aromatic. Thus, carbocyclyl includes cycloalkyl, cycloalkenyl and cycloalkynyl. A carbocyclyl may have 3 to 20 carbon atoms, however this definition also encompasses the occurrence of the term "carbocyclyl" in which no numerical range is specified. A carbocyclyl may also be a medium-sized carbocyclyl having 3 to 10 carbon atoms. A carbocyclyl may also be a carbocyclyl having 3 to 6 carbon atoms. A carbocyclyl may be designated as "C 3-6 Examples of carbocyclyl rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, 2,3-dihydro-indene, bicyclo[2.2.2]octyl, adamantyl, and spiro[4.4]nonyl.

[0185] As used herein, the term "hydrazone" or "hydrazone group" as used herein refers to Group, where R a and R b are each independently selected from the group consisting of: hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-7 Carbocyclic group, C 6-10Aryl, 5-10 membered heteroaryl and 5-10 membered heterocyclyl. Non-limiting examples include free amino (ie, -NH2).

[0186] As used herein, the term "halogen" or "halo" means any of the radiostable atoms of column 7 of the periodic table, for example, fluorine, chlorine, bromine or iodine, preferably fluorine and chlorine.

[0187] As used herein, "heteroaryl" refers to an aromatic ring or aromatic ring system (i.e., two or more fused rings sharing two adjacent atoms) containing one or more heteroatoms in the ring skeleton, wherein the heteroatoms are elements other than carbon, including but not limited to nitrogen, oxygen and sulfur. When heteroaryl is a ring system, each ring in the system is aromatic. Heteroaryl can have 5-18 ring members (i.e., the number of atoms (including carbon atoms and heteroatoms) constituting the ring skeleton), but this definition also encompasses the occurrence of the term "heteroaryl" in which no numerical range is specified. In some embodiments, heteroaryl has 5 to 10 ring members or 5 to 7 ring members. Heteroaryl can be designated as "5-7 yuan heteroaryl", "5-10 yuan heteroaryl" or similar designations. Examples of heteroaryl rings include, but are not limited to, furanyl, thienyl, phthalazinyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, triazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, quinolyl, isoquinolyl, benzimidazolyl, benzoxazolyl, benzothiazolyl, indolyl, isoindolyl, and benzothiophenyl.

[0188] As used herein, "heterocyclic radical" means a non-aromatic ring or non-aromatic ring system containing at least one heteroatom in the ring skeleton. The heterocyclic radical can be connected together in a fused, bridged or spiro manner. The heterocyclic radical can have any degree of saturation, provided that at least one ring in the ring system is not aromatic. The heteroatom can be present in a non-aromatic ring or an aromatic ring in the ring system. The heterocyclic radical can have 3 to 20 ring members (i.e., the number of atoms (including carbon atoms and heteroatoms) constituting the ring skeleton), but the present definition also encompasses the occurrence of the term "heterocyclic radical" in which no numerical range is specified. The heterocyclic radical can also be a medium-sized heterocyclic radical with 3 to 10 ring members. The heterocyclic radical can also be a heterocyclic radical with 3 to 6 ring members. The heterocyclic radical can be designated as "3-6-membered heterocyclic radical" or similar designations. In preferred six-membered monocyclic heterocyclyls, the heteroatoms are selected from one up to three of O, N or S, and in preferred five-membered monocyclic heterocyclyls, the heteroatoms are selected from one or two heteroatoms of O, N or S. Examples of heterocyclyl rings include, but are not limited to, azepinyl, acridinyl, carbazolyl, cinnolinyl, dioxolanyl, imidazolinyl, imidazolidinyl, morpholinyl, oxiranyl, oxepanyl, thiepanyl, piperidinyl, piperazinyl, dioxopiperazinyl, pyrrolidinyl, pyrrolidinone, pyrrolidinedionyl, 4-piperidone, pyrazolinyl, pyrazolidinyl, 1,3-dioxinyl, 1,3-dioxinyl, 1,4-dioxinyl, 1,4-dioxinyl, 1,3-oxathianyl, 1,4-oxathianyl, 1,4-oxathianyl, 2H-1,2-oxazinyl, trialkyl( 1,3-dioxolyl, 1,3-dioxolanyl, 1,3-dithiolanyl, 1,3-dithiolanyl, isoxazolinyl, isoxazolidinyl, oxazolinyl, oxazolidinyl, oxazolidinone, thiazolinyl, thiazolidinyl, 1,3-oxathiolanyl, indolinyl, isoindolinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydro-1,4-thiazinyl, thiomorpholinyl, dihydrobenzofuranyl, benzimidazolidinyl, and tetrahydroquinoline.

[0189] As used herein, the term "tetrazine" or "tetrazinyl" refers to a six-membered heteroaryl group containing four nitrogen atoms. The tetrazine may be optionally substituted.

[0190] As used herein, the term "tetrazolyl" or "tetrazolyl" refers to a five-membered heterocyclic group containing four nitrogen atoms. Tetrazolyl may be optionally substituted.

[0191] As used herein, "nucleotides" include nitrogenous heterocyclic bases, sugars, and one or more phosphate groups. They are monomeric units of nucleic acid sequences. In RNA, the sugar is ribose, and in DNA the sugar is deoxyribose, i.e., a sugar lacking the hydroxyl group present in the 2' position of the ribose. Nitrogenous heterocyclic bases can be purine bases or pyrimidine bases. Purine bases include adenine (A) and guanine (G), and modified derivatives or analogs thereof. Pyrimidine bases include cytosine (C), thymine (T) and uracil (U), and modified derivatives or analogs thereof. The C-1 atom of deoxyribose is bonded to the N-1 of pyrimidine or the N-9 of purine.

[0192] As used herein, "nucleoside" is similar in structure to nucleotides, but lacks any phosphate moiety at the 5' position. The term "nucleoside" is used herein in the common sense understood by those skilled in the art. Examples include, but are not limited to, ribonucleosides comprising a ribose moiety, and deoxyribonucleosides comprising a deoxyribose moiety. A modified pentose moiety is a pentose moiety in which an oxygen atom is replaced by a carbon and / or a carbon is replaced by a sulfur atom or an oxygen atom. A "nucleoside" is a monomer that may have a substituted base and / or sugar moiety. In addition, nucleosides may be incorporated into larger DNA and / or RNA polymers and oligomers.

