Resin composition and flow cell incorporating the same

Abstract

An example resin composition includes an epoxy resin matrix, a first photoacid generator, and a second photoacid generator. The first photoacid generator includes an anion having a molecular weight less than about 250 g / mol. The second photoacid generator includes an anion having a molecular weight greater than about 300 g / mol. In one example, i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cation of the first photoacid generator and the cation of the second photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application Serial No. 62 / 952,821, filed December 23, 2019, the contents of which are incorporated herein by reference in their entirety. Background Art

[0003] Nanoimprint lithography enables the economical and efficient production of nanostructures. Nanoimprint lithography employs direct mechanical deformation of a resist material using a stamp bearing the nanostructures. The resist material solidifies while holding the stamp in place, locking the shape of the nanostructures in the resist.

[0004] Nanoimprint lithography has been used to make patterned substrates, which can be used in various applications. Some patterned substrates include fluid channels and discrete holes or recessed portions. These patterned substrates can be built into flow cells. In some flow cells, active surface chemicals are introduced into discrete recessed portions, and the gap region surrounding the discrete recessed portions keeps inertia. These flow cells may be particularly useful for detecting and assessing various molecules (for example, DNA), molecular families, gene expression levels or single nucleotide polymorphisms. Summary of the Invention

[0005] Disclosed herein are examples and aspects of resin compositions. The resin compositions are suitable for nanoimprint lithography and for producing patterned flow cell surfaces. The resin compositions comprise a specific combination of photoacid generators (PAGs) that, when used together, exhibit surprising synergy that significantly and desirably alters the degree of cure, for example, increasing resin hardness and / or reducing the optical density at 2990 cm-1 over a relatively short period of time. -1 The correction intensity at .

[0006] In a first aspect, a resin composition comprises an epoxy resin matrix; a first photoacid generator comprising an anion having a molecular weight less than 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight greater than about 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light used to cure the resin composition.

[0007] In examples of the first aspect, the epoxy resin matrix comprises an epoxy material selected from the group consisting of: epoxy-functionalized silsesquioxane; trimethylolpropane triglycidyl ether; tetra(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4- epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexylmethyl-3,4-epoxycyclohexane carboxylate; bis((3,4- epoxycyclohexyl)methyl)adipate; 4-vinyl-1-cyclohexene 1,2-epoxide; vinylcyclohexene dioxide; 4,5-epoxytetrahydrophthalic acid diglycidyl ester; 1,2-epoxy-3-phenoxypropane; glycidyl methacrylate; 1,2-epoxyhexadecane; poly(ethylene glycol) diglycidyl ether; pentaerythritol glycidyl ether; cyclohexane-1,2-dicarboxylic acid diglycidyl ester; tetrahydrophthalic acid diglycidyl ester; and combinations thereof.

[0008] In examples of the first aspect, the first photoacid generator is selected from the group consisting of: bis-(4-methylphenyl) iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate.

[0009] In examples of the first aspect, the second photoacid generator is selected from the group consisting of: tetra(perfluorophenyl)borate triphenylsulfonium salt, tetra(pentafluorophenyl)gallic acid salt, and tris[(trifluoromethyl)sulfonyl]methanide.

[0010] In examples of the first aspect, the cation of the first photoacid generator, or the cation of the second photoacid generator, or the cation of the first photoacid generator and the second photoacid generator is selected from the group consisting of: diaryliodonium cations and triphenylsulfonium cations having a maximum absorption wavelength (λ 最大 ) of about 350 nm.

[0011] In examples of the first aspect, the cation of the second photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition; and the cation of the first photoacid generator has a mass attenuation coefficient of less than 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition. In one example, the cation of the second photoacid generator is selected from the group consisting of: diaryliodonium cations and triphenylsulfonium cations having a maximum absorption wavelength (λ 最大) is a diaryliodonium cation and a triphenylsulfonium cation with a wavelength of about 350 nm; and the cation of the first photoacid generator is selected from the group consisting of: a bis-(4-methylphenyl)iodonium cation, a (isopropylbenzene)cyclopentadienylferron (II) cation, a ferrocenium cation, a 1-naphthyldiphenylsulfonium cation, a (4-phenylthienyl)diphenylsulfonium cation, a bis(2,4,6-trimethylphenyl)iodonium cation, a bis(4-tert-butylphenyl)iodonium cation, an N-hydroxy-5-norbornene-2,3-dicarboximide perfluoro-1-butanesulfonate cation and a triarylsulfonium cation.

[0012] In an example of the first aspect, the cation of the first photoacid generator and the cation of the second photoacid generator have respective mass attenuation coefficients of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition.

[0013] In an example of the first aspect, the first photoacid generator is present in an amount ranging from about 1% to about 5% by mass of the total solids in the resin composition; and the second photoacid generator is present in an amount ranging from about 0.1% to about 2% by mass of the total solids in the resin composition.

[0014] In an example of the first aspect, the molecular weight of the anion of the second photoacid generator is in a range from greater than about 300 g / mol to about 1,000 g / mol.

[0015] It should be understood that any features of the resin compositions disclosed herein may be combined in any desired manner and / or configuration to achieve the benefits described in this disclosure, including, for example, increased degree of cure.

[0016] In a second aspect, a resin composition comprises an epoxy resin matrix comprising: an epoxy-functionalized polyhedral oligomeric silsesquioxane; a first photoacid generator comprising an anion having a molecular weight of less than about 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight in the range of about 300 g / mol to about 1,000 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light used to cure the resin composition.

[0017] In an example of the second aspect, the epoxy-functionalized polyhedral oligomeric silsesquioxane is selected from the group consisting of a glycidyl-functionalized polyhedral oligomeric silsesquioxane, an epoxycyclohexylethyl-functionalized polyhedral oligomeric silsesquioxane, and combinations thereof.

[0018] In an example of the second aspect, the epoxy resin matrix further comprises an additional epoxy material selected from the group consisting of: trimethylolpropane triglycidyl ether; tetra(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; a copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4-epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexylmethyl-3,4-epoxycyclohexanecarboxylic acid diglycidyl ether; pentaerythritol glycidyl ether; cyclohexane-1,2-dicarboxylate diglycidyl ester; diglycidyl tetrahydrophthalate; and combinations thereof.

[0019] In an example of the second aspect, the first photoacid generator is present in an amount ranging from about 1% to about 5% by mass of the total solids in the resin composition.

[0020] In an example of the second aspect, the second photoacid generator is present in an amount ranging from about 0.1 mass % to about 2 mass % of the total solids in the resin composition.

[0021] In an example of the second aspect, the first photoacid generator is selected from the group consisting of bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate.

[0022] In an example of the second aspect, the second photoacid generator is selected from the group consisting of triphenylsulfonium tetrakis(perfluorophenyl)borate, tetrakis(pentafluorophenyl)gallate, and tris[(trifluoromethyl)sulfonyl]methanate.

[0023] In an example of the second aspect, the cation of the first photoacid generator, or the cation of the second photoacid generator, or both the cations of the first photoacid generator and the second photoacid generator are selected from the group consisting of: maximum absorption wavelength (λ 最大 ) are diaryliodonium cations and triphenylsulfonium cations of about 350 nm.

[0024] In an example of the second aspect, the cation of the second photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition; and the cation of the first photoacid generator has a mass attenuation coefficient of less than 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition. In one example, the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大 ) is a diaryliodonium cation and a triphenylsulfonium cation with a wavelength of about 350 nm; and the cation of the first photoacid generator is selected from the group consisting of: a bis-(4-methylphenyl)iodonium cation, a (isopropylbenzene)cyclopentadienylferron (II) cation, a ferrocenium cation, a 1-naphthyldiphenylsulfonium cation, a (4-phenylthienyl)diphenylsulfonium cation, a bis(2,4,6-trimethylphenyl)iodonium cation, a bis(4-tert-butylphenyl)iodonium cation, an N-hydroxy-5-norbornene-2,3-dicarboximide perfluoro-1-butanesulfonate cation and a triarylsulfonium cation.

[0025] In an example of the second aspect, the cation of the first photoacid generator and the cation of the second photoacid generator have respective mass attenuation coefficients of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition.

[0026] It should be understood that any features of the resin composition disclosed herein can be combined in any desired manner. In addition, it should be understood that any combination of features of the resin composition and / or the first aspect of the resin composition can be used together and / or combined with any examples disclosed herein to achieve the benefits described in the present disclosure, including, for example, increased degree of cure.

[0027] In a third aspect, a flow cell comprises a substrate; and a cured patterned resin on the substrate, the cured patterned resin comprising recessed portions separated by gap regions, and the cured patterned resin comprising a cured form of a resin composition comprising: an epoxy resin matrix; a first photoacid generator comprising an anion having a molecular weight less than about 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight greater than about 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition.

[0028] The example of the third aspect further includes a hydrogel in the recess; and an amplification primer attached to the hydrogel.

[0029] In examples of the third aspect, the epoxy resin matrix comprises an epoxy material selected from the group consisting of: epoxy-functionalized silsesquioxane; trimethylolpropane triglycidyl ether; tetra(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4- epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexylmethyl-3,4-epoxycyclohexane carboxylate; bis((3,4- epoxycyclohexyl)methyl)adipate; 4-vinyl-1-cyclohexene 1,2-epoxide; vinylcyclohexene dioxide; 4,5-epoxytetrahydrophthalic acid diglycidyl ester; 1,2-epoxy-3-phenoxypropane; glycidyl methacrylate; 1,2-epoxyhexadecane; poly(ethylene glycol) diglycidyl ether; pentaerythritol glycidyl ether; cyclohexane-1,2-dicarboxylic acid diglycidyl ester; tetrahydrophthalic acid diglycidyl ester; and combinations thereof.

[0030] In examples of the third aspect, the first photoacid generator is selected from the group consisting of: bis-(4-methylphenyl) iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate; and the second photoacid generator is selected from the group consisting of: tetra(perfluorophenyl)borate triphenylsulfonium salt, tetra(pentafluorophenyl)gallic acid salt, and tris[(trifluoromethyl)sulfonyl]methanide.

[0031] It should be appreciated that any of the features of the flow cell disclosed herein can be combined together in any desired manner. Further, it should be appreciated that any combination of the features of the first aspect of the flow cell and / or the resin composition and / or the second aspect of the resin composition can be used together, and / or in combination with any of the examples disclosed herein to achieve the benefits as described in the present disclosure, including, for example, reduced autofluorescence.

[0032] In a fourth aspect, a method includes depositing a resin composition on a substrate, the resin composition comprising: an epoxy resin matrix; a first photoacid generator comprising an anion having a molecular weight of less than about 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight of greater than about 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition; nanoimprinting the deposited resin composition using a working stamp; and exposing the nanoimprinted deposited resin composition in the incident light for 30 seconds or less at an energy dose ranging from about 0.5 J to about 10 J to form a cured patterned resin.

[0033] It should be appreciated that any feature of the methods disclosed herein can be combined together in any desired manner. Further, it should be appreciated that any combination of features of the first aspect of the method and / or flow cell and / or the second aspect of the resin composition and / or features of the resin composition of this aspect can be used together, and / or combined with any of the examples disclosed herein to achieve the benefits as described in the present disclosure, including, for example, reduced autofluorescence.

[0034] In a fifth aspect, a resin composition comprises an epoxy resin matrix; a first photoacid generator comprising a first anion having a first molecular weight; and a second photoacid generator comprising a second anion having a second molecular weight that is at least 50 g / mol greater than the first molecular weight; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition.

[0035] It should be appreciated that any feature of this aspect of the resin composition disclosed herein can be combined together in any desired manner. Further, it should be appreciated that this aspect of the resin composition and / or any combination of features of the first aspect of the method and / or flow cell and / or the second aspect of the resin composition and / or features of the resin composition of this aspect can be used together, and / or combined with any of the examples disclosed herein to achieve the benefits as described in the present disclosure, including, for example, increased degree of cure. BRIEF DESCRIPTION OF DRAWINGS

[0036] Features of examples of the present disclosure will become apparent by reference to the following detailed description and drawings, of which like reference numerals refer to like parts throughout several views. For the purposes of clarity, not all of the features of the examples are described in detail within this description, but all such features can be understood from this disclosure, and are illustrated in the various drawings. Like reference numerals can have been used throughout the specification and / or drawings to reference like features.

[0037] Figure 1 is a flow chart illustrating examples of the methods disclosed herein;

[0038] Figures 2A to 2E is a schematic perspective view together illustrating examples of the methods disclosed herein;

[0039] Figure 2F is a schematic cross-sectional view taken along line 2F-2F of Figure 2E

[0040] Figure 3 is a graph illustrating corrected infrared (IR) intensity (Y-axis) at 2990 cm -1 versus UV curing time (in seconds, X-axis) for a comparative example resin and one example resin including a combination of photoacid generators as disclosed herein;

[0041] Figure 4 is a graph illustrating corrected IR intensity (Y-axis) at 2990 cm -1 versus UV light dose (in Joules, X-axis) for a comparative example resin and one example resin including a combination of photoacid generators as disclosed herein;

[0042] Figure 5 is a graph illustrating hardness (GPa, left Y-axis) and corrected IR intensity (right Y-axis) at 2990 cm -1 versus indentation number (indentation #, X-axis) for several different indentations prepared from examples of the resins disclosed herein;

[0043] Figure 6 is a graph illustrating autofluorescence (blue intensity, Y-axis) versus UV curing time (in seconds, X-axis) for nine example resins including different combinations of photoacid generators as disclosed herein; and

[0044] Figure 7 is a graph illustrating corrected infrared (IR) intensity (Y-axis) at 2990 cm -1 versus UV curing time (in seconds, X-axis) for nine example resins including different combinations of photoacid generators as disclosed herein. DETAILED DESCRIPTION

[0045] ​Some patterned flow cells include a resin material having discrete holes or recesses formed therein. To form the holes or recesses, various techniques such as nanoimprint lithography can be used to imprint the resin material. Nanoimprint lithography involves imprinting a resin with a template, and then curing the resin. Any features of the template are transferred to the resin.