[0193] As used herein, the term "polynucleotide" generally refers to nucleic acids, including DNA (e.g., genomic DNA cDNA), RNA (e.g., mRNA), synthetic oligonucleotides and synthetic nucleic acid analogs. Polynucleotides can include natural bases or non-natural bases or combinations thereof, and natural or non-natural backbone connections (e.g., phosphorothioate, PNA or 2'-O-methyl-RNA) or combinations thereof.

[0194] As used herein, the substituted group is derived from an unsubstituted parent group in which one or more hydrogen atoms are replaced by another atom or group. Unless otherwise indicated, when a group is considered to be "substituted", it means that the group is substituted by one or more substituents independently selected from the following: C1-C6 alkyl, C1-C6 alkenyl, C1-C6 alkynyl, C1-C6 heteroalkyl, C3-C7 carbocyclyl (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy), C3-C7-carbocyclyl-C1-C6-alkyl (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy) , 5-membered to 10-membered heterocyclic group (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy), 5-membered to 10-membered heterocyclic group-C1-C6-alkyl (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy), aryl (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy), aryl (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy), aryl (C1-C6) alkyl (optionally substituted by halogen, C1-C6 alkyl, C1-C6 alkoxy alkyl, C1-C6 haloalkyl and C1-C6 haloalkoxy optionally substituted), 5- to 10-membered heteroaryl (optionally substituted with halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy optionally substituted), 5- to 10-membered heteroaryl (optionally substituted with halogen, C1-C6 alkyl, C1-C6 alkoxy, C1-C6 haloalkyl and C1-C6 haloalkoxy optionally substituted), halogen, cyano, hydroxy, C1-C6 alkoxy, C1-C6 alkoxy(C1-C6)alkyl (i.e., ether), aryloxy, sulfhydryl (sulfh The invention also includes but is not limited to alkyl, alkyl, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene, alkylene,

[0195] Some embodiments disclosed herein include a substrate comprising: a functionalizable layer comprising one or more functional groups; a polymer layer comprising a plurality of microscale patterns or nanoscale patterns or a combination thereof; and a backing layer.

[0196] In some embodiments, the functionalizable layer is disposed between the backing layer and the polymer layer. In some such embodiments, at least a portion of the micrometer-scale pattern or nanometer-scale pattern of the polymer layer penetrates the polymer layer to expose the functionalizable layer below. In some such embodiments, the functionalizable layer comprises a plurality of micrometer-scale patterns or nanometer-scale patterns, or a combination thereof, and at least a portion of the micrometer-scale pattern or nanometer-scale pattern of the polymer layer and the functionalizable layer penetrates the polymer layer and the functionalizable layer to expose the backing layer below.

[0197] Figures 1A-1C 1 shows a cross-sectional view of a substrate 100 according to some embodiments of the present invention, the substrate 100 comprising: a functionalizable layer 102 comprising one or more functional groups; a polymer layer 103 comprising a plurality of micro-scale patterns or nano-scale patterns, or a combination thereof; and a backing layer 101, wherein the functionalizable layer 102 is disposed between the backing layer 101 and the polymer layer 103. Figure 1A As shown, substrate 100 comprises at least three layers: backing layer 101, functionalizable layer 102, and polymer layer 103. Polymer layer 103 comprises a plurality of nanoscale patterns 104 and 105 of different shapes and depths. In some other embodiments, the nanoscale patterns of the polymer layer may be the same.

[0198] In some embodiments, the polymer layer has a combination of both microscale patterns and nanoscale patterns. Figure 1B As shown, the polymer layer 103 further comprises a plurality of micrometer-scale patterns 106 and 107, wherein the nanometer-scale patterns 104 and 105 are formed within the micrometer-scale patterns. Figure 1C As shown, the nanoscale structure can be both nanopores 104 and nanocolumns 108. Figures 1A-1C In the embodiment shown in FIG. 1 , the underlying functionalizable layer 102 is not exposed.

[0199] In some embodiments, at least a portion of the micro-scale pattern or nano-scale pattern of the polymer layer penetrates through the polymer layer to expose the functionalizable layer underneath. Figure 2A A cross-sectional view of a substrate 200 according to an embodiment of the present invention is shown, the substrate 200 having Figures 1A-1C The substrate 200 has the same layer structure as described in the above, wherein the polymer layer 203 comprises nanoscale patterns 204 and 205. The nanoscale pattern 204 penetrates the polymer layer 203 to expose the functionalizable layer 202 below.

[0200] In some embodiments, the functionalizable layer comprises a plurality of microscale patterns or nanoscale patterns, or a combination thereof, and at least a portion of the polymer layer and the microscale patterns or nanoscale patterns of the functionalizable layer penetrate through the polymer layer and the functionalizable layer to expose the underlying backing layer. Figure 2B A substrate 200 according to another embodiment of the present invention is shown, wherein the functionalizable layer 202 comprises a plurality of nanoscale patterns 204 and 206 , and the nanoscale patterns 206 penetrate through the polymer layer 203 and the functionalizable layer 202 to expose the backing layer 201 therebelow.

[0201] In some embodiments, such as Figure 2C As shown, each nanoscale pattern 204 of the polymer layer 203 penetrates the polymer layer to expose the functionalizable layer 202 underneath.

[0202] Figure 2D Shows Figure 2B , wherein the nanoscale pattern 204 exposes the underlying functionalizable layer 202, and the nanoscale pattern 206 penetrates the polymer layer and the functionalizable layer to expose the underlying backing layer 201.

[0203] In some embodiments, the substrate may further comprise a sealing layer to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer. Figure 2E Shown is a cross-sectional view of a substrate according to an embodiment of the present invention with sealing layer 207 substantially sealing polymer layer 203 and functionalizable layer 202 between sealing layer 207 and backing layer 201 .

[0204] In some embodiments, the sealing layer can be optically transparent. In some embodiments, there can be some space between the sealing layer and the polymer layer to allow sample fluid to flow through.

[0205] In some embodiments, the polymer layer is disposed between the backing layer and the functionalizable layer.

[0206] Figures 3A-3C A cross-sectional view of a substrate 300 of some embodiments of the present invention is shown, the substrate 300 comprising: a functionalizable layer 302 comprising one or more functional groups; a polymer layer 303 comprising a plurality of micrometer-scale patterns or nanometer-scale patterns, or a combination thereof; and a backing layer 301, wherein the polymer layer 303 is disposed between the functionalizable layer 302 and the backing layer 301.

[0207] like Figure 3AAs shown, substrate 300 comprises at least three layers: backing layer 301, functionalizable layer 302 and polymer layer 303, wherein polymer layer 303 is disposed between functionalizable layer 302 and backing layer 301. Polymer layer 303 comprises a plurality of nanoscale patterns 304 (nano-pillars) and 305 (nano-holes).

[0208] In some other embodiments, the nanoscale pattern of the polymer layer can be uniform in size and shape.

[0209] In some embodiments, the polymer layer has a combination of both microscale patterns and nanoscale patterns. Figure 3B As shown, the polymer layer 303 further includes a plurality of micrometer-scale patterns 306 and 307. Figure 3A and 3B In the embodiment, the underlying polymer layer 303 is not exposed.

[0210] Figure 3C Shows Figure 3A 300 , wherein excess functionalizable layer 302 has been removed, except for that present in nanoscale patterns 304 and 305 .

[0211] In some embodiments, the functionalizable layer comprises a plurality of micro-scale patterns or nano-scale patterns, or a combination thereof, wherein at least a portion of the micro-scale patterns or nano-scale patterns of the functionalized layer penetrates through the functionalizable layer to expose the underlying polymer layer. Figure 4A A cross-sectional view of a substrate 400 having a functionalizable layer 402 including a nanoscale pattern 404 and wherein the nanoscale pattern 404 penetrates the functionalizable layer 402 to expose the polymer layer 403 underneath is shown in accordance with an embodiment of the present invention.

[0212] In some such embodiments, at least a portion of the microscale pattern or nanoscale pattern of the functionalizable layer and the polymer layer penetrates through the functionalizable layer and the polymer layer to expose the underlying backing layer. Figure 4B A substrate 400 of an embodiment of the present invention is shown, wherein a functionalizable layer 402 and a polymer layer 403 include a plurality of nanoscale patterns 406 that penetrate through the polymer layer 403 and the functionalizable layer 402 to expose the underlying backing layer 401, while nanoscale patterns 405 do not penetrate through the polymer layer 403. In some embodiments, the polymer layer itself may include a nanoscale pattern that is penetrated to expose the underlying backing layer 401.

[0213] In some embodiments, the substrate may further comprise a sealing layer to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer. Figure 4CAs shown, sealing layer 407 substantially seals polymer layer 403 and functionalizable layer 402 between sealing layer 407 and backing layer 401 .

[0214] In some embodiments, the sealing layer can be optically transparent. In some embodiments, there can be some space between the sealing layer and the functionalizable layer to allow sample fluid to flow through.

[0215] In some embodiments, the substrate is cylindrical. In some such embodiments, the backing layer is closer to the outer surface of the cylinder than the functionalized layer or polymer layer of the substrate. In some such embodiments, the backing layer is closer to the inner surface of the cylinder than the functionalized layer or polymer layer of the substrate. In some embodiments, the sample fluid can flow through the inner surface of the cylindrical substrate along the longitudinal axis of the cylindrical substrate.

[0216] Figure 5A , 5B 6A and 6B are radial cross-sectional views of cylindrical substrates 500 and 600 according to some embodiments of the present invention. Figure 5A As shown, the backing layer 501 is closer to the outer surface of the cylinder than the functionalizable layer 502 or the polymer layer 503 of the substrate 500. In addition, the functionalizable layer 502 includes a plurality of nanoscale patterns 504 and 505. The nanoscale pattern 504 penetrates the polymer layer 503 to expose the functionalizable layer 502 below. The nanoscale pattern 505 penetrates the polymer layer 503 and the functionalizable layer 502 to expose the backing layer 501 below. Figure 5B In the embodiment of the present invention, the backing layer 501 is closer to the inner surface of the cylinder than the functionalizable layer 502 or the polymer layer 503 of the substrate 500 .

[0217] Similarly, Fig. 6A The substrate 600 of the embodiment of the present application is shown, and the backing layer 601 of the substrate 600 is closer to the outer surface of the cylinder than the functionalizable layer 602 or the polymer layer 603. In addition, the functionalizable layer 602 includes a plurality of nanoscale patterns 604 and 605. The nanoscale pattern 604 penetrates the functionalizable layer 602 to expose the polymer layer 603 below, and the nanoscale pattern 605 penetrates the polymer layer 603 and the functionalizable layer 602 to expose the backing layer 601 below. Figure 6B In the embodiment of the present application, the backing layer 601 of the substrate 600 is closer to the inner surface of the cylinder than the functionalizable layer 602 or the polymer layer 603 of the substrate 600 .

[0218] Fig. 7A and 7B700 is a cross-sectional view of some embodiments of the present application, the substrate 700 is formed of two substrates, wherein the polymer layer 703 and the functionalizable layer 702 are substantially sealed between two backing layers 701. Fig. 7A In the embodiment, substrate 700 is formed by stacking substrate 700a on the distal surface of substrate 700b so that the distal surfaces of two polymer layers 703 are in contact with each other (the distal surface is referred to as the backing layer). Both substrates 700a and 700b include nanoscale patterns 704 and 705 in polymer layer 703, wherein nanoscale pattern 705 penetrates polymer layer 703 to expose the functionalizable layer 702 below.

[0219] exist Figure 7B In the embodiment, a substrate 700 is formed by stacking one substrate 700a on the distal surface of another substrate 700a so that the distal surfaces of two functionalizable layers 702 are in close proximity to each other (e.g., in many embodiments, the distal surfaces are in direct contact with each other). The nanoscale pattern 706 penetrates the polymer layer 703 and the functionalizable layer 702 to expose the backing layer 701 below. The spacing between the two functionalizable layers 702 can allow sample fluid to flow through. One or more edges of the substrate 700 can be substantially sealed to prevent fluid leakage.

[0220] In any of the disclosed embodiments, the substrate can further comprise a sealing layer to substantially seal the polymer layer and the functionalizable layer between the backing layer and the sealing layer. In some such embodiments, the sealing layer is optically transparent.

[0221] In any of the disclosed embodiments, at least one of the micro-scale pattern or the nano-scale pattern is capable of allowing entry of a sample fluid.

[0222] In any of the disclosed embodiments, the substrate can also include a fluid reservoir, such as a sample reservoir, a reagent reservoir, or a waste reservoir.