[0046] Disclosed herein is a resin composition suitable for use in nanoimprint lithography and adapted to produce a patterned flow cell surface. The resin composition includes a particular combination of photoacid generators. As demonstrated in the Examples section set forth herein, when the photoacid generators are used together, they have a surprising synergy that significantly and desirably alters the degree of cure, e.g., increases the resin hardness and / or decreases the corrected intensity at 2990 cm -1 Examples of the resin composition disclosed herein exhibit consistently high degrees of cure in less than 30 seconds. In some cases, a high degree of cure is achieved in less than 5 seconds as compared to other resins that do not include the particular combination of photoacid generators and that have a cure time of 50 seconds, 100 seconds, or more, which is a reduction of at least 90% in cure time.

[0047] The altered degree of cure is combined with the fact that the desired properties of the cured resin, such as hardness and low autofluorescence, are not detrimentally affected by the faster curing process.

[0048] Resin that is not sufficiently cured does not fully vitrify and can exhibit reflow, which can be manifested in poor and uncontrolled hole / recess shape in the patterned area. In addition, resin that is not sufficiently cured can have a low hardness value. Low hardness of the cured resin can increase the susceptibility of the material to downstream processing. For example, low hardness can result in increased scratching during subsequent flow cell manufacturing processes, such as polishing. Since the hardness of examples of the cured resin disclosed herein is in the desired range (e.g., in the range of about 0.22 GPa to about 0.35 GPa, or about 0.25 GPa to about 0.3 GPa), the likelihood of reflow and scratching (or other detrimental downstream processing effects) is minimal or nonexistent.

[0049] Some solidifying resins exhibit undesirable levels of autofluorescence at excitation wavelengths of interest (e.g., violet excitation wavelengths ranging from about 380 nm to about 450 nm, or blue excitation wavelengths ranging from about 450 nm to about 495 nm, or green excitation wavelengths ranging from about 495 nm to about 570 nm). Fluorescence from the solidifying resin can add background noise when imaging the optical labels of nucleotides that have been incorporated into individual nascent strands formed in the recesses during sequencing. Increased background noise can reduce the signal-to-noise ratio (SNR), making it more difficult to resolve signals from individual clusters within individual recesses during sequencing. Examples of the solidifying resin compositions disclosed herein have minimal blue excitation absorbance, resulting in relatively low or no blue or longer wavelength autofluorescence when exposed to violet or blue excitation wavelengths. Thus, the potential for signal interference during sequencing on the patterned flowcell surface of examples of the solidifying resins disclosed herein is minimal or nonexistent.

[0050] Likewise, the resin compositions disclosed herein can have the benefit of significantly improving the throughput of patterned flowcell fabrication without compromising the desired properties of the corresponding solidifying resin.

[0051] definition

[0052] It should be understood that the terms used herein are understood to be given their ordinary meaning in the relevant art unless otherwise indicated. Several terms used herein are listed below with their meanings.

[0053] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0054] The terms include, comprise, contain, and various forms of these terms are all synonymous with each other and are intended to be equally broad.

[0055] The terms top, bottom, lower, upper, and the like are used herein to describe the flowcell and / or various components of the flowcell. It should be understood that these directional terms are not intended to imply a particular orientation, but are used to designate relative orientation between components. The use of directional terms should not be interpreted to limit the examples disclosed herein to any particular orientation.

[0056] It should be understood that the ranges provided herein include the stated range and any value or sub-range within the stated range, as if such value or sub-range were explicitly recited. For example, a range of about 400 nm to about 1 μm (1000 nm) should be interpreted as including not only the explicitly recited limits of about 400 nm to about 1 μm, but also individual values ​​such as about 708 nm, about 945.5 nm, etc., and sub-ranges such as about 425 nm to about 825 nm, about 550 nm to about 940 nm, etc. In addition, when "about" and / or "substantially" are used to describe values, they are intended to cover minor variations (up to + / - 10%) from that value.

[0057] Acrylamide is a functional groups or monomers comprising an acrylamide group. Acrylamide may be a chemical compound acrylamide (e.g., methacrylamide) having a substituent replacing one or more hydrogen atoms. Examples of monomers comprising an acrylamide group include azidoacetamidopentyl acrylamide: and N-isopropylacrylamide: Other acrylamide monomers can be used, some examples of which are described herein.

[0058] As used herein, an "aldehyde" is an organic compound containing a functional group having the structure -CHO, which includes a carbonyl center (i.e., a carbon double-bonded to an oxygen), wherein the carbon atom is also bonded to a hydrogen and an R group, such as an alkyl or other side chain. The general structure of an aldehyde is:

[0059] As used herein, "alkyl" refers to a straight or branched hydrocarbon chain that is fully saturated (i.e., does not contain double and triple bonds). An alkyl group may have from 1 to 20 carbon atoms. Typical alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, and the like. As an example, the designation "C1-C4 alkyl" indicates that there are one to four carbon atoms in the alkyl chain, i.e., the alkyl chain is selected from the group consisting of methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, and tert-butyl.

[0060] As used herein, "alkenyl" refers to a straight or branched hydrocarbon chain containing one or more double bonds. An alkenyl group can have 2 to 20 carbon atoms. Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and the like.

[0061] As used herein, "alkyne" or "alkynyl" refers to a straight or branched hydrocarbon chain containing one or more triple bonds. An alkynyl group can have from 2 to 20 carbon atoms.

[0062] As used herein, "aryl" refers to an aromatic ring or ring system containing only carbon in the ring backbone (i.e., two or more fused rings sharing two adjacent carbon atoms). When an aryl group is a ring system, each ring in the ring system is aromatic. An aryl group can have 6 to 18 carbon atoms. Examples of aryl groups include phenyl, naphthyl, azulenyl, and anthracenyl.

[0063] An "amine" or "amino" functional group refers to a -NR a R b Group, where R a and R b Each independently selected from hydrogen (e.g. ), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 carbocyclyl, C6-C10 aryl, 5-10 membered heteroaryl and 5-10 membered heterocyclyl, as defined herein.

[0064] As used herein, the term "attached" refers to the state in which two things are joined, fastened, adhered, connected, or bound to each other, directly or indirectly. For example, nucleic acids can be attached to polymer hydrogels via covalent or non-covalent bonds. Covalent bonds are characterized by the sharing of electron pairs between atoms. Non-covalent bonds are physical bonds that do not involve the sharing of electron pairs and can include, for example, hydrogen bonds, ionic bonds, van der Waals forces, hydrophilic interactions, and hydrophobic interactions.

[0065] "Autofluorescence" refers to the emission of light by the cured resin when exposed to a blue excitation wavelength. No autofluorescence means that the fluorescence level is below the threshold detection limit. The term "low autofluorescence" refers to the emission level (of the cured resin when exposed to a blue excitation wavelength) being above the threshold detection limit and being low enough to be considered noise, and the noise being at a level that does not interfere with the identification of cluster signals during sequencing (e.g., the autofluorescence level achieves a sufficiently high signal-to-noise ratio (SNR) such that signals from individual clusters can be distinguished during sequencing).

[0066] It should be understood that with respect to quantifying autofluorescence (AF), the definition of "low" or "low level" may vary depending on the tool used to measure the autofluorescence and / or the lamp used to provide the excitation radiation. In some examples, a reference level may be used to define relative AF levels. As an example, the reference level is 0.7 mm thick. The AF level of a solid-state glass (CEXG) is measured, and "low AF" can be defined relative to the CEXG output under blue laser excitation. The value of this output (in arbitrary units) is relevant in a relative sense, as it can depend on the material being measured, the excitation and emission bands being measured, the intensity of the excitation light, etc. As an example, considering that the emission value for a 0.7 mm CEXG is approximately 3500 Au, a layer of approximately 500 nm of cured resin can be considered to have low AF if the total signal from the stack is less than approximately 10,000 AU (e.g., the cured resin contribution is less than 2 times the CEXG contribution), or in other examples, less than approximately 7,000 AU (e.g., the cured resin contribution is less than 1 times the CEXG contribution).

[0067] An "azide" or "azido" functional group refers to a -N3 group.

[0068] As used herein, "carbocyclyl" means a non-aromatic cyclic ring or ring system containing only carbon atoms in the main chain of the ring system. When a carbocyclyl is a ring system, two or more rings can be joined together in a fused, bridged or spiro-connected manner. A carbocyclyl can have any degree of saturation, provided that at least one ring in the ring system is not aromatic. Therefore, carbocyclyl includes cycloalkyl, cycloalkenyl and cycloalkynyl. A carbocyclic group can have 3 to 20 carbon atoms. Examples of carbocycles include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cyclohexenyl, 2,3-dihydro-indene, bicyclo[2.2.2]octyl, adamantyl and spiro[4.4]nonyl.

[0069] As used herein, "cycloalkenyl" or "cycloalkene" refers to a carbocyclic ring or ring system having at least one double bond, wherein no ring in the ring system is aromatic. Examples include cyclohexenyl or cyclohexene and norbornenyl or norbornene. Also as used herein, "heterocycloalkenyl" or "heterocycloalkene" means a carbocyclic ring or ring system having at least one heteroatom in the ring backbone, having at least one double bond, wherein no ring in the ring system is aromatic.

[0070] As used herein, "cycloalkyl" refers to a fully saturated (no double or triple bond) monocyclic or polycyclic hydrocarbon ring system. When composed of two or more rings, these rings can be joined together in a fused manner. The cycloalkyl group can contain 3 to 10 atoms in the ring. In some embodiments, the cycloalkyl group can contain 3 to 8 atoms in the ring. The cycloalkyl group can be unsubstituted or substituted. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl.

[0071] As used herein, "cycloalkynyl" or "cycloalkyne" refers to a carbocyclic ring or ring system having at least one triple bond, wherein no ring in the ring system is aromatic. One example is cyclooctyne. Another example is bicyclononyne. Also as used herein, "heterocycloalkynyl" or "heterocycloalkyne" means a carbocyclic ring or ring system having at least one heteroatom in the ring backbone, having at least one triple bond, wherein no ring in the ring system is aromatic.

[0072] As used herein, the term "deposition" refers to any suitable application technique, which may be manual or automated, and in some cases, results in modification of surface properties. Generally speaking, deposition can be performed using vapor deposition techniques, coating techniques, grafting techniques, and the like. Some specific examples include chemical vapor deposition (CVD), spraying (e.g., ultrasonic spraying), spin coating, thick coating or dip coating, doctor blade coating, whipping dispensing, flow through coating, aerosol printing, screen printing, microcontact printing, inkjet printing, and the like.

[0073] As used herein, the term "recess" or "hole" refers to a discrete concave feature in a patterned resin having a surface opening that is at least partially surrounded by a gap region of the resin. The recess can have any of a variety of shapes at the opening in its surface, including, for example, circular, elliptical, square, polygonal, star-shaped (with any number of vertices), etc. The cross-section of the recess taken orthogonally to the surface can be curved, square, polygonal, hyperbolic, conical, angular, etc. As an example, the recess can be a hole or a groove / trough / valley. The recess can also have more complex structures, such as ridges, step features, etc.

[0074] When used in reference to a collection of items, the term "each" is intended to identify a single item in the collection, but does not necessarily refer to every item in the collection. Exceptions may occur if explicitly disclosed or the context clearly dictates otherwise.

[0075] As used herein, the term "epoxy" refers to

[0076] As used herein, the term "circulation cell" is intended to mean a container having a chamber (e.g., including a flow channel) in which a reaction may be carried out, an inlet for delivering reagents to the chamber, and an outlet for removing reagents from the chamber. In some examples, the chamber enables detection of reactions occurring in the chamber. For example, the chamber may include one or more transparent surfaces that allow optical detection of arrays, optically labeled molecules, etc. at the recessed portion.

[0077] As used herein, a "flow channel" may be an area defined between two bonded or otherwise attached components that can selectively receive a liquid sample. In some examples, the flow channel may be defined between a patterned resin and a cover or between two patterned resins and, thus, may be in fluid communication with one or more recesses defined in the patterned resin.

[0078] As used herein, "heteroaryl" refers to an aromatic ring or ring system (i.e., two or more fused rings that share two adjacent atoms) that contains one or more heteroatoms (i.e., elements other than carbon, including but not limited to nitrogen (N), oxygen (O), and sulfur (S)) in the ring backbone. When the heteroaryl group is a ring system, each ring in the ring system is aromatic. A heteroaryl group can have 5-18 ring members.

[0079] As used herein, "heterocycle" means a non-aromatic cyclic ring or ring system containing at least one heteroatom in the ring backbone. The heterocycles can be joined together in a fused, bridged, or spiro-connected manner. The heterocycle can have any degree of saturation, provided that at least one ring in the ring system is not aromatic. In the ring system, heteroatoms can be present in non-aromatic rings or aromatic rings. The heterocyclic group can have 3 to 20 ring members (i.e., the number of atoms constituting the ring backbone, including carbon atoms and heteroatoms). In some examples, the heteroatom is O, N, or S.