[0223] In any disclosed embodiment, the micron-scale pattern or nanometer-scale pattern of the polymer layer includes a channel, a groove, a hole, a column or a combination thereof. In some embodiments, at least a portion of the micron-scale pattern or the nanometer-scale pattern is a column. In some such embodiments, the average diameter of the column is less than about 500nm. In some such embodiments, the average diameter of the column is about 330nm or less than about 330nm, including, for example, less than about 300nm, 200nm, 100nm or 50nm. In some such embodiments, the average height of the column is less than about 500nm. In some other embodiments, the average height of the column is about 300nm or less than about 300nm, including, for example, less than about 300nm, 200nm, 100nm or 50nm. Alternatively or additionally, for exemplary upper limits, the average diameter and / or average height of the pillars may be at most 1 mm, 500 μm, 100 μm, 1 μm, 500 nm, 400 nm, 300 nm, 200 nm, or 100 nm. In some embodiments, the plurality of micro-scale patterns or nano-scale patterns, or a combination thereof, are made by mechanical embossing at room temperature.

[0224] Functionalizable layer

[0225] In any of the disclosed embodiments, the functionalizable layer may include a reactive silane layer, a functionalizable hydrogel, or a functionalizable polymer. In some such embodiments, the functionalizable layer may include one or more functional groups. In some such embodiments, the functional group may be selected from: optionally substituted olefins, azides, optionally substituted amines, carboxylic acids, optionally substituted hydrazones, halogens, hydroxyls, optionally substituted tetrazoles, optionally substituted tetrazines, thiols, and combinations thereof. In some further embodiments, the functionalizable layer comprises a polymer or hydrogel comprising formula (Ia) or (Ib):

[0226]

[0227] R 1 is H or optionally substituted alkyl; the functional group R A is selected from the group consisting of: azide, optionally substituted amine, optionally substituted olefin, optionally substituted hydrazone, carboxylic acid, halogen, hydroxyl, optionally substituted tetrazole, optionally substituted tetrazine, and thiol; R 5 is selected from H or optionally substituted alkyl; each -(CH2)-p may be optionally substituted; p is an integer from 1 to 50; n is an integer from 1 to 50,000; and m is an integer from 1 to 100,000. In some such embodiments, the functional group comprises azide. In some embodiments, each R 1 and each R 5 is hydrogen. In some embodiments, the functional group R Ais an azide. In some embodiments, p is 5. In one embodiment, the polymer or hydrogel contained in the functionalizable layer is PAZAM. Methods for preparing and using PAZAM, and other functionalizable materials that can be used in layers of substrates of the present disclosure, are described in U.S. Patent Application Nos. 13 / 784,368 and 61 / 841,647.

[0228] Examples of reactive silanes that can be used include, but are not limited to: acrylate functional silanes, aldehyde functional silanes, amino functional silanes, anhydride functional silanes, azide functional silanes, carboxylate functional silanes, phosphonate functional silanes, sulfonate functional silanes, epoxy functional silanes, ester functional silanes, vinyl functional silanes, olefin functional silanes, halogen functional silanes, and dipodal silanes with or without any of the above functional groups. Norbornene silanes are particularly useful and are described in, for example, U.S. Patent Application No. 61 / 841,647. The choice of silane functionality can be made based on the reactivity of the material to be reacted with the silane. For example, amino functional silanes react with thermoplastics such as polyacrylates, polyamides, polyamide-imides, polybutylene terephthalates, polycarbonates, polyether ketones, polyethylene, polyphenylene sulfide, polysulfones, polyvinyl butyral, and polyvinyl chloride. Vinyl-functional silanes and olefin-functional silanes react with thermoplastics such as polyacetal, polyethylene, and polypropylene. Acrylate-functional silanes react with thermoplastics such as polypropylene and polystyrene.

[0229] In any disclosed embodiment, the functional group of the functionalizable layer is connected to a biomolecule. In some embodiments, the biomolecule is selected from: amino acids, nucleosides, nucleotides, peptides, oligonucleotides, polynucleotides, nucleic acids, proteins, or combinations thereof. In some such embodiments, the biomolecule is a nucleic acid.

[0230] Functionalizable layer - hydrogel

[0231] In some embodiments, the functionalizable layer can include a hydrogel. Non-limiting examples of hydrogels that can be used in the present application are described herein.

[0232] WO 00 / 31148 discloses polyacrylamide hydrogels and arrays based on polyacrylamide hydrogels, in which a so-called polyacrylamide prepolymer is preferably formed from acrylamide and acrylic acid or an acrylic acid derivative containing vinyl groups. Crosslinking of the prepolymer may then take effect. The hydrogel thus produced is immobilized, preferably on glass. Functionalization of the immobilized hydrogel can also be achieved.

[0233] WO 01 / 01143 describes a technology similar to WO 00 / 31148, but differs in that the hydrogel has functionality capable of participating in a [2+2] photocycloaddition reaction with biomolecules to form fixed arrays of such biomolecules. The functionality of dimethylmaleimide (DMI) is particularly preferred. The use of [2+2] photocycloaddition reactions in the context of polyacrylamide-based microarray technology is also described in WO02 / 12566 and WO03 / 014392.

[0234] US Patent No. 6,465,178 discloses the use of a reagent composition for providing an activated slide for preparing a microarray of nucleic acids; the reagent composition includes an acrylamide copolymer.

[0235] WO 00 / 53812 discloses the preparation of polyacrylamide-based DNA hydrogel arrays and the use of these arrays in replicative amplification.

[0236] If desired, once the hydrogels are formed, biomolecules can then be attached to them to produce molecular arrays. Attachment has been achieved in different ways. For example, U.S. Pat. No. 6,372,813 teaches the immobilization of polynucleotides having dimethylmaleimide groups to a hydrogel produced having dimethylmaleimide groups, wherein the immobilization is obtained by performing a [2+2] photocycloaddition step between two dimethylmaleimide groups (one attached to the polynucleotide to be solidified and one pendant to the hydrogel).

[0237] When the molecular array is formed after the hydrogel is produced, two strategies can be used to achieve this goal. First, the hydrogel can be chemically modified after it is produced. A more common alternative is to achieve polymerization by reacting the molecules to be arrayed with comonomers with prepared or preactivated functionality.

[0238] Alternatives to initially forming a hydrogel and subsequently arranging molecules thereto have been described, for example, forming an array while the hydrogel is being produced. This can be accomplished, for example, by direct copolymerization of acrylamide-derived polynucleotides. An example of this approach is described in WO01 / 62982, where acrylamide-derived polynucleotides are mixed with an acrylamide solution and polymerization is accomplished directly.

[0239] Mosaic Technologies (Boston, Mass., USA) produces ACRYDITE TM (an acrylamide phosphoramidite), which can be reacted with the polynucleotide before the resulting monomer is copolymerized with acrylamide.

[0240] Efimov et al. (Nucleic Acids Research, 1999, 27(22), 4416-4426) disclose additional examples of simultaneous hydrogel / array formation in which copolymerization of acrylamide, reactive acrylic acid derivatives, and modified polynucleotides having 5'- or 3'-terminal acrylamide groups plays a role.

[0241] The compositions and methods described above and in the references cited above can be used in the compositions and methods described herein.For example, a hydrogel can be used as a functionalizable layer in a substrate described herein.

[0242] Functionalizable layer - polymer

[0243] In some embodiments, the functionalizable layer can include a polymer having one or more functional groups capable of reacting with a target biomolecule. In some such embodiments, the functional group can be selected from: optionally substituted olefins, azides, optionally substituted amines, carboxylic acids, optionally substituted hydrazones, halogens, hydroxyls, optionally substituted tetrazoles, optionally substituted tetrazines, thiols, and combinations thereof. Non-limiting examples of polymers useful for the present application are described herein, including those described in U.S. Serial Nos. 13 / 784,368 and 2011 / 0059865, which are incorporated herein by reference in their entirety.

[0244] PAZAM

[0245] In some embodiments, the polymer of formula (Ia) or (Ib) is also represented by formula (IIa) or (IIb):

[0246]

[0247] wherein n is an integer from 1 to 20,000, and m is an integer from 1 to 100,000.

[0248] In some embodiments, the functionalized molecule used for direct conjugation is poly(N-(5-azidoacetamidopentyl)acrylamide-co-acrylamide) (PAZAM). PAZAM can be prepared by polymerization of acrylamide and Azapa (N-(5-(2-azidoacetamido)pentyl)acrylamide) in any ratio. In some embodiments, PAZAM is a linear polymer. In some other embodiments, PAZAM is a lightly cross-linked polymer. In some embodiments, PAZAM is applied as an aqueous solution. In some other embodiments, PAZAM is applied as an aqueous solution containing one or more solvent additives such as ethanol. Methods for preparing different PAZAM polymers are discussed in detail in U.S. Serial No. 13 / 784,368, which is incorporated herein by reference in its entirety.

[0249] Polymer layer - photocurable polymer

[0250] In any disclosed embodiment, the polymer layer may include at least one photocurable polymer. In some embodiments, the photocurable polymer includes urethane, acrylate, siloxane, epoxy, polyacrylic acid, polyacrylate, epoxysiloxane, epoxy resin, polydimethylsiloxane (PDMS), silsesquioxane, acyloxysilane, maleic acid polyester, vinyl ether, monomers containing vinyl or ethynyl groups, or copolymers and combinations thereof.

[0251] In any disclosed embodiment, the thickness of the backing layer can be, for example, no more than about 10 nm, 50 nm, 100 nm, 1 μm, 10 μm, 100 μm, 500 μm, 1 mm, 5 mm, 1 cm, or 10 cm. Alternatively or additionally, the thickness of the backing layer is not less than about 10 nm, 50 nm, 100 nm, 1 μm, 10 μm, 100 μm, 500 μm, 1 mm, 5 mm, 1 cm, or 10 cm.

[0252] In any disclosed embodiment, the thickness of the polymer layer is no more than about the length of a carbon-carbon bond, 1 nm, 10 nm, 50 nm, 100 nm, 1 μm, 10 μm, 100 μm, 500 μm, 1 mm, 5 mm, 1 cm, or 10 cm. In some embodiments, the thickness of the polymer layer is no less than about 10 nm, 50 nm, 100 nm, 1 μm, 10 μm, 100 μm, 500 μm, 1 mm, 5 mm, 1 cm, or 10 cm.

[0253] In any disclosed embodiment, the thickness of the functionalizable layer is no more than about the length of a carbon-carbon bond, 1 nm, 10 nm, 50 nm, 100 nm, 1 μm, 10 μm, 100 μm, 500 μm, 1 mm, 5 mm, 1 cm, or 10 cm. In some embodiments, the thickness of the functionalizable layer is no less than about 10 nm, 50 nm, 100 nm, 1 μm, 10 μm, 100 μm, 500 μm, 1 mm, 5 mm, 1 cm, or 10 cm.

[0254] Exemplary dicing tapes that can be used in the compositions or methods described herein include, but are not limited to: Lintec KL-3225 UV tape; Ultron Systems, antistatic UV adhesive plastic film (p / n: 1043R-9.05), or AITU VR250 (AI Technology Inc., Princeton Junction, NJ). Other useful dicing tapes are commercially available from Semiconductor Equipment Corp. (Moorpark, CA), such as low viscosity, expandable PVC, 80um thick (P / N 24213); extra high viscosity, non-expandable polyethylene, 90um thick (P / N 24216); ultra-high viscosity, expandable polyolefin, 85um thick (P / N 24339); or ultra-high viscosity, expandable polyolefin, 170um thick (P / N 24351). The UC series of UV tapes for wafer dicing sold by Furukawa Electric Co., Ltd (Tokyo, Japan) are also useful.

[0255] Backing layer

[0256] In any disclosed embodiment, the backing layer can be made of a material selected from the group consisting of silicon dioxide, plastics, quartz, metals, metal oxides, paper and combinations thereof. In some such embodiments, the backing layer is made of a flexible plastic material. In some such embodiments, when the backing layer is contacted with a solid support in the method described herein, the flexible plastic material has the hardness that is enough to prevent the backing layer from conforming to the nanometer-level profile or micrometer-level profile on the solid support. For example, under sufficient pressure, contact between the solid support and the backing layer may occur so that the negative replica of the nanometer-level profile or micrometer-level profile is transferred to the adhesive layer connected with the backing layer; However, the backing layer will have enough hardness to resist deformation for a long time or even temporarily resist deformation in some cases. In some such embodiments, the backing layer is made of a roll of flexible film. The non-limiting examples of the backing layer that can be used for the application are described herein.

[0257] In some embodiments, the backing layer for the present application includes materials based on silicon dioxide, for example: glass, fused silica and other materials containing silicon dioxide. In some embodiments, the substrate based on silicon dioxide can also be silicon, silicon dioxide, silicon nitride, silane (silicone hydrides). In some embodiments, the backing layer for the present application includes plastic materials such as: polyethylene, polystyrene, poly (vinyl chloride), polypropylene, nylon, polyester, polycarbonate and poly (methyl methacrylate). Preferred plastic materials are poly (methyl methacrylate), polystyrene and cycloolefin polymer substrates.