[0080] As used herein, the term "hydrazine" or "hydrazine" refers to a -NHNH2 group.

[0081] As used herein, the term "hydrazone" or "hydrazine" refers to Group, where R a and R b are each independently selected from hydrogen (e.g., ), C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkynyl, C3-C7 carbocycle, C6-C10 aryl, 5-10 membered heteroaryl and 5-10 membered heterocycle, as defined herein.

[0082] As used herein, "hydroxy" refers to an -OH group.

[0083] As used herein, the term "gap region" refers to an area of ​​separated recessed portions on a surface (e.g., of a patterned resin). For example, a gap region may separate one feature of an array from another feature of the array. Two features separated from each other may be discrete, i.e., lack physical contact with each other. In another example, a gap region may separate a first portion of a feature from a second portion of a feature. In many examples, the gap region is continuous, while the features are discrete, e.g., as in the case of a plurality of holes defined in an otherwise continuous surface. In other examples, the gap region and the features are discrete, e.g., as in the case of a plurality of grooves separated by corresponding gap regions. The separation provided by the gap region may be partial separation or complete separation. The gap region may have a surface material different from the surface material of the features defined in the surface. For example, the features of the array may have a polymer coating and a primer in an amount or concentration exceeding the amount or concentration present in the gap region. In some examples, the polymer coating and the primer may not be present in the gap region.

[0084] As used herein, "nitrile oxide" means "R a C≡N+O-" group, where R a As defined herein. Examples of the preparation of nitrile oxides include in situ generation from aldoximes by treatment with chloroamide-T or by the action of a base on an imidic chloride [RC(Cl)=NOH] or by reaction between hydroxylamine and an aldehyde.

[0085] As used herein, "nitrone" means group, wherein R1, R2 and R3 may be R as defined herein a and R b Any one of the groups.

[0086] As used herein, "nucleotide" includes a nitrogenous heterocyclic base, a sugar, and one or more phosphate groups. Nucleotide is the monomeric unit of a nucleic acid sequence. In RNA, the sugar is ribose, and in DNA, the sugar is deoxyribose, i.e., a sugar lacking the hydroxyl group present at the 2' position in the ribose. The nitrogenous heterocyclic base (i.e., nucleobase) can be a purine base or a pyrimidine base. Purine bases include adenine (A) and guanine (G) and their modified derivatives or analogs. Pyrimidine bases include cytosine (C), thymine (T) and uracil (U) and their modified derivatives or analogs. The C-1 atom of deoxyribose is bonded to the N-1 of pyrimidine or the N-9 of purine. Nucleic acid analogs can have any one of a modified phosphate backbone, sugar, or nucleobase. Examples of nucleic acid analogs include, for example, universal bases or phosphate-sugar backbone analogs, such as peptide nucleic acids (PNA).

[0087] As used herein, a "photoacid generator" (PAG) is a molecule that releases protons when exposed to radiation. The PAG typically undergoes proton photodissociation irreversibly.

[0088] As used herein, a "primer" is defined as a single-stranded nucleic acid sequence (e.g., single-stranded DNA). Some primers, which can be referred to as amplification primers, are used as a starting point for template amplification and cluster generation. Other primers, which can be referred to as sequencing primers, are used as a starting point for DNA synthesis. The 5' end of a primer can be modified to allow for a coupling reaction with the functional groups of the polymer coating. A primer can be any number of base lengths in length and can include a variety of non-natural nucleotides. In one example, a sequencing primer is a short strand ranging from 10 to 60 bases, or 20 to 40 bases.

[0089] As used herein, a "spacer layer" refers to a material that bonds two components together. In some examples, the spacer layer can be, or can be in contact with, a radiation absorbing material that facilitates the bonding. The spacer layer can be present in a bonding region, e.g., a region on a substrate that is to be bonded to another material, which can be, by way of example, a spacer layer, a cover, another substrate, or the like or a combination thereof (e.g., a spacer layer and a cover). The bonding formed at the bonding region can be a chemical bond (as described above) or a mechanical bond (e.g., using fasteners or the like).

[0090] A "thiol" functional group refers to -SH.

[0091] As used herein, the terms "tetrazine" and "tetrazinyl" refer to a six-membered heteroaryl group comprising four nitrogen atoms. The tetrazine can be optionally substituted.

[0092] As used herein, a "tetrazole" refers to a five-membered heterocyclic group comprising four nitrogen atoms. The tetrazole can be optionally substituted.

[0093] Resin composition

[0094] In some examples disclosed herein, a resin composition comprises an epoxy resin matrix; a first photoacid generator comprising an anion having a molecular weight of less than about 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight of greater than about 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cation of the first photoacid generator and the second photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition.

[0095] In other examples disclosed herein, a resin composition includes an epoxy resin matrix; a first photoacid generator comprising a first anion having a first molecular weight; and a second photoacid generator comprising a second anion having a second molecular weight that is at least 50 g / mol (and in some cases at least 90 g / mol) greater than the first molecular weight; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light used to cure the resin composition.

[0096] In any of the examples disclosed herein, it may be desirable for one of the two different anions (e.g., the smaller anion) to diffuse at a faster rate than the other of the two different anions (e.g., the larger anion). In one example, the larger anion has a volume at least four times (4x) larger than the smaller anion (assuming equal density).

[0097] The epoxy resin matrix includes at least one epoxy material. Any suitable epoxy monomer or cross-linkable epoxy copolymer can be used as the epoxy material. The epoxy material can be selected from the group consisting of:

[0098] i) epoxy-functionalized silsesquioxanes (described further below);

[0099] ii) Trimethylolpropane triglycidyl ether:

[0100]

[0101] iii) Tetrakis(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane:

[0102]

[0103] iv) Copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane:

[0104]

[0105] (wherein the ratio of m:n is in the range of 8:92 to 10:90);

[0106] v) 1,3-bis[2-(3,4-epoxycyclohexyl)ethyl]tetramethyldisiloxane:

[0107]

[0108] vi) 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane:

[0109]

[0110] vii) 3,4-epoxycyclohexylmethyl-3,4-epoxycyclohexanecarboxylate:

[0111]

[0112] viii) Bis((3,4-epoxycyclohexyl)methyl)adipate:

[0113]

[0114] ix) 4-vinyl-1-cyclohexene 1,2-epoxide:

[0115]

[0116] x) Vinylcyclohexene dioxide:

[0117]

[0118] xi) 4,5-Epoxytetrahydrophthalic acid diglycidyl ester:

[0119]

[0120] xii) 1,2-Epoxy-3-phenoxypropane:

[0121]

[0122] xiii) Glycidyl methacrylate:

[0123]

[0124] xiv) 1,2-Epoxyhexadecane:

[0125]

[0126] xv) Poly(ethylene glycol) diglycidyl ether:

[0127]

[0128] (where n is in the range of 1 to 100);

[0129] xvi) Pentaerythritol glycidyl ether:

[0130]

[0131] xvii) Cyclohexane-1,2-dicarboxylic acid diglycidyl ester:

[0132]

[0133] xviii) Diglycidyl tetrahydrophthalate:

[0134] as well as

[0135] xix) Combinations thereof. When a combination is used, it is understood that any two or more of the listed epoxy resin materials may be used together in the resin composition.

[0136] Epoxy-functionalized silsesquioxanes include a silsesquioxane core functionalized with epoxy groups.

[0137] As used herein, the term "silsesquioxane" refers to a hybrid intermediate (RSiO) between silicon dioxide (SiO2) and organosilicon (R2SiO2). 1.5 ) chemical composition. Example silsesquioxanes include polyhedral oligomeric silsesquioxanes, which are available under the trade name (Hybrid Plastics). Examples of polyhedral oligomeric silsesquioxanes can be as described in Kehagias et al., Microelectronic Engineering 86 (2009), pp. 776-778, which is incorporated by reference in its entirety. The composition is of the formula [RSiO 3 / 2 ] n The organosilicon compound wherein the R groups may be the same or different.

[0138] The resin composition disclosed herein may include one or more different cage or core silsesquioxane structures as monomer units. For example, the polyhedral structure may be a T8 structure (polyoctahedral cage or core structure), such as: and is represented by: The monomer unit typically has eight functional group arms R1 to R8.

[0139] The monomer unit may have a cage structure with 10 silicon atoms and 10 R groups, referred to as T 10 , such as: Alternatively, the monomer unit may have a cage structure with 12 silicon atoms and 12 R groups, referred to as T 12 , such as: Silsesquioxane-based materials may alternatively include T6, T 14 or T 16 Cage structure.

[0140] The average cage content can be adjusted during synthesis and / or controlled by purification methods, and the cage size distribution of the monomer units can be used in the examples disclosed herein. As an example, any one of the cage structures can be present in an amount ranging from about 30% to about 100% of the total silsesquioxane monomer units used. Thus, silsesquioxane-based materials can include a mixture of silsesquioxane configurations.

[0141] Silsesquioxane-based materials can be a mixture of cage structures and can include open and partially open cage structures. For example, any epoxy silsesquioxane material described herein can be a mixture of discrete silsesquioxane cages and non-discrete silsesquioxane structures and / or incompletely condensed discrete structures (such as polymers, ladders, etc.). Partially condensed materials will include epoxy R groups as described herein at some silicon vertices, but some silicon atoms will not be substituted with epoxy R groups, but instead may be substituted with OH groups. In some examples, the silsesquioxane material comprises a mixture of various forms, such as:

[0142] Condensation Cage

[0143] (a)

[0144] Incomplete condensation cage

[0145] (b)

[0146] Non-caged contents are large and poorly defined structures

[0147] and / or (c)

[0148] In the examples disclosed herein, R1 to R8 or R 10 or R 12 At least one of the silsesquioxanes comprises an epoxy group, and thus, the silsesquioxane is referred to as an epoxy silsesquioxane (e.g., an epoxy polyhedral oligomeric silsesquioxane). In some aspects, the epoxy silsesquioxane comprises epoxy end groups. An example of this type of silsesquioxane is a glycidyl group having the structure

[0149]

[0150] Another example of this type of silsesquioxane is an epoxycyclohexylethyl functionalized silsesquioxane having the structure

[0151]

[0152] One example of an epoxy resin matrix disclosed herein includes an epoxy-functionalized polyhedral oligomeric silsesquioxane, wherein the epoxy-functionalized polyhedral oligomeric silsesquioxane is selected from the group consisting of: glycidyl-functionalized polyhedral oligomeric silsesquioxane, epoxycyclohexylethyl-functionalized polyhedral oligomeric silsesquioxane, and combinations thereof. This example may include the epoxy silsesquioxane material alone or in combination with an additional epoxy material selected from the group consisting of: trimethylolpropane triglycidyl ether; tetra(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; a copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4-epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexylmethyl-3,4-epoxycyclohexane diglycidyl ether; pentaerythritol glycidyl ether; cyclohexane-1,2-dicarboxylate; diglycidyl tetrahydrophthalate; and combinations thereof.

[0153] In other silsesquioxane examples, a majority of the arms, such as eight, ten, or twelve arms or R groups, include epoxy groups. In other examples, R1 to R8 or R 10 or R 12 are the same, and therefore, R1 to R8 or R 10 or R 12 Each of R1 to R8 or R 10 or R 12 are different, and therefore, R1 to R8 or R 10 or R 12 At least one of R1 to R8 or R 10 or R 12 At least one of the epoxy groups is a non-epoxy functional group, which is selected from the group consisting of azide / azide, thiol, poly (ethylene glycol), norbornene and tetrazine, or other groups such as alkyl, aryl, alkoxy and haloalkyl groups. In some aspects, the non-epoxy functional group is selected to increase the surface energy of the resin. In these other examples, the ratio of epoxy groups to non-epoxy groups is in the range of 7:1 to 1:7, or 9:1 to 1:9, or 11:1 to 1:11.

[0154] In the examples disclosed herein, the epoxy silsesquioxane may also be a modified epoxy silsesquioxane that includes a controlled radical polymerization (CRP) agent and / or a functional group R1 to R8 or R 10 or R 12 One or more of the functional groups of interest are incorporated into the resin or core or cage structure.

[0155] Regardless of whether a single epoxy material or a combination of epoxy materials is used in the epoxy resin matrix, the total amount of epoxy resin matrix in the resin composition ranges from about 93 mass % to about 99 mass % of the total solids.

[0156] With any of the exemplary epoxy materials disclosed herein, it is understood that the epoxy groups allow the monomer units and / or copolymers to polymerize and / or crosslink into a crosslinked matrix, initially using a combination of ultraviolet (UV) light and acid (generated by a photoacid generator).

[0157] Examples of resin compositions also include combinations of photoacid generators. In some examples of such combinations, the first photoacid generator comprises a cation and a small anion, i.e., an anion having a molecular weight of less than about 250 g / mol; and the second photoacid generator comprises a cation and a large anion, e.g., an anion having a molecular weight greater than about 300 g / mol, such as, for example, an anion ranging from about 300 g / mol to about 1,000 g / mol. In other examples of such combinations, the first photoacid generator comprises a cation and a small anion having a first molecular weight; and the second photoacid generator comprises a cation and a large anion having a molecular weight at least 50 g / mol greater than the first molecular weight.

[0158] At least one of the cations of the photoacid generator in the resin composition should exhibit strong absorption of the incident light (e.g., ultraviolet (UV) light) that will be used to cure the resin composition. High-power UV light sources (such as medium-pressure mercury (MPM) lamps and UV light-emitting diodes (LEDs)) can emit UV energy in a narrow wavelength range centered on a specific wavelength, or be filtered to limit exposure to the UV energy. The light at the center wavelength is referred to herein as incident light. In some examples, the specific wavelength (incident light) is about 365 nm.