[0258] In some other embodiments, the backing layer can be a metal. In some such embodiments, the metal is gold. In some embodiments, the backing layer has at least one surface comprising a metal oxide. In one embodiment, the surface comprises tantalum oxide.

[0259] Acrylamide, enone or acrylate can also be used as backing layer materials. Other backing layer materials can include but are not limited to: gallium arsenide, indium phosphide, aluminum, ceramics, polyimide, quartz, resins, polymers and copolymers. The above list is intended to be illustrative, but not limited to this application.

[0260] In some embodiments, the backing layer and / or the backing layer surface can be quartz. In some other embodiments, the backing layer and / or the backing layer surface can be a semiconductor, ie, GaAs or ITO.

[0261] In some embodiments, the backing layer is made of paper.

[0262] In some embodiments, the backing layer is made of a flexible material. In some such embodiments, the backing layer is made of a roll of flexible film (eg, having a thickness of less than about 1 mm, 500 μm, 100 μm, or less).

[0263] The backing layer may comprise a single material or a plurality of different materials, for example, two or more layers of one or more materials.The shape of the backing layer used may vary depending on the application being implemented.

[0264] Doping layer

[0265] One or more layers present in the compositions described herein or used in the methods described herein may be doped with materials having desired properties. For example, the layer may have plasma properties due to the presence of nanoparticles, quantum dots, or the like incorporated into the material of the layer. The resulting plasma properties may provide an antenna effect during a subsequent detection step in the doped layer or on the doped layer. Alternatively or additionally, the layer may be doped with a receptor, ligand, or nucleic acid to provide binding specificity in analytical detection applications; or doped with reactive moieties such as those described in the context of the functionalizable layer to provide subsequent functionalization; or doped with a material that changes the optical properties of the layer (such as a filter material), a material that absorbs light in a specific wavelength range, a material that emits light in a specific wavelength range, or a material that scatters light; or doped with a material that imparts conductive properties such as conductivity; or doped with a thermally conductive material.

[0266] In a specific embodiment, the layer can be doped before being used in the method described herein. For example, one or more layers of the substrate can be doped before the pattern is generated in the substrate. By way of a more specific example, the polymer layer or the functionalizable layer can be doped before the pattern is generated in the polymer layer. In this way, the dopant material in the polymer layer will form a pattern portion, displacing it from some areas of the pattern and retaining it in other areas of the pattern. The dopant material in the functionalizable layer can be exposed to some parts of the pattern and protected in other parts of the pattern to allow selective detection or selective reactivity through the exposed parts in the subsequent use of the substrate.

[0267] One or more layers described herein may be doped, including, for example, a polymer layer, a functionalizable layer, or a backing layer. The dopant material may be present primarily or only on the surface of the layer, within the volume of the layer, or both within the layer and on the surface of the layer.

[0268] Substrate Application

[0269] In some embodiments, substrate as described herein forms at least a portion of a flow cell or is located in a flow cell. In some such embodiments, the flow cell also includes polynucleotides, which are connected to the surface of the substrate by reacting with the functional group of a functionalizable layer (e.g., a hydrogel coating). In some embodiments, the functionalizable layer is arranged between a polymer layer and a backing layer, wherein at least a portion of the micrometer-scale pattern or nanometer-scale pattern of the polymer layer is penetrated to expose the functionalizable layer below. In some embodiments, the polymer layer is arranged between a functionalizable layer and a backing layer, wherein at least a portion of the micrometer-scale pattern or nanometer-scale pattern of the functionalizable layer is penetrated to expose the polymer layer below.

[0270] In some embodiments, the polynucleotides are present in a flow cell in a polynucleotide cluster, wherein the polynucleotides of the polynucleotide cluster are attached to the surface of the flow cell by a hydrogel coating. The clusters can be formed by solid phase amplification techniques such as solid phase PCR, bridge amplification, or other techniques known in the art or described in the following: U.S. Patent No. 5,641,658; U.S. Patent No. 2002 / 0055100; U.S. Patent No. 7,115,400; U.S. Patent No. 2004 / 0096853; U.S. Patent No. 2004 / 0002090; U.S. Patent No. 2007 / 0128624; and U.S. Patent No. 2008 / 0009420, each of which is incorporated herein by reference. In a preferred embodiment, the flow cell is a flow chamber divided into a plurality of lanes or sectors, wherein one or more of the plurality of lanes or sectors comprises a surface coated with a covalently attached functionalizable hydrogel coating as described herein. In some embodiments of the flow cell described herein, the polynucleotides connected in a single polynucleotide cluster have identical or similar nucleotide sequences. In some embodiments of the flow cell described herein, the connected polynucleotides of different polynucleotide clusters have different or non-similar nucleotide sequences. Exemplary flow cells that can be used in the methods and compositions described herein and substrates for making the flow cell include, but are not limited to, commercially available from Illumina, Inc. (San Diego, CA) or described in US 2010 / 0111768 A1 or US 2012 / 0270305, each of which is incorporated herein by reference.

[0271] In some embodiments, the substrate described herein forms at least a portion of a microfluidic device or is located in a microfluidic device. Exemplary devices include flow cells for nucleic acid sequencing and other analytical procedures, such as those commercially available from Illumina, Inc. (San Diego, CA) or described in US2010 / 0111768 A1 or US2012 / 0270305 A1, each of which is incorporated herein by reference. Other microfluidic devices including cell sorting devices, chip lab devices, capillary sequencers, cell counting devices, etc. can also be prepared at least in part by substrates described herein or include substrates described herein.

[0272] Substrate preparation

[0273] Some embodiments disclosed herein include methods for preparing a substrate, comprising: providing a substrate comprising a functionalizable layer comprising one or more functional groups, wherein the functionalizable layer is disposed between a backing layer and a photocurable polymer layer; contacting a surface of the photocurable polymer layer with a template containing a plurality of micrometer-scale patterns or nanometer-scale patterns including micrometer-scale or nanometer-scale holes, pillars, channels, intersections, or a combination thereof; applying pressure to the template or substrate to transfer the micrometer-scale pattern or nanometer-scale pattern to at least the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature; irradiating the photocurable polymer with UV light to cure the photocurable polymer; and separating the template from the substrate; wherein at least a portion of the polymer layer is penetrated to expose the functionalizable layer underneath.

[0274] Figures 8A-8D A cross-sectional view of a substrate 800 according to an embodiment of the present invention prepared by a UV nanoembossing method is shown.