[0159] “Strongly absorbs,” “absorbs strongly,” and “absorbs strongly” mean that the cation of the first photoinitiator and / or the second photoinitiator has its maximum absorption (A) within + / - 40 nm of the wavelength of the incident light to be used to cure the resin composition. 最大 ), and / or the absorbance of the cation of the first photoinitiator and / or the second photoinitiator at the wavelength of the incident light to be used to cure the resin composition is greater than its maximum absorption (A 最大), and / or the cation of the first photoinitiator and / or the second photoinitiator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of the incident light used to cure the resin composition. In one example, the incident light has a wavelength in the range of about 350 nm to about 380 nm (with a center, for example, around 365 nm), and the cation of the first photoacid generator or the cation of the second photoacid generator has a mass attenuation coefficient in the range of about 0.9 L / (g*cm) to about 1.1 L / (g*cm) at the wavelength of the incident light.

[0160] Examples of cations that can strongly absorb incident light (such as light at about 365 nm) include triphenylsulfonium cations and cations with a maximum absorption wavelength (λ 最大 ) is a diaryliodonium cation of about 350 nm. The triphenylsulfonium cation may have the following structure:

[0161]

[0162] wherein R is a hydrogen atom and R' is:

[0163]

[0164] Maximum absorption wavelength (λ 最大 ) of about 350 nm may have the following structure:

[0165]

[0166] At least one of the desired cations exhibits strong absorption of incident light so that the photoacid generator in the resin composition is effectively activated when the resin is exposed to incident light. When one of the cations absorbs incident light, the excited cation enables its associated anion to produce an acid. The energy-absorbing cation can also transfer energy to the anion of the other photoacid generator, enabling the anion to produce an acid. When both cations absorb incident light, the corresponding cation enables its associated anion to produce an acid accordingly. When one or both cations effectively absorb light, the photoacid generator can produce the desired level of its corresponding acid.

[0167] In some examples of the resin composition, the first photoacid generator and the second photoacid generator both have the same cation. In these examples, both cations exhibit strong absorption of the incident light that will be used to cure the resin composition. The cation of the first photoacid generator and the cation of the second photoacid generator have a corresponding mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition. In these examples, the cation of the first photoacid generator and the cation of the second photoacid generator can be selected from the group consisting of: maximum absorption wavelength (λ最大 ) are diaryliodonium cations and triphenylsulfonium cations of about 350 nm.

[0168] However, it should be understood that two strongly absorbing cations may also increase the autofluorescence of the cured resin. Therefore, in cases where the first photoacid generator and the second photoacid generator have the same cation, it may be desirable to use a lower amount of each photoacid generator to avoid an increase in autofluorescence. In these examples, the first photoacid generator can be present in the resin composition in an amount ranging from about 1 mass % to about 1.3 mass % of the total solids in the resin composition, and the second photoacid generator can be present in the resin composition in an amount ranging from about 0.125 mass % to about 1.3 mass % of the total solids in the resin composition.

[0169] In other examples of the resin composition, the first photoacid generator and the second photoacid generator have different cations. One of the cations can strongly absorb incident light, while the other of the cations minimally absorbs or does not absorb incident light. The example of the cation that minimally absorbs or does not absorb incident light (such as light at about 365nm) includes bis-(4-methylphenyl) iodonium cation, (isopropylbenzene) cyclopentadienyl iron (II) cation, ferrocenium cation, 1-naphthyldiphenylsulfonium cation, (4-phenylthienyl) diphenylsulfonium cation, bis(2,4,6-trimethylphenyl) iodonium cation, bis(4-tert-butylphenyl) iodonium cation, N-hydroxy-5-norbornene-2,3-dicarboximide perfluoro-1-butanesulfonate cation and triarylsulfonium cation.

[0170] In some cases, it may be desirable for the second photoacid generator (including a large anion) to contain a cation that strongly absorbs incident light. As described in detail below, large anions tend to produce stronger acids, which can improve the kinetics of the polymerization reaction. Therefore, in some examples, the cation of the second photoacid generator (with a large anion) has a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition; and the cation of the first photoacid generator (with a small anion) has a mass attenuation coefficient of less than 0.1 L / (g*cm) at the wavelength of the incident light that cures the resin composition. As a specific example, the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大) is a diaryliodonium cation and a triphenylsulfonium cation with a wavelength of about 350 nm; and the cation of the first photoacid generator is selected from the group consisting of: a bis-(4-methylphenyl)iodonium cation, a (isopropylbenzene)cyclopentadienylferron (II) cation, a ferrocenium cation, a 1-naphthyldiphenylsulfonium cation, a (4-phenylthienyl)diphenylsulfonium cation, a bis(2,4,6-trimethylphenyl)iodonium cation, a bis(4-tert-butylphenyl)iodonium cation, an N-hydroxy-5-norbornene-2,3-dicarboximide perfluoro-1-butanesulfonate cation and a triarylsulfonium cation.

[0171] As briefly mentioned herein, when resin combination is exposed to incident light, strong absorbing cations absorb incident light, which effectively activates large anions and small anions to produce stronger acid and weaker acid respectively. Acid can extract electron pairs from the oxygen atom in the epoxy ring of epoxy material in the epoxy resin matrix. The oxygen atom can open the ring and make the polymer chain grow by attacking the more easily substituted carbon on the epoxy reactive group of the adjacent monomer and making the polymerization reaction grow.

[0172] Surprisingly, the combination of large and small anions, and their associated stronger and weaker acids, can synergistically influence the polymerization rate. Larger anions produce stronger acids, which are more reactive, and therefore increase the growth rate of each polymer chain. However, large anions may be diffusion-limited due to their size. In other words, large anions may limit the ability of living polymer chains to find more monomers to continue polymerization. Smaller anions produce weaker acids than large anions and therefore do not initially increase the propagation rate. However, smaller anions may be able to diffuse more efficiently through the partially cured resin, which has a higher viscosity and is more constrained than the uncured resin composition. Therefore, the smaller anion and its weaker acid can continue polymerization after the larger macroanion and its growing chain ends are captured. Therefore, small anions can promote higher monomer conversion rates than large anions. The combination of the initial rapid kinetics of large anions and the efficient diffusion of small anions appears to produce a surprising synergistic effect on the degree of cure without adversely affecting the hardness or autofluorescence properties of the cured resin.

[0173] The first photoacid generator comprises a small anion, which is defined herein as an anion having a molecular weight of less than about 250 g / mol. In some examples, the molecular weight of the small anion is in the range of about 140 g / mol to about 240 g / mol. Examples of suitable small anions include hexafluoroantimonate (SbF6 - ), hexafluorophosphate (PF6 - ) or hexafluoroarsenate (AsF6 -Hexafluoroantimonate has a molecular weight of approximately 235 g / mol. Hexafluorophosphate has a molecular weight of approximately 145 g / mol. Hexafluoroarsenate has a molecular weight of approximately 188 g / mol.

[0174] Some specific examples of the first photoacid generator include bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate (which can be used as PAG 270 is commercially available from IGM Resins), diaryliodonium hexafluorophosphate (available as SYLANTO TM 7MP is commercially available from Sylanto) and diaryliodonium hexafluoroantimonate (available as SYLANTO TM 7MS is commercially available from Sylanto).

[0175] The first photoacid generator may be included in the resin composition in an amount ranging from about 1% to about 5% by mass of the total solids in the resin composition. In another example, the first photoacid generator may be included in an amount ranging from about 2% to about 5% by mass of the total solids in the resin composition.

[0176] The second photoacid generator comprises a large anion, which in some examples is defined herein as an anion having a molecular weight greater than about 300 g / mol. In some examples, the molecular weight of the large anion is in the range of greater than about 300 g / mol to about 1,000 g / mol. In some examples, the molecular weight of the large anion is in the range of about 400 g / mol to about 900 g / mol, or about 600 g / mol to about 700 g / mol. Examples of suitable large anions include tetrakis(perfluorophenyl)borate ((C6F5)4B - ), tetrakis(pentafluorophenyl) gallate ((C6F5)4Ga - ) or tris[(trifluoromethyl)sulfonyl]methanide ((CF3SO2)2C - Tetrakis(pentafluorophenyl) gallate has a molecular weight of about 740 g / mol. Tris[(trifluoromethyl)sulfonyl]methanide has a molecular weight of about 411 g / mol.

[0177] Specific examples of the second photoacid generator include triphenylsulfonium tetrakis(perfluorophenyl)borate. Triphenylsulfonium tetrakis(perfluorophenyl)borate is available under the trade name PAG 290 (from BASF Corp.) is commercially available.

[0178] In other examples, the first and second photoacid generators are selected such that the molecular weight of one of the photoacid generators is at least 50 g / mol greater than the molecular weight of the other photoacid generator. In this example, any combination of anions can be used as long as the difference in molecular weight is at least 50 g / mol. Thus, in one example, the first photoacid generator may include a hexafluorophosphate anion (PF6 - ) (which has a molecular weight of about 145 g / mol), and the second photoacid generator may include a hexafluoroantimonate anion (SbF6 - ) (which has a molecular weight of about 235 g / mol). In this example, any combination of cations can be used as long as one cation has strong absorption of incident light.

[0179] In any of the examples disclosed herein, the second photoacid generator can be included in the resin composition in an amount ranging from about 0.1% to about 2% by mass of the total solids in the resin composition. In another example, the second photoacid generator can be included in an amount ranging from about 0.25% to about 1.5% by mass of the total solids in the resin composition.

[0180] The resin composition may also include surface additives. Surface additives can adjust the surface tension of the resin composition, which can improve the releasability of the resin from the imprinting device (e.g., a working stamp), improve the coatability of the resin composition, promote film stability, and / or improve leveling. Examples of surface additives include polyacrylate polymers (such as those available from BYK). ). The amount of the surface additive may be 3% by mass or less.

[0181] A specific example of a resin composition includes an epoxy resin matrix; a first photoacid generator comprising an anion having a molecular weight of less than about 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight greater than about 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light used to cure the resin composition.

[0182] Another specific example of a resin composition includes an epoxy resin matrix comprising an epoxy-functionalized polyhedral oligomeric silsesquioxane; a first photoacid generator comprising an anion having a molecular weight of less than about 250 g / mol; and a second photoacid generator comprising an anion having a molecular weight in the range of about 300 g / mol to about 1,000 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light used to cure the resin composition.

[0183] Any example of a resin composition can be prepared by mixing an epoxy resin matrix component with a first photoacid generator and a second photoacid generator. To deposit the resin composition, these components (epoxy resin matrix component and photoacid generator) can be diluted in a suitable solvent (to achieve the desired viscosity for the deposition technique used), such as propylene glycol monomethyl ether acetate (PGMEA), toluene, dimethyl sulfoxide (DMSO), tetrahydrofuran (THF), etc. In one example, the concentration of the epoxy resin matrix in the solvent is in the range of about 15 weight percent (wt%) to about 56 weight percent, and the concentration of the photoacid generator combination in the solvent is in the range of about 0.15 weight percent to about 4 weight percent. While not being bound by any particular theory, it is believed that the upper limit may be higher depending on the respective solubility of the epoxy resin matrix and the photoacid generator in the selected solvent. In one example, the solvent is PGMEA. In the resin composition and solvent solution or mixture, the total concentration of the resin composition (including the epoxy resin matrix and the photoacid generator) may range from about 15 wt % to about 60 wt %, and the amount of the solvent may range from about 40 wt % to about 85 wt %.

[0184] Flow cell and method

[0185] Any of the examples of resin compositions disclosed herein can be used to form a flow cell.

[0186] Figure 1An example of a method 100 for patterning a resin composition to form a surface of a flow cell is shown in FIG. 1. As shown, one example of the method 100 includes depositing a resin composition on a substrate, the resin composition including an epoxy resin matrix; a first photoacid generator including an anion having a molecular weight of less than about 250 g / mol; and a second photoacid generator including an anion having a molecular weight of greater than about 300 g / mol, wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first and second photoacid generators have a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition (reference number 102); nanoimprinting the deposited resin composition using a working stamp (reference number 104); and exposing the nanoimprinted deposited resin composition to incident light at an energy dose in a range of about 0.5 J to about 10 J for a time of 30 seconds or less to form a cured, patterned resin (reference number 106).

[0187] The resulting flow cell surface includes a substrate and a cured, patterned resin on the substrate, the cured, patterned resin including recesses separated by gap regions, and the cured, patterned resin having been formed from an example of the resin composition disclosed herein.

[0188] In some examples, the method 100 further includes functionalizing the recesses for a particular application, such as sequencing. In some examples, the method 100 further includes functionalizing the recesses for a particular application, such as sequencing. Figures 2A to 2C The method 100 is shown schematically in FIG. 1. Some examples of the method 100 further include functionalizing the recesses for a particular application, such as sequencing. In some examples, the method 100 further includes functionalizing the recesses for a particular application, such as sequencing. Figure 2D and Figure 2E Examples of functionalization of the recesses are shown in FIGS. 2-4.

[0189] Figure 2A A substrate 12 is shown, and Figure 2B An example of a resin composition 14 is shown deposited on the substrate 12.

[0190] Examples of suitable substrates 12 include: epoxysiloxane, glass, modified or functionalized glass, plastic (including acrylic, polystyrene and copolymers of styrene and other materials, polypropylene, polyethylene, polybutylene, polyurethane, polytetrafluoroethylene (such as Teflon® available from Chemours ), cyclic olefin / cyclic olefin polymer (COP) (such as Zeonex® available from Zeon ), polyimide, etc.), nylon (polyamide), ceramic / ceramic oxide, silica, fused silica or silica-based materials, aluminum silicate, silicon and modified silicon (e.g., boron-doped p+ silicon), silicon nitride (Si3N4), silicon oxide (SiO2), tantalum pentoxide (Ta2O5) or other tantalum oxides (TaO x), hafnium oxide (HfO2), carbon, metal, inorganic glass, etc. The substrate 12 may also be glass or silicon with a coating of tantalum oxide or another ceramic oxide at the surface.

[0191] Some examples of substrate 12 may have surface-bound epoxy silane attached thereto, which may react with other resin composition components to form resin composition 14 (and cured resin composition 14 ′) on substrate 12 .

[0192] In one example, substrate 12 can have a diameter in the range of about 2 mm to about 300 mm, or a rectangular sheet or panel having a maximum dimension of up to about 10 feet (about 3 meters). In one example, substrate 12 is a wafer having a diameter in the range of about 200 mm to about 300 mm. In another example, substrate 12 is a die having a width in the range of about 0.1 mm to about 10 mm. While example dimensions have been provided, it should be understood that substrates 12 having any suitable dimensions can be used. In another example, a panel can be used as a rectangular substrate 12, which has a larger surface area than a 300 mm circular wafer.

[0193] The resin composition 14 can be any example of the resin composition described herein, which includes a combination of photoacid generators. The resin composition 14 can be deposited on the substrate 12 using any suitable application technique that can be manual or automatic. As an example, the deposition of the resin composition 14 can be performed using vapor deposition techniques, coating techniques, grafting techniques, etc. Some specific examples include chemical vapor deposition (CVD), spraying (e.g., ultrasonic spraying), spin coating, thick coating or dip coating, doctor blade coating, whipping distribution, aerosol printing, screen printing, micro-contact printing, inkjet printing, etc. In one example, spin coating is used.

[0194] The deposited resin composition 14 is then patterned using any suitable patterning technique. Figure 2B In the example shown, nanoimprint lithography is used to pattern the resin composition 14. After depositing the resin composition 14, the resin composition may be soft-baked to remove excess solvent. When performed, the soft bake may be performed after depositing the resin and before positioning the working stamp 20 in the resin, and may be performed at a relatively low temperature in the range of about 50°C to about 150°C for a time period of greater than 0 seconds to about 3 minutes. In one example, the soft bake time is in the range of about 30 seconds to about 2.5 minutes.

[0195] like Figure 2BAs shown, a nanoimprint lithography mold or working stamp 20 is pressed against a layer of resin composition 14 to create indentations in the resin composition 14. The working stamp 20 comprises a template for the desired pattern to be transferred to the resin composition 14. In other words, the resin composition 14 is recessed or perforated by the protrusions of the working stamp 20. The resin composition 14 can then be cured with the working stamp 20 in place.

[0196] For the resin composition 14 disclosed herein, curing can be achieved by exposing the nanoimprinted deposited resin composition 14 to incident light for 30 seconds or less at an energy dose in a range of about 0.5 J to about 10 J. The incident light can be actinic radiation, such as ultraviolet (UV) radiation. In one example, the majority of the emitted UV radiation can have a wavelength of about 365 nm.

[0197] In the examples disclosed herein, energy exposure helps decompose the second photoacid generator (including absorbing cations and large anions) into a strong acid (super acid) that initiates polymerization and / or crosslinking of the epoxy resin matrix. Energy exposure also helps decompose the first photoacid generator into a weak acid that continues polymerization and / or crosslinking of the epoxy resin matrix through direct energy absorption by its cation or through indirect energy transfer from the cation of the second photoacid generator. With the effective degree of cure brought about by the photoacid generator combination described herein, the incident light exposure time can be 30 seconds or less. In some cases, the incident light exposure time can be 10 seconds or less. In other cases, the incident light exposure time can be about 3 seconds.

[0198] The curing process may include a single UV exposure stage. After curing and releasing the working stamp 20, topographical features, such as recessed portions 16, are defined in the resin composition 14. Figure 2C As shown, the resin composition 14 having recesses 16 defined therein is referred to as cured patterned resin 14'. Due at least in part to the efficient photopolymerization of the photoacid generator combinations disclosed herein, the methods disclosed herein do not involve a post-UV cure hard bake step to obtain a well-cured film.

[0199] The chemical composition of the cured patterning resin 14 ′ depends on the epoxy matrix and the photoacid generator used in the resin composition 14 .

[0200] like Figure 2C As shown, the cured patterned resin 14' includes recessed portions 16 defined therein and gap regions 22 separating adjacent recessed portions 16. In the examples disclosed herein, the recessed portions 16 are formed using a polymer hydrogel 18 ( Figure 2C and Figure 2D ) and primer 24 ( Figure 2E and Figure 2F) is functionalized, while portions of the interstitial regions 22 are available for bonding but will not have polymer hydrogel 18 or primer 24 thereon.

[0201] Many different layouts of the recesses 16 are contemplated, including regular, repeating, and irregular patterns. In one example, the recesses 16 are arranged in a hexagonal grid to achieve close packing and improved density. Other layouts may include, for example, linear (i.e., rectangular) layouts (e.g., troughs or grooves), triangular layouts, etc. In some examples, the layout or pattern may be an xy format of recesses 16 in rows and columns. In some other examples, the layout or pattern may be a repeating arrangement of recesses 16 and / or gap areas 22. In still other examples, the layout or pattern may be a random arrangement of recesses 16 and / or gap areas 22. The pattern may include stripes, swirls, lines, triangles, rectangles, circles, arcs, plaids, lattices, diagonal lines, arrows, squares, and / or cross hatching. In one example, as Figure 2C As shown, the recesses 16 are holes arranged in a row.

[0202] The layout or pattern of the recesses 16 can be characterized with respect to the density of the recesses 16 (ie, the number of recesses 16) in a defined area. For example, the recesses 16 can be approximately 2 million / mm 2 The density can be adjusted to different densities, including, for example, at least about 100 / mm 2 , about 1,000 pieces / mm 2 , about 100,000 / mm 2 , about 1 million / mm 2 , about 2 million pieces / mm 2 , about 5 million / mm 2 , about 10 million / mm 2 , about 50 million / mm 2 or greater or lesser density. It will also be understood that the density of the recesses 16 in the cured patterned resin 14' can be between a value selected from the lower limit and a value selected from the upper limit of the above range. By way of example, a high-density array can be characterized as having recesses 16 separated by less than about 100 nm, a medium-density array can be characterized as having recesses 16 separated by about 400 nm to about 1 μm, and a low-density array can be characterized as having recesses 16 separated by more than about 1 μm. While exemplary densities have been provided, it will be understood that a substrate having any suitable density can be used.

[0203] The layout or pattern of the recesses 16 can also be characterized based on or alternatively based on the average pitch from the center of the recess 16 to the center of the adjacent recess 16, i.e., the spacing (center-to-center spacing) or the average pitch from the right edge of one recess 16 to the left edge of the adjacent recess 16, i.e., the spacing (edge-to-edge spacing). The pattern can be regular, so that the coefficient of variation around the average pitch is small, or the pattern can be irregular, in which case the coefficient of variation can be relatively large. In either case, the average pitch can be, for example, at least about 10 nm, about 50 nm, about 0.1 μm, about 0.5 μm, about 1 μm, about 5 μm, about 10 μm, about 100 μm, or more or less. The average pitch of a particular pattern of recesses 16 can be between a value selected from the lower limit and a value selected from the upper limit of the above range. In one example, the recesses 16 have a pitch (center-to-center spacing) of about 1.5 μm. Although exemplary average pitch values ​​have been provided, it should be understood that other average pitch values ​​can be used.

[0204] The size of each recess 16 may be characterized by its volume, open area, depth, and / or diameter.

[0205] Each recess 16 can have any volume capable of confining a fluid. The minimum or maximum volume can be selected, for example, to accommodate the throughput (e.g., multiplexing), resolution, nucleotide or analyte reactivity desired for downstream use of the flow cell. For example, the volume can be at least about 1×10 -3 μm 3 , about 1×10 -2 μm 3 , about 0.1μm 3 , about 1μm 3 , about 10μm 3 , about 100μm 3 It should be understood that the polymer hydrogel 18 can fill all or part of the volume of the recess 16 .

[0206] The area occupied by each recess opening can be selected based on criteria similar to those set forth above for pore volume. For example, the area of ​​each recess opening can be at least about 1×10 -3 μm 2 , about 1×10 -2 μm 2 , about 0.1μm 2 , about 1μm 2 , about 10μm 2 , about 100μm 2 The area occupied by each recess opening may be greater than, less than, or in between the values ​​specified above.

[0207] The depth of each recess 16 can be large enough to accommodate a portion of the polymer hydrogel 18. In one example, the depth can be about 0.1 μm, about 0.5 μm, about 1 μm, about 10 μm, about 100 μm, or more or less. In some examples, the depth is about 0.4 μm. The depth of each recess 16 can be greater than, less than, or in between the values ​​specified above.

[0208] In some cases, the diameter or length and width of each recess 16 may be about 50 nm, about 0.1 μm, about 0.5 μm, about 1 μm, about 10 μm, about 100 μm, or more or less. The diameter or length and width of each recess 16 may be greater than, less than, or between the values ​​specified above.

[0209] like Figure 2C and Figure 2D As shown, after the resin composition 14 is patterned and cured, the cured patterned resin 14 ′ may be treated to prepare a surface for application of the polymer hydrogel 18 .

[0210] In one example, the cured patterned resin 14' can be exposed to a silanization reaction that attaches silane or a silane derivative to the cured patterned resin 14'. Silanization introduces the silane or silane derivative throughout the surface, including in the recesses 16 (e.g., on the bottom surface and along the sidewalls) and in the gap regions 22.

[0211] Silanization can be achieved using any silane or silane derivative. The choice of silane or silane derivative may depend in part on the polymer to be used to form the polymer hydrogel 18 (shown in FIG. Figure 2D The functionalized molecules of the present invention may be functionalized molecules (e.g., in the present invention) because it may be desirable to form a covalent bond between the silane or silane derivative and the polymer coating 18. The method for attaching the silane or silane derivative to the cured patterned resin 14' may vary depending on the silane or silane derivative used. Several examples are described herein.

[0212] Examples of suitable silanization methods include vapor deposition (eg, the YES method), spin coating, or other deposition methods. Some examples of methods and materials that can be used to silanize the cured patterned resin 14' are described herein, but it should be understood that other methods and materials can be used.

[0213] In an example utilizing a YES CVD oven, cured patterned resin 14' on substrate 12 is placed in the CVD oven. The chamber can be evacuated and the silanization cycle can begin. During the cycle, the silane or silane derivative container can be maintained at a suitable temperature (e.g., approximately 120°C for norbornene silane), the silane or silane derivative vapor line can be maintained at a suitable temperature (e.g., approximately 125°C for norbornene silane), and the vacuum line can be maintained at a suitable temperature (e.g., approximately 145°C).

[0214] In another example, silane or a silane derivative (e.g., liquid norbornene silane) can be deposited in a glass vial and placed in a glass vacuum desiccator with patterned substrate 12. The desiccator can then be evacuated to a pressure in the range of about 15 mTorr to about 30 mTorr and placed in an oven at a temperature in the range of about 60° C. to about 125° C. Silanization is allowed to proceed, and the desiccator is then removed from the oven, cooled, and vented to air.

[0215] Vapor deposition, the YES method, and / or a vacuum dryer can be used with various silanes or silane derivatives, such as those that include cycloolefin unsaturated moieties such as norbornene, norbornene derivatives (e.g., (hetero)norbornene including oxygen or nitrogen substituted for one of the carbon atoms), trans-cyclooctene, trans-cyclooctene derivatives, trans-cyclopentene, trans-cycloheptene, trans-cyclononene, bicyclo[3.3.1]non-1-ene, bicyclo[4.3.1]dec-1(9)-ene, bicyclo[4.2.1]non-1(8)-ene, and bicyclo[4.2.1]non-1-ene. Any of these cycloolefins can be substituted, for example, with R groups such as hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, or (heteroalicyclic)alkyl. Examples of norbornene derivatives include [(5-bicyclo[2.2.1]hept-2-enyl)ethyl]trimethoxysilane. As another example, these methods can be used when the silane or silane derivative includes a cycloalkyne unsaturated moiety, such as cyclooctyne, a cyclooctyne derivative, or a bicyclononyne (e.g., bicyclo[6.1.0]non-4-yne or a derivative thereof, bicyclo[6.1.0]non-2-yne, or bicyclo[6.1.0]non-3-yne). These cycloalkynes can be substituted with any of the R groups described herein.

[0216] The attachment of the silane or silane derivative forms a pre-treated (eg, silanized) cured patterned resin 14 ′ that includes silanized recessed portions and silanized interstitial regions.

[0217] In other examples, the cured patterning resin 14' may not be exposed to silanization. Instead, the cured patterning resin 14' may be exposed to plasma ashing, and then the polymer hydrogel 18 may be spin-coated (or otherwise deposited) directly onto the plasma-ashed cured patterning resin 14'. In this example, plasma ashing may produce surfactants (e.g., hydroxyl (C-OH or Si-OH) and / or carboxyl groups) that can adhere the polymer hydrogel 18 to the cured patterning resin 14'. In these examples, the polymer hydrogel 18 is selected so that it reacts with the surface groups produced by plasma ashing.