[0275] exist Fig. 8A In the embodiment, a plurality of nanoscale patterns 805 and 806 are included on an embossing stamp 804, and a plurality of grooves 807 are included between the nanoscale patterns. A substrate 800 including a backing layer 801, a functionalizable layer 802, and a photocurable polymer layer 803a is also provided.

[0276] Subsequently, the embossing stamp 804 is brought into contact with the upper surface of the photocurable polymer layer 803a, and then Figure 8B As shown in FIG. 8 , pressure is applied to an embossing stamp 804 or a substrate 800. Nanoscale patterns 805 and 806 are formed in a photocurable polymer layer 803a of the substrate 800.

[0277] Then, if Figure 8C As shown, UV light is irradiated on the UV curable polymer layer 803a through an embossing stamp 804. The embossing stamp 804 is made of a material that allows UV light to pass through, so that the UV light reaches the photocurable polymer layer 803a and cures the photocurable polymer to form a polymer layer 803. Alternatively, if the backing layer and the functionalizable layer 802 are made of a material that allows UV light to pass through, the UV light can be applied through the backing layer 801.

[0278] Then, if Fig.8D As shown, embossing stamp 804 is separated from polymer layer 803, resulting in the formation of nanoscale patterns 805 and 806 on substrate 800. Nanoscale pattern 805 penetrates polymer layer 803 to expose functionalizable layer 802 underneath.

[0279] Some embodiments disclosed herein include methods for preparing a substrate, comprising: providing a template comprising a plurality of micrometer-scale patterns or nanometer-scale patterns comprising micrometer-scale or nanometer-scale holes, channels, or a combination thereof on the surface of the template; depositing a functional layer comprising one or more functional groups on the surface of the template so that at least a portion of the micrometer-scale or nanometer-scale holes, channels, or a combination thereof comprises the functional layer; removing excess functionalizable layer from the surface of the template so that the functionalizable layer is present only on a portion of the surface of the template; providing a substrate comprising a backing layer, wherein the backing layer has a photocurable polymer layer disposed on the backing layer; contacting the surface of the photocurable polymer layer with the surface of the template, wherein the template has a plurality of micrometer-scale patterns or nanometer-scale patterns and the functionalizable layer; applying pressure to the template or substrate to transfer the micrometer-scale patterns or nanometer-scale patterns to at least the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature; irradiating the photocurable polymer with UV light to cure the photocurable polymer; and separating the template from the substrate; wherein at least a portion of the functionalizable layer is transferred to the polymer layer.

[0280] Figures 9A-9E A cross-sectional view of a substrate 900 according to an embodiment of the present invention prepared by a UV nanoembossing method is shown.

[0281] exist Fig. 9A In the embodiment, a plurality of nanoscale patterns 905 are included on an embossing stamp 904, and a plurality of grooves 906 are included between the nanoscale patterns 905. A functionalizable layer 902 is deposited on the nanoscale patterned surface of the embossing stamp 904.

[0282] Later, in Fig. 9B In the embodiment of the present invention, excess functionalizable layer 902 is removed so that the functionalizable layer is present only within the nanoscale pattern 905 of the embossing stamp 904. A substrate 900a having a backing layer 901 and a photocurable polymer layer 903a is provided.

[0283] Then, if Fig. 9C As shown in FIG. 1 , an embossing stamp 904 is brought into contact with the upper surface of the photocurable polymer layer 903a, and then pressure is applied to the embossing stamp 904 or the substrate 900a. A nanoscale pattern 905 is formed in the photocurable polymer layer 903a of the substrate 900a.

[0284] Then, if Fig.9D As shown, UV light is irradiated on the UV curable polymer layer 903a through an embossing stamp 904. The embossing stamp 904 is made of a material that allows UV light to pass through, so that the UV light reaches the photocurable polymer layer 903a and cures the photocurable polymer to form a polymer layer 903. Alternatively, if the backing layer 901 is made of a material that allows UV light to pass through, the UV light can be applied through the backing layer 901.

[0285] Finally, the embossing stamp 904 is separated from the polymer layer 903. The functionalizable layer 902 is transferred to the upper surface of the polymer layer 903. Fig.9E As shown, nanoscale pattern 905 is formed in polymer layer 903 .

[0286] In some embodiments, the embossing stamp may have a micro-scale pattern or a nano-scale pattern that causes at least a portion of the polymeric layer and the functionalizable layer to be penetrated to expose the underlying backing layer.

[0287] In some embodiments, the substrate further comprises a backing layer having a photocurable polymer layer disposed on the backing layer, wherein the substrate is a roll of flexible dicing tape.

[0288] In any of the disclosed method embodiments, the method may further include applying a sealing layer to the substrate after removing the template to substantially seal the polymeric layer and the functionalizable layer between the backing layer and the sealing layer.

[0289] In some embodiments, the sealing layer further comprises a second substrate prepared by the method of any disclosed embodiment, wherein the functionalizable layer and the photocurable polymer layer of the substrate are disposed between the backing layer of the substrate. In some such embodiments, the sealing layer is optically transparent.

[0290] In any disclosed method embodiment, the method can also include forming the substrate into a cylinder. In some embodiments, the backing layer is closer to the outer surface of the cylinder than the functionalized layer or polymer layer of the substrate. In some other embodiments, the backing layer is closer to the inner surface of the cylinder than the functionalized layer or polymer layer of the substrate. In some embodiments, the substrate of cylindrical shape can be formed by using a flexible backing layer and winding the substrate around a cylinder such as a glass rod or a plastic rod or a capillary.

[0291] In any of the disclosed process embodiments, the process can be a roll-to-roll process.

[0292] It should be understood from the above that various embodiments of the present disclosure have been described herein for illustrative purposes, and various modifications may be made without departing from the scope and spirit of the present disclosure. Therefore, the various embodiments disclosed herein are not intended to be limiting, wherein the actual scope and spirit are defined by the claims. Example

[0293] Further embodiments are disclosed in more detail in the following examples, which are not intended in any way to limit the scope of the claims.

[0294] Example 1

[0295] High-fidelity nanoscale pattern transfer

[0296] An 8" master template wafer containing 12 Hi-seq sized patterned flow cells was used for direct embossing into a UV-curable adhesive coated polymer sheet ( Fig. 10A ). A template wafer with a nanoscale patterned structure is aligned with a polymer sheet coated with a UV-curable adhesive layer so that the adhesive layer is in direct contact with the nano-patterned surface. A mechanical force is applied to the non-adhesive surface of the polymer sheet. The polymer sheet is then irradiated with UV light to cure the adhesive and the polymer sheet is separated by delamination. As a result, a high-fidelity pattern transfer from the template to the polymer sheet is observed. The diffraction pattern from the flow cell area confirms the pattern transfer over a large area.