[0218] In yet other examples, the cured patterning resin 14' may include unreacted epoxy groups and, therefore, may not be exposed to silanization because the unreacted epoxy groups may react directly with the amino functional groups of the polymer hydrogel 18. In this example, for example, if it is desired to clean the surface of potential contaminants, plasma ashing may be performed.

[0219] The polymer hydrogel 18 can then be applied to the pre-treated cured patterned resin 14' (eg Figure 2C and Figure 2D ). The polymer hydrogel 18 may be a semi-rigid polymer material that is permeable to liquids and gases and tethered to the cured patterned resin 14'.

[0220] Examples of polymer hydrogels 18 include acrylamide copolymers, such as poly(N-(5-azidoacetamidopentyl)acrylamide-co-acrylamide), or PAZAM. Some other forms of PAZAM and acrylamide copolymers are represented by the following structure (I):

[0221]

[0222] in:

[0223] R A is selected from the group consisting of azide, optionally substituted amino, optionally substituted alkenyl, optionally substituted alkyne, halogen, optionally substituted hydrazone, optionally substituted hydrazine, carboxyl, hydroxy, optionally substituted tetrazole, optionally substituted tetrazine, nitrile oxide, nitrone, sulfate, and thiol;

[0224] R B is H or optionally substituted alkyl;

[0225] R C 、R D and R E are each independently selected from H and optionally substituted alkyl;

[0226] -(CH2) p- each of which may be optionally substituted;

[0227] p is an integer ranging from 1 to 50;

[0228] n is an integer ranging from 1 to 50,000; and

[0229] m is an integer within the range of 1 to 100,000.

[0230] It should be noted that the arrangement of recurring "n" and "m" features in structure (I) is representative, and that the monomer subunits may be present in any order in the polymer structure (e.g., random, block, patterned, or combinations thereof).

[0231] Other forms of acrylamide copolymers and PAZAM may have a molecular weight ranging from about 5 kDa to about 1500 kDa, or from about 10 kDa to about 1000 kDa, or in one specific example, about 312 kDa.

[0232] In some examples, the other forms of acrylamide copolymers and PAZAM are linear polymers. In some other examples, the other forms of acrylamide copolymers and PAZAM are lightly cross-linked polymers.

[0233] In other examples, the polymer hydrogel 18 can be a variation of structure (I). In one example, the acrylamide unit can be N,N-dimethylacrylamide. In this example, the acrylamide unit in structure (I) can be replaced by Replace, where R D 、R E and R F are each H or C1-C6 alkyl, and R G and R H Each is a C1-C6 alkyl group (rather than H, as in the case of acrylamide). In this example, q can be an integer in the range of 1 to 100,000. In another example, in addition to the acrylamide units, N,N-dimethylacrylamide can also be used. In this example, in addition to the recurring "n" and "m" features, structure (I) can also include where R D 、R E and R F are each H or C1-C6 alkyl, and R G and R H Each is a C1-C6 alkyl group. In this example, q can be an integer in the range of 1 to 100,000.

[0234] As another example of polymer hydrogel 18, the recurring "n" feature in structure (I) can be replaced with a monomer including a heterocyclic azido group having structure (II):

[0235]

[0236] where R 1 R2 is H or C1-C6 alkyl; L is a linker comprising a linear chain having 2 to 20 atoms selected from carbon, oxygen, and nitrogen, and 10 optional substituents on the carbon and any nitrogen atoms in the chain; E is a linear chain comprising 1 to 4 atoms selected from the group consisting of carbon, oxygen, and nitrogen, and optional substituents on the carbon and any nitrogen atoms in the chain; A is an N-substituted amide having H or C1-C4 alkyl attached to the N; and Z is a nitrogen-containing heterocycle. Examples of Z include 5 to 10 ring members present as a single cyclic structure or a fused structure. Some specific examples of Z include pyrrolidinyl, pyridinyl, or pyrimidinyl.

[0237] As yet another example, polymer hydrogel 18 may include recurring units of each of structures (III) and (IV):

[0238]

[0239] where R 1a 、R 2a 、R 1b and R 2b Each of R is independently selected from hydrogen, optionally substituted alkyl or optionally substituted phenyl; 3a and R 3b Each of L is independently selected from hydrogen, optionally substituted alkyl, optionally substituted phenyl, or optionally substituted C7-C14 aralkyl; and each L 1 and L 2 are independently selected from an optionally substituted alkylene linker or an optionally substituted heteroalkylene linker.

[0240] It should be understood that other molecules can be used to form the polymer hydrogel 18, as long as they are functionalized to graft the oligonucleotide primer 24 thereto. Other examples of suitable polymer layers include those with colloidal structures, such as agarose; or those with polymer network structures, such as gelatin; or those with cross-linked polymer structures, such as polyacrylamide polymers and copolymers, silane-free acrylamide (SFA) or azidated versions of SFA. Examples of suitable polyacrylamide polymers can be synthesized from acrylamide and acrylic acid or acrylic acid containing vinyl groups, or from monomers that form a [2+2] photocycloaddition reaction. Other examples of suitable polymer hydrogels 42 include mixed copolymers of acrylamide and acrylates. A variety of polymer architectures containing acrylic monomers (e.g., acrylamide, acrylates, etc.) can be used in the examples disclosed herein, such as branched polymers, including star polymers, star or star block polymers, dendrimers, etc. For example, monomers (e.g., acrylamide, acrylamide containing a catalyst, etc.) can be randomly or block-incorporated into the branches (arms) of the star-shaped polymer.

[0241] The polymer hydrogel 18 can be deposited on the surface of the pre-treated, cured patterned resin 14' using spin coating, dipping, dip coating, flow of functionalized molecules under positive or negative pressure, or another suitable technique. The polymer hydrogel 18 can be present in a mixture. In one example, the mixture includes water containing PAZAM or a mixture of ethanol and water containing PAZAM.

[0242] After coating, the polymer hydrogel 18 may also be exposed to a curing process to form a coating of the polymer hydrogel 18 over the entire patterned substrate (i.e., in the recessed portions 16 and on the gap regions 22). In one example, curing the polymer hydrogel 18 may be performed at a temperature in the range of room temperature (e.g., about 25° C.) to about 95° C. for a time in the range of about 1 millisecond to about several days. In another example, the time may be in the range of 10 seconds to at least 24 hours. In yet another example, the time may be in the range of about 5 minutes to about 2 hours.

[0243] The attachment of the polymer hydrogel 18 to the pre-treated recesses and interstitial regions can be performed by covalent bonding. The covalent attachment of the polymer hydrogel 18 to the silanized or plasma ashed recesses helps maintain the polymer hydrogel 18 in the recesses 16 throughout the life of the resulting flow cell during various uses. The following are some examples of reactions that can occur between silanes or silane derivatives and the polymer coating 18.

[0244] When the silane or silane derivative includes norbornene or a norbornene derivative as the unsaturated moiety, the norbornene or norbornene derivative can: i) undergo a 1,3-dipolar cycloaddition reaction with the azide / azido group of PAZAM; ii) undergo a coupling reaction with a tetrazine group attached to PAZAM; undergo a cycloaddition reaction with a hydrazone group attached to PAZAM; undergo a photoclick reaction with a tetrazole group attached to PAZAM; or undergo a cycloaddition reaction with a nitrile oxide group attached to PAZAM.

[0245] When the silane or silane derivative includes a cyclooctyne or a cyclooctyne derivative as the unsaturated moiety, the cyclooctyne or cyclooctyne derivative can: i) undergo a strain-promoted azide-alkyne 1,3-cycloaddition (SPAAC) reaction with the azide / azide group of PAZAM, or ii) undergo a strain-promoted alkyne-nitrile oxide cycloaddition reaction with a nitrile oxide group attached to PAZAM.

[0246] When the silane or silane derivative includes a bicyclononyne as the unsaturated moiety, the bicyclononyne can undergo similar SPAAC alkyne cycloaddition reactions with azides or nitrile oxides attached to PAZAM due to the strain in the bicyclic ring system.

[0247] To form the polymer hydrogel 18 within the recesses 16 and not within the interstitial regions 22 of the cured patterned resin 14', the interstitial regions 22 may be polished to remove the polymer hydrogel 18. The polishing process may be performed using a chemical slurry (including, for example, an abrasive, a buffer, a chelating agent, a surfactant, and / or a dispersant), which removes the polymer hydrogel 18 from the interstitial regions 22 without adversely affecting the underlying cured patterned resin 14' and / or substrate 12 at those regions. Alternatively, the polishing may be performed using a solution that does not include abrasive particles. The chemical slurry may be used in a chemical mechanical polishing system. In this example, a polishing head / pad or other polishing tool is capable of polishing the polymer hydrogel 18 from the interstitial regions 22 while leaving the polymer hydrogel 18 within the recesses 16 and leaving the underlying cured patterned resin 14' at least substantially intact. As an example, the polishing head may be a Strasbaugh ViPRR II polishing head. In another example, the polishing may be performed using a polishing pad and a solution without any abrasive. For example, the polishing pad can be used with a solution that does not contain abrasive particles (eg, a solution that does not include abrasive particles).

[0248] Figure 2DThe flow cell precursor 10 is shown after the polymer hydrogel 18 has been applied to the recess 16. The flow cell precursor 10 can be exposed to a cleaning process. This process can utilize a water bath and ultrasonic treatment. The water bath can be maintained at a relatively low temperature in the range of about 22°C to about 30°C. The silanized, coated, and polished patterned substrate can also be spin-dried or dried via another suitable technique.

[0249] like Figure 2D and Figure 2E As shown, the grafting process is performed so that the primer 24 (e.g., Figure 2F Two different primers 24, 24') are shown grafted to the polymer hydrogel 18 in the recess 16. The primers 24, 24' can be any forward amplification primer and / or reverse amplification primer. In this example, the primers 24, 24' are two different primers.

[0250] It is desirable that primers 24, 24' be immobilized to polymer hydrogel 18. In some examples, immobilization can be performed by single-point covalent attachment at the 5' end of the respective primers 24, 24' to polymer hydrogel 18. Any suitable covalent attachment means known in the art can be used. In some examples, immobilization can be performed by strong non-covalent attachment.

[0251] Examples of blocked primers that can be used include alkyne-terminated primers, tetrazine-terminated primers, azide-terminated primers, amino-terminated primers, epoxy- or glycidyl-terminated primers, phosphorothioate-terminated primers, thiol-terminated primers, aldehyde-terminated primers, hydrazine-terminated primers, phosphoramidite-terminated primers, triazolinedione-terminated primers, and biotin-terminated primers. In some specific examples, a succinimide (NHS) ester-terminated primer may react with an amine at the surface of the polymer hydrogel 18, an aldehyde-terminated primer may react with a hydrazine at the surface of the polymer hydrogel 18, or an alkyne-terminated primer may react with an azide at the surface of the polymer hydrogel 18, or an azide-terminated primer may react with an alkyne or DBCO (dibenzocyclooctyne) at the surface of the polymer hydrogel 18, or an amino-terminated primer may react with an activated carboxylic acid group or NHS ester at the surface of the polymer hydrogel 18, or a thiol-terminated primer may react with an alkylating reactant (e.g., iodoacetamide or maleimide) at the surface of the polymer hydrogel 18, a phosphoramidite-terminated primer may react with a thioether at the surface of the polymer hydrogel 18, or a primer modified with biotin may react with streptavidin at the surface of the polymer hydrogel 18.

[0252] Each of the primers 24, 24' has a universal sequence for capture and / or amplification purposes. Examples of primers 24, 24' include P5 and P7 primers, examples of which are used on the surface of commercial flow cells sold by Illumina Inc., for example, in HISEQ TM HISEQX TM 、MISEQ TM 、MISEQDX TM MINISEQ TM 、NEXTSEQ TM 、NEXTSEQDX TM 、NOVASEQ TM 、ISEQ TM 、GENOME ANALYZER TM and other instrument platforms.

[0253] For sequential paired end sequencing, each of these primers 24, 24 ' may also include a cleavage site. The cleavage sites of primers 24, 24 ' may be different from each other so that the cleavage of primers 24, 24 ' does not occur simultaneously. The example of a suitable cleavage site includes a nuclear base that can be enzymatically cleaved or a nuclear base that can be chemically cleaved, a modified nuclear base or a linker (e.g., between nuclear bases). The nuclear base that can be enzymatically cleaved may be easily cleaved by reacting with a glycosylase and an endonuclease, or an exonuclease. A specific example of a cleavable nuclear base is deoxyuracil (dU) that can be targeted by the USER enzyme. In one example, a uracil base can be incorporated into the 7th base position in the 3 ' end of a P5 primer (P5U) or a P7 primer (P7U). Other abasic sites may also be used. Examples of chemically cleavable nucleobases, modified nucleobases, or linkers include 8-oxoguanine, vicinal diols, disulfides, silanes, azobenzenes, photocleavable groups, allyl T (a thymidine nucleotide analog with allyl functionality), allyl ethers, or azide-functional ethers.

[0254] In one example, grafting can be achieved by flow-through deposition (e.g., using a temporarily bonded cap), thick coating, spraying, whipping dispensing, or another suitable method of attaching the primer 24, 24' to the polymer hydrogel 18. Each of these example techniques can utilize a primer solution or mixture that can include the primer 24, 24', water, a buffer, and a catalyst.

[0255] Thick coating may involve placing the flow cell precursor 10 (shown in Figure 2DIn one embodiment, the flow cell precursor 10 is immersed in a series of temperature-controlled baths. The baths may also be flow-controlled and / or covered with a nitrogen layer. The baths may include a primer solution or a mixture. In various baths, primers 24, 24' will be attached to primer-grafted functional groups of the polymer hydrogel 18 in at least some of the recesses 16. In one example, the flow cell precursor 10 is introduced into a first bath comprising a primer solution or a mixture, where a reaction occurs to attach primers 24, 24', and the flow cell precursor is then moved to an additional bath for washing. Moving between the baths may involve a robot arm or may be performed manually. A drying system may also be used in thick coatings.

[0256] Spray coating can be achieved by spraying the primer solution or mixture directly onto the flow cell precursor 10. The sprayed wafer can be incubated for a time in the range of about 4 minutes to about 60 minutes at a temperature in the range of about 0° C. to about 70° C. After incubation, the primer solution or mixture can be diluted and removed using, for example, a spin coater.

[0257] The whipping distribution can be performed according to the pooling and rotation separation method, and can therefore be achieved using a spin coater. The primer solution or mixture can be applied to the circulation cell precursor 10 (manually or via an automated process). The applied primer solution or mixture can be applied to the entire surface of the circulation cell precursor 10 or spread across the entire surface. The primer-coated circulation cell precursor 10 can be incubated for a time in the range of about 2 minutes to about 60 minutes at a temperature in the range of about 0°C to about 80°C. After incubation, the primer solution or mixture can be diluted and removed using, for example, a spin coater.

[0258] In other examples, the primers 24, 24' may be pre-grafted to the polymer hydrogel 18 and thus may be present in the recess 16 once the polymer hydrogel 18 is applied.

[0259] Figure 2E and Figure 2F An example of a flow cell 10 ′ after primer grafting is shown.

[0260] Figure 2E and Figure 2F The example shown is an example of a flow cell 10' without a cover or other flow cell 10' bonded thereto. In one example, the cover can be bonded to at least a portion of the cured patterned resin 14', for example, at some of the gap regions 22. The bond formed between the cover and the cured patterned resin 14' can be a chemical bond or a mechanical bond (e.g., using fasteners, etc.).

[0261] The cover can be any material that is transparent to the excitation light directed toward the substrate 12 and the cured patterned resin 14'. For example, the cover can be glass (e.g., borosilicate, fused silica, etc.), plastic, etc. A commercially available example of a suitable borosilicate glass is D Commercially available examples of suitable plastic materials (ie, cycloolefin polymers) are available from Zeon Chemicals LP. product.

[0262] The lid can be bonded to the cured patterned resin 14' using any suitable technique, such as laser bonding, diffusion bonding, anodic bonding, eutectic bonding, plasma activated bonding, glass frit bonding, or other methods known in the art. In one example, a spacer layer can be used to bond the lid to the cured patterned resin 14'. The spacer layer can be any material that seals at least some of the cured patterned resin 14' and the lid together. In some examples, the spacer layer can be a radiation absorbing material that facilitates bonding between the cured patterned resin 14' and the lid.

[0263] In other examples, two of the flow cells 10' can be bonded together such that the recess 16 faces the flow channel formed therebetween. The flow cells 10' can be bonded at the gap region 22 using similar techniques and materials as described herein for bonding the lids.

[0264] Methods of using a flow cell

[0265] The flow cell 10' disclosed herein can be used in a variety of sequencing methods or techniques, including techniques generally referred to as sequencing by synthesis (SBS), cycle array sequencing, sequencing by ligation, pyrosequencing, etc. In any of these techniques, since the polymer hydrogel 18 and attached primers 24, 24' are present in the recess 16 rather than in the interstitial region 22, amplification will be confined to the recess.

[0266] As an example, a sequencing-by-synthesis (SBS) reaction can be performed on a HISEQ instrument such as that from Illumina (San Diego, CA). TM HISEQX TM 、MISEQ TM 、MISEQDX TM MINISEQ TM 、NOVASEQ TM 、ISEQ TM 、NEXTSEQDX TM or NEXTSEQ TMA system on which the sequencer system operates. In SBS, the extension of nucleic acid primers (e.g., sequencing primers) along a nucleic acid template (i.e., sequencing template) is monitored to determine the sequence of nucleotides in the template. The underlying chemical process can be polymerization (e.g., catalyzed by a polymerase) or ligation (e.g., catalyzed by a ligase). In a particular polymerase-based SBS process, fluorescently labeled nucleotides are added to the sequencing primers in a template-dependent manner (thereby extending the sequencing primers) such that detection of the order and type of nucleotides added to the sequencing primers can be used to determine the sequence of the template.

[0267] Prior to sequencing, the capture and amplification primers 24, 24' can be exposed to a sequencing library that is amplified using any suitable method such as cluster generation.

[0268] In one example of cluster generation, the library fragments are copied from the hybridized primers 24, 24' by 3' extension using a high-fidelity DNA polymerase. The original library fragments are denatured, thereby immobilizing the copies. An isothermal bridge amplification can be used to amplify the immobilized copies. For example, the template loops of the copies hybridize to adjacent complementary primers 24, 24' and a polymerase copies the template to form a double-stranded bridge, the double-stranded bridge is denatured to form two single strands. The two strands loop back and hybridize to adjacent complementary primers 24, 24' and are again extended to form two new double-stranded loops. This process is repeated for each template copy through cycles of isothermal denaturation and amplification to produce a dense clonal cluster. Each cluster of double-stranded bridges is denatured. In one example, the antisense strands are removed by specific base cleavage, leaving the forward template polynucleotide strands. The clustering results in several template polynucleotide strands being formed in each of the recesses 16. This example of clustering is bridge amplification, and is one example of amplification that can be performed. It should be appreciated that other amplification techniques can be used, such as the Exclusion Amplification (ExAmp) workflow (Illumina Inc.).

[0269] A sequencing primer that hybridizes to a complementary sequence on the template polynucleotide strands can be introduced. This sequencing primer prepares the template polynucleotide strands for sequencing. The 3' ends of the primers 24, 24' (not attached to the copies) of the template and any flow cell binding can be blocked to prevent interference with the sequencing reaction, and in particular to prevent undesired priming.

[0270] To initiate sequencing, an incorporation mix can be added to the flow cell 10'. In one example, the incorporation mix includes a liquid carrier, a polymerase, and fluorescently labeled nucleotides. The fluorescently labeled nucleotides can include a 3' OH blocking group. When the incorporation mix is introduced to the flow cell 10', the fluid enters the flow channels and flows into the recesses 16 (where the template polynucleotide strands are present).

[0271] Fluorescently labeled nucleotides are added to the sequencing primer (thereby extending the sequence) in a template-dependent manner so that detection of the order and type of nucleotides added to the sequencing primer can be used to determine the sequence of the template. More specifically, one of the nucleotides is incorporated into a nascent chain that extends the sequencing primer and is complementary to the template polynucleotide chain by a corresponding polymerase. In other words, in at least some of the template polynucleotide chains across the entire flow cell 10', the corresponding polymerase extends the hybridized sequencing primer by incorporating one of the nucleotides in the mixture.

[0272] The incorporation of nucleotides can be detected by an imaging event.During an imaging event, an illumination system (not shown) can provide excitation light to the surface of the flow cell 10'.

[0273] In some examples, the nucleotide may further include a reversible termination property (e.g., a 3'OH blocking group) that terminates further primer extension once the nucleotide is added to the sequencing primer. For example, a nucleotide analog having a reversible terminator moiety may be added to the sequencing primer such that subsequent extension does not occur until a deblocking agent is delivered to remove the moiety. Thus, for examples using reversible termination, the deblocking agent may be delivered to the flow cell 10' after detection occurs.

[0274] Washing may occur between the various fluid delivery steps.The SBS cycle may then be repeated n times to extend the sequencing primer by n nucleotides, thereby detecting a sequence of length n.

[0275] In some examples, the forward strand can be sequenced and removed, and then the reverse strand constructed and sequenced as described herein.

[0276] While SBS has been described in detail, it will be appreciated that the flow cell 10 ′ described herein may be used for genotyping with other sequencing protocols, or in other chemical and / or biological applications.

[0277] Although Figure 1 and Figures 2A to 2F The examples described herein illustrate the use of an example resin composition to form a flow cell 10', but it should be understood that the resin compositions disclosed herein can be used in other applications requiring low autofluorescence. As an example, the resin compositions 14, 14' can be used in any optical-based SBS technology. As other examples, the resin compositions 14, 14' can be used in planar waveguides, in complementary metal oxide semiconductors (CMOS), and the like.

[0278] In order to further illustrate the present disclosure, examples are given herein. It should be understood that these examples are provided for illustrative purposes and should not be construed as limiting the scope of the present disclosure.

[0279] Non-limiting worked examples

[0280] Example 1

[0281] The comparative resin composition and the example resin composition of this example comprise an epoxy resin matrix of glycidyl polyhedral oligomeric silsesquioxane and epoxycyclohexyl polyhedral oligomeric silsesquioxane monomers.

[0282] Two of the comparative resin compositions (1 and 2) utilize a photoacid generator having a small anion (i.e., PAG 270 (triphenylsulfonium hexafluorophosphate)), and the other two of the comparative resin compositions (3 and 4) utilized a photoacid generator having a large anion (i.e., PAG 290 (triphenylsulfonium tetrakis(perfluorophenyl)borate) was prepared. An embodiment of the resin composition (5) was prepared using PAG 270 and Prepared in combination with PAG 290.

[0283] The resin compositions are provided in Table 1 below.

[0284] Table 1

[0285]

[0286] Each of the comparative resin and the example resin (e.g., approximately 18% by mass) was incorporated into a solvent mixture of PGMEA and DMSO. The resin / solvent mixture was spread and several manual indentations were made in each of the resins. The imprinted resin composition was cured (using 365nm UV light) using different exposure times, including 1 second, 3 seconds, 5 seconds, 7.5 seconds, 15 seconds, and / or 30 seconds.

[0287] The cured imprinted resin was analyzed using Fourier transform infrared spectroscopy (FTIR) and the results are shown in Figure 3 The Y axis represents the area at 2990 cm -1 The intensity at this wavelength is correlated with the hardness of the resin and, therefore, the degree of cure of the epoxy monomer. -1 A lower corrected intensity corresponds to a higher degree of cure. Figure 3 As shown, the sample of Example 5 is at 2990 cm -1 The corrected intensity at 2990 cm is less than that of each of Comparative Examples 1, 2, 3 and 4 at similar curing times. -1 The corrected intensity at 2990 cm-1 is about 0.11, while the corrected intensity at 2990 cm-1 for Comparative Examples 1 and 2 is about 0.11. -1The corrected intensities at 2990 cm and 300 cm were about 0.142 and 0.125, respectively. -1 The corrected intensities at 2990 cm-1 and 2990 cm-2 were about 0.146 and about 0.136, respectively. These results indicate that the Example resin composition (Example 5) having a combination of photoacid generators has the highest degree of cure (i.e., the highest degree of cure at 2990 cm-1) compared to the comparative resins having a photoacid generator with a small anion or a photoacid generator with a large anion. -1 The lowest correction strength is at the bottom) and the fastest curing degree.

[0288] Example 2

[0289] The comparative resin composition and the example resin composition of this example comprise an epoxy resin matrix of glycidyl polyhedral oligomeric silsesquioxane and epoxycyclohexyl polyhedral oligomeric silsesquioxane monomers.

[0290] Comparative resin (6) includes a photoinitiator (PI) (i.e., 2,2-dimethoxy-2-diphenylethanone) and a photoacid generator (i.e., bis-(4-methylphenyl)iodonium hexafluorophosphate). Example resin (7) includes the same epoxy resin matrix, but has an example of a photoacid combination disclosed herein instead of a PI / PAG combination. The photoacid combination in Example resin 7 includes a photoacid generator with a small anion (i.e., bis-(4-methylphenyl)iodonium hexafluorophosphate) and a photoacid generator with a large anion (i.e., PAG 290).

[0291] The resin compositions are provided in Table 2 below.

[0292] Table 2

[0293]

[0294] Each of the comparative resin and the example resin (e.g., about 18% by mass) was incorporated into a solvent mixture of PGMEA and DMSO. The tool imprints the resin / solvent mixture. The imprinted resin composition is cured using incident UV light at 365 nm at different doses (J). The dose for Example 7 ranges from 1 J to 10 J, while the dose for Comparative Example 6 ranges from 5 J to 60 J.

[0295] The cured imprinted resin was analyzed using FTIR and the results are shown in Figure 4 The Y axis represents the area at 2990 cm -1 The correction intensity at . Figure 4As shown, Example 7 had a UV radiation intensity of 2990 cm at each of the UV doses between 1 J (approximately 3.33 seconds depending on the tool used) and 10 J (approximately 33.3 seconds depending on the tool used). -1 The corrected intensity at 2990 cm is less than that of Comparative Example 6 at each of the UV doses between 5 J (about 16.65 seconds depending on the tool used) and 60 J (about 199.8 seconds depending on the tool used). -1 At all UV doses, Example 7 showed a positive correlation with the intensity at 2990 cm -1 The corrected intensity at 2990 cm-1 was about 0.10 or less, while the corrected intensity at 2990 cm-1 of Comparative Example 6 was about 0.10 or less. -1 The corrected intensities at 0.225 to about 0.12 are in the range of 0.225 to about 0.12. These results indicate that the example resin composition (Example 7) with the combination of photoacid generators achieves a higher degree of cure at a UV dose that is reduced by more than an order of magnitude when compared to the resin comprising the PI / PAG group.