[0297] Flow cell 7 (FC7) was cut out from the polymer sheet for further microscopic examination and electron microscopy. Fig. 10B As shown, dark field microscope images of the fiducial from FC7 demonstrate nanoscale pattern transfer. In this case, the master template contains 350 nm nanopores characterized by a 750 nm pitch. The pattern transferred to the polymer sheet is of opposite polarity to the pattern on the template (nanoramids). In addition, SEM images of the fiducial area of ​​FC7 are shown at 1240X and 20kX magnifications. Fig. 10C and 10D In addition, nanoscale pillars on the polymer sheet are shown at 40kX and 80kX magnifications. Fig.10E and 10F In all cases excellent pattern transfer can be seen with a pillar diameter of about 330 nm and a pillar height of about 300 nm.

[0298] Example 2

[0299] High-fidelity reverse polarity nanoscale pattern transfer

[0300] The polarity of the nanoscale pattern of the master template is opposite compared to the master template described in Example 1. The master template contains nanoscale pillars instead of nanopore features. The same pattern transfer method as described in Example 1 is used to produce a pattern of opposite polarity on the polymer sheet. The formation of nanopores and nanogrooves in the polymer sheet after physical embossing and UV curing is shown in different magnifications. Fig.11A and 11B . High-fidelity and high-throughput pattern transfer was achieved.

[0301] Example 3

[0302] High-throughput embossing of microfluidic structures

[0303] A silicon master template containing microfluidic structures in a patterned SU-8 photoresist was prepared. The silicon master wafer was then embossed into a transparent polymer sheet with a UV-curable adhesive layer so that the microfluidic pattern was in direct contact with the adhesive layer. After standard UV irradiation and separation procedures as described in Example 1, multiple 4-inch polymer replicas of the master wafer were formed in less than 2 minutes. The transferred patterned microfluidic structures are shown in Figures 12A-12D These fluid structures were formed in commercially available dicing tape (Lintec KL-3225 UV tape or Ultron Systems, Anti-Static UV Adhesive Plastic Film p / n: 1043R-9.05).

[0304] Fig. 12A Channels and intersections of serpentine fluids formed for applications such as fluid mixing are shown. Similarly, microfluidic filters ( Fig. 12B ). In addition, the microfluidic structure was tested by using a capillary-driven fluid (1 M NaCl containing a red dye) flowing through a serpentine microfluidic channel. Fig. 12C Optical images of fluid flowing through a microfluidic channel at different time points (t=5 s and t=10 s) are shown. Fig.12D An optical image of fluid flow into a T-junction for fluid mixing is shown.

Claims

1. A roll-to-roll method for preparing a patterned substrate, comprising the following steps: contacting a template comprising a plurality of micrometer-scale or nanometer-scale patterns with a surface of a substrate having a photocurable polymer layer disposed thereon; applying pressure to the template or the substrate to transfer the microscale or nanoscale pattern to the photocurable polymer layer, wherein the contacting and applying pressure are performed at room temperature; irradiating the photocurable polymer of the photocurable polymer layer with UV light to cure the photocurable polymer to form a patterned UV-curable polymer layer; and separating the template from the substrate, The substrate further comprises a functionalizable layer in contact with the photocurable polymer layer, wherein the functionalizable layer comprises a functionalizable polymer, and wherein the functionalizable polymer comprises one or more functional groups capable of being linked to a biomolecule, wherein the biomolecule is an amino acid, a nucleoside, a nucleotide, a peptide, a nucleic acid, a protein, or a combination thereof. 2 . The roll-to-roll method of claim 1 , wherein at least a portion of the microscale or nanoscale pattern of the template is transferred to the functionalizable layer.

3. The roll-to-roll process of claim 1 , wherein the functionalizable layer comprises a polymer of formula (Ia): R 1 is H or optionally substituted alkyl; R A is an azido group, an optionally substituted amino group, an optionally substituted alkenyl group, an optionally substituted hydrazone group, a carboxyl group (—COOH), a halogen group, a hydroxyl group, an optionally substituted tetrazolyl group, an optionally substituted tetrazinyl group, or a thiol group (—SH); R 5 is H or optionally substituted alkyl; p is an integer from 1 to 50; n is an integer from 1 to 50,000; and m is an integer from 1 to 100,000.

4. The roll-to-roll method according to claim 3, wherein R A It is an azido group.

5. The roll-to-roll process of claim 4, wherein the functionalizable layer comprises poly(N-(5-azidoacetamidopentyl)acrylamide-co-acrylamide) (PAZAM).

6. The roll-to-roll method according to any one of claims 1 to 5, wherein the functionalizable layer further comprises a reactive silane layer.

7. The roll-to-roll method according to any one of claims 1 to 5, wherein the biomolecule is an oligonucleotide or a polynucleotide or a combination thereof.

8. The roll-to-roll method according to any one of claims 1 to 5, wherein the photocurable polymer comprises polyurethane, polyacrylate, silicone, polyacrylic acid, epoxy, polyacyloxysilane, maleic polyester, polyvinyl ether, copolymers of the above polymers, or combinations thereof.

9. The roll-to-roll method according to any one of claims 1 to 5, wherein the photocurable polymer comprises epoxysilicone, polydimethylsiloxane or polysilsesquioxane.

10. A roll-to-roll method according to any one of claims 1 to 5, wherein at least one of the photocurable polymer layer or the functionalizable layer further comprises a dopant material, wherein the dopant material is selected from: nanoparticles, nucleic acids, filter materials, light absorbing materials, luminescent materials, light scattering materials, conductive materials, thermally conductive materials and combinations thereof.

11. The roll-to-roll method of claim 10, wherein the dopant material comprises quantum dots.

12. A patterned substrate prepared by the roll-to-roll method of any one of claims 1 to 11.

13. The patterned substrate of claim 12, wherein the micro-scale or nano-scale pattern in the UV-cured polymer layer is capable of allowing access to a sample fluid.

14. The patterned substrate of claim 12, further comprising a fluid reservoir. 15 . The patterned substrate according to claim 12 , wherein at least a portion of the microscale or nanoscale pattern is a columnar body having an average diameter of less than 500 nm.

16. The patterned substrate according to any one of claims 12 to 14, wherein the patterned substrate comprises a backing layer made from a roll of flexible film.

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