[0296] A resin mixture including the resin of Example 7 was also deposited on 25 different glass wafers and imprinted using the master template. While the master template was held in place, the resin was cured using a 0.9 J dose (3 second cure time). The degree of cure was assessed using hardness measurements and FTIR for indentations 1, 5, 10, 15, and 20. The results are shown in Figure 5 These results show that the hardness values ​​(consistently between 0.23GPa and 0.25GPa) and the -1 The corrected IR intensities at φ are both stable for the different indentations (always between 0.090 and 0.100).

[0297] Quality measurements were also taken for recess depth, sidewall angle, top diameter, etc. The results were not reproduced herein, but were within expected values ​​for the master template used. The quality measurements showed that the example resins could pattern high-quality, fine features.

[0298] Example 3

[0299] The example resin composition of this embodiment includes an epoxy resin matrix of glycidyl polyhedral oligomeric silsesquioxane and epoxycyclohexyl polyhedral oligomeric silsesquioxane monomers.

[0300] The example resins (8A-8C, 9A-9C, and 10A-10C) comprised the same epoxy resin matrix and varying amounts of a photoacid generator having a small anion (i.e., bis-(4-methylphenyl)iodonium hexafluorophosphate) and a photoacid generator having a large anion (i.e., PAG 290). The resin compositions are provided in Table 3 below.

[0301] Table 3

[0302]

[0303] Each of the example resins (e.g., about 18% by mass) was incorporated into a solvent mixture of PGMEA and DMSO. The tool imprinted the resin / solvent mixture. The imprinted resin composition was cured (using incident UV light at 365 nm) using different exposure times including 2 seconds, 4 seconds, 8 seconds and 16 seconds.

[0304] Autofluorescence (AF) was measured for each of the cured imprinted resins. The results are shown in Figure 6 The Y axis represents the blue fluorescence intensity (in arbitrary units, au). Figure 6 As shown, regardless of the resin composition, the blue fluorescence intensity increases with the increase of the curing time. The blue intensity of Example 8A, Example 8B and Example 8C (each having 0.5% by mass of a photoacid generator with a large anion) is less than 10,000 at all curing times, indicating that a small amount of a photoacid generator with a large anion does not adversely affect the autofluorescence. When the mass % of the photoacid generator with a large anion is increased to 1 (Example 9A, Example 9B and Example 9C), regardless of the amount of the photoacid generator with a small anion, the blue color increases to above 10,000 at the longest curing time (16 seconds). This shows that the resin composition with a large amount of photoacid generator may perform better in terms of lower autofluorescence at shorter curing times. When the mass % of the photoacid generator with a large anion was increased to 1.5 (Examples 10A, 10B, and 10C), the blue color increased to above 10,000 at longer curing times (8 seconds and 16 seconds), regardless of the amount of the photoacid generator with a small anion. This also suggests that the resin composition with a large amount of photoacid generator may perform better in terms of lower autofluorescence at shorter curing times.

[0305] These cured imprinted resins were also analyzed using FTIR and the results are shown in Figure 7 The Y axis represents the area at 2990 cm -1 The correction intensity at . Figure 7 As shown, the photoacid generators of Example 8A, Example 8B, and Example 8C (each having 0.5 mass % of a photoacid generator having a large anion) at 2990 cm -1The corrected intensities at 2990 cm-1 are very similar at the corresponding cure times (e.g., about 0.125 at 2 seconds, about 0.10 at 4 seconds, between 0.08 and 0.085 at 8 seconds, and about 0.08 at 16 seconds). At both 2 and 4 second cure times, the presence of a large amount of PAG with a large anion appears to reduce the intensity at 2990 cm-1, regardless of the amount of PAG with a small anion. -1 More specifically, at 2 and 4 second cure times, each of Example 9A, Example 9B, Example 9C, Example 10A, Example 10B, and Example 10C exhibited a calibrated intensity at 2990 cm -1 The corrected intensity at 2990 cm was lower than that of each of Examples 8A, 8B, and 8C. However, at 8 and 16 second cure times, a large amount of a photoacid generator with a large anion was more effective when paired with a large amount of a photoacid generator with a small anion than when paired with a small amount of a photoacid generator with a small anion. More specifically, at 8 and 16 second cure times, each of Examples 9B, 9C, 10B, and 10C had a 0.1% peak at 2990 cm -1 The corrected intensity at is lower than that of Example 9A and Example 10A.

[0306] Figure 7 The peaks at 2990 cm-1 of each of the example resins -1 The corrected strength at the same curing time is also lower than Figure 3 These results illustrate the synergistic effects of the various embodiments of the photoacid generator combinations disclosed herein, compared to any of the photoacid generators used alone.

[0307] Additional Notes

[0308] It should be understood that all combinations of the foregoing concepts and the additional concepts discussed in more detail below (provided such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matter disclosed herein. Specifically, all combinations of the claimed subject matter appearing at the end of this disclosure are contemplated as being part of the inventive subject matter disclosed herein. It should also be understood that terminology explicitly employed herein that also appears in any disclosure incorporated by reference should be given a meaning most consistent with the specific concepts disclosed herein.

[0309] References throughout this specification to "one example," "another example," "an example," etc., mean that a particular element (e.g., feature, structure, and / or characteristic) described in conjunction with that example is included in at least one example described herein and may or may not be present in other examples. Furthermore, it should be understood that the elements described for any example may be combined in any suitable manner in the various examples, unless the context clearly indicates otherwise.

[0310] While several examples have been described in detail, it is to be understood that modifications may be made to the disclosed examples. Therefore, the above description should be considered non-limiting.

Claims

1. A resin composition, comprising: Epoxy resin matrix; a first photoacid generator selected from the group consisting of bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate; and a second photoacid generator comprising an anion having a molecular weight greater than 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; and wherein the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大 ) are diaryliodonium cations and triphenylsulfonium cations of 350 nm.

2. The resin composition of claim 1, wherein the epoxy resin matrix comprises an epoxy material selected from the group consisting of: epoxy-functionalized silsesquioxane; trimethylolpropane triglycidyl ether; tetrakis(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; a copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4-epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexyl methyl-3,4-epoxycyclohexanecarboxylate; bis((3,4-epoxycyclohexyl)methyl)adipate; 4-vinyl-1-cyclohexene-1,2-epoxide; vinylcyclohexene dioxide; diglycidyl-4,5-epoxytetrahydrophthalate; 1,2-epoxy-3-phenoxypropane; glycidyl methacrylate; 1,2-epoxyhexadecane; poly(ethylene glycol) diglycidyl ether; pentaerythritol glycidyl ether; diglycidyl cyclohexane-1,2-dicarboxylate; diglycidyl tetrahydrophthalate. 3 . The resin composition according to claim 1 , wherein the second photoacid generator is selected from the group consisting of triphenylsulfonium tetrakis(perfluorophenyl)borate, tetrakis(pentafluorophenyl)gallate, and tris[(trifluoromethyl)sulfonyl]methanate.

4. The resin composition according to claim 1, wherein: The cation of the second photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; and The cation of the first photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition.

5. The resin composition according to claim 1, wherein the cation of the first photoacid generator and the cation of the second photoacid generator have a corresponding mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition.

6. The resin composition according to claim 1, wherein: The first photoacid generator is present in an amount ranging from 1% to 5% by mass of the total solids in the resin composition; and The second photoacid generator is present in an amount ranging from 0.1% by mass to 2% by mass of the total solids in the resin composition. 7 . The resin composition according to claim 1 , wherein the molecular weight of the anion of the second photoacid generator is in a range of greater than 300 g / mol to 1,000 g / mol.

8. A resin composition, comprising: an epoxy resin matrix comprising an epoxy-functionalized polyhedral oligomeric silsesquioxane; a first photoacid generator selected from the group consisting of bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate; and a second photoacid generator comprising an anion having a molecular weight in a range of 300 g / mol to 1,000 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; and wherein the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大 ) are diaryliodonium cations and triphenylsulfonium cations of 350 nm. 9 . The resin composition according to claim 8 , wherein the epoxy-functionalized polyhedral oligomeric silsesquioxane is selected from the group consisting of glycidyl-functionalized polyhedral oligomeric silsesquioxane and epoxycyclohexylethyl-functionalized polyhedral oligomeric silsesquioxane.

10. The resin composition of claim 8, wherein the epoxy resin matrix further comprises an additional epoxy material selected from the group consisting of: trimethylolpropane triglycidyl ether; tetrakis(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; a copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4-epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexylmethyl- 3,4-Epoxycyclohexanecarboxylate; bis((3,4-epoxycyclohexyl)methyl)adipate; 4-vinyl-1-cyclohexene-1,2-epoxide; vinylcyclohexene dioxide; diglycidyl 4,5-epoxytetrahydrophthalate; 1,2-epoxy-3-phenoxypropane; glycidyl methacrylate; 1,2-epoxyhexadecane; poly(ethylene glycol) diglycidyl ether; pentaerythritol glycidyl ether; diglycidyl cyclohexane-1,2-dicarboxylate; diglycidyl tetrahydrophthalate. 11 . The resin composition according to claim 8 , wherein the first photoacid generator is present in an amount ranging from 1% by mass to 5% by mass of the total solids in the resin composition. 12 . The resin composition according to claim 8 , wherein the second photoacid generator is present in an amount ranging from 0.1% by mass to 2% by mass of the total solids in the resin composition. 13 . The resin composition according to claim 11 , wherein the second photoacid generator is selected from the group consisting of triphenylsulfonium tetrakis(perfluorophenyl)borate, tetrakis(pentafluorophenyl)gallate, and tris[(trifluoromethyl)sulfonyl]methanate.

14. The resin composition according to claim 8, wherein: The cation of the second photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; and The cation of the first photoacid generator has a mass attenuation coefficient of at least 0.1 L / (g*cm) at a wavelength of incident light that cures the resin composition.

15. The resin composition of claim 8, wherein the cation of the first photoacid generator and the cation of the second photoacid generator have a corresponding mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition.

16. A circulation cell, comprising: substrate; and a cured patterned resin on the substrate, the cured patterned resin comprising recessed portions separated by gap regions, and the cured patterned resin comprising a cured form of a resin composition comprising: Epoxy resin matrix; a first photoacid generator selected from the group consisting of bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate; and a second photoacid generator comprising an anion having a molecular weight greater than 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; and wherein the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大 ) are diaryliodonium cations and triphenylsulfonium cations of 350 nm.

17. The flow cell according to claim 16, further comprising: a hydrogel in the recess; as well as Amplification primers are attached to the hydrogel.

18. The flow cell of claim 16, wherein the epoxy resin matrix comprises an epoxy material selected from the group consisting of: epoxy-functionalized silsesquioxanes; trimethylolpropane triglycidyl ether; tetrakis(epoxycyclohexylethyl)tetramethylcyclotetrasiloxane; a copolymer of (epoxycyclohexylethyl)methylsiloxane and dimethylsiloxane; 1,3-bis[2-(3,4-epoxycyclohexyl)ethyl]tetramethyldisiloxane; 1,3-bis(glycidyloxypropyl)tetramethyldisiloxane; 3,4-epoxycyclohexyl methyl-3,4-epoxycyclohexanecarboxylate; bis((3,4-epoxycyclohexyl)methyl)adipate; 4-vinyl-1-cyclohexene-1,2-epoxide; vinylcyclohexene dioxide; diglycidyl-4,5-epoxytetrahydrophthalate; 1,2-epoxy-3-phenoxypropane; glycidyl methacrylate; 1,2-epoxyhexadecane; poly(ethylene glycol) diglycidyl ether; pentaerythritol glycidyl ether; diglycidyl cyclohexane-1,2-dicarboxylate; diglycidyl tetrahydrophthalate.

19. The flow cell of claim 16, wherein: The second photoacid generator is selected from the group consisting of triphenylsulfonium tetrakis(perfluorophenyl)borate, tetrakis(pentafluorophenyl)gallate, and tris[(trifluoromethyl)sulfonyl]methanide.

20. A method of manufacturing a flow cell, the method comprising: A resin composition is deposited on a substrate, the resin composition comprising: Epoxy resin matrix; a first photoacid generator selected from the group consisting of bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate; and a second photoacid generator comprising an anion having a molecular weight greater than 300 g / mol; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; wherein the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大 ) is a diaryliodonium cation and a triphenylsulfonium cation of 350 nm; nanoimprinting the deposited resin composition using a working stamp; and The nanoimprinted deposited resin composition is exposed to incident light at an energy dose ranging from 0.5 J to 10 J for 30 seconds or less to form a cured patterned resin.

21. A resin composition comprising: Epoxy resin matrix; a first photoacid generator selected from the group consisting of bis-(4-methylphenyl)iodonium hexafluorophosphate, triphenylsulfonium hexafluorophosphate, diaryliodonium hexafluorophosphate, and diaryliodonium hexafluoroantimonate; and a second photoacid generator comprising a second anion having a second molecular weight at least 50 g / mol greater than the molecular weight of the anion of the first photoacid generator; wherein i) the cation of the first photoacid generator, or ii) the cation of the second photoacid generator, or iii) the cations of the first photoacid generator and the second photoacid generator have a mass attenuation coefficient of at least 0.1 L / (g*cm) at the wavelength of incident light that cures the resin composition; and wherein the cation of the second photoacid generator is selected from the group consisting of: maximum absorption wavelength (λ 最大 ) are diaryliodonium cations and triphenylsulfonium cations of 350 nm.

Images