Two-dimensional photonic crystal material, functionalized photonic crystal hydrogel and preparation method thereof, and disease marker detection chip

Abstract

The application relates to the field of material and analytical chemistry, and provides a two-dimensional photonic crystal material, a functionalized photonic crystal hydrogel, a preparation method of the two-dimensional photonic crystal material and a disease marker detection chip. The preparation method of the two-dimensional photonic crystal material comprises the following steps: uniformly dispersing polymer colloidal microspheres in a first solvent to obtain a microsphere suspension; uniformly mixing the microsphere suspension with a second solvent to obtain a mixed solution; slowly injecting the mixed solution into a self-assembly container containing ultrapure water, so that the mixed solution is self-assembled on the liquid surface of the self-assembly container to form a two-dimensional photonic crystal array; transferring the self-assembly container to a closed gas chamber for organic solvent vapor annealing treatment; transferring the two-dimensional photonic crystal array on the liquid surface of the container to a glass slide subjected to hydrophilic treatment, and drying to obtain the two-dimensional photonic crystal material. The method can make the two-dimensional photonic crystal array more uniform and compact, so that the prepared material has higher lattice uniformity and better structural color saturation.

Description

Technical Field

[0001] This application relates to the fields of materials and analytical chemistry, and in particular to a two-dimensional photonic crystal material, a functionalized photonic crystal hydrogel and its preparation method, and a disease biomarker detection chip. Background Technology

[0002] Due to their excellent properties and potential applications in optics, electricity, heat, and magnetism, photonic crystals are increasingly favored by researchers as a novel material. Based on the periodicity of their spatial structure distribution and bandgap characteristics, photonic crystals can be classified into one-dimensional, two-dimensional, and three-dimensional photonic crystals.

[0003] Traditional two-dimensional photonic crystal materials have complex fabrication processes and are difficult to operate. Furthermore, the resulting materials have uneven distribution of photonic crystal arrays, low structural color saturation, and poor lattice uniformity.

[0004] Therefore, existing technologies still need improvement and development. Summary of the Invention

[0005] In view of this, in the first aspect, this application provides a two-dimensional photonic crystal material, a functionalized photonic crystal hydrogel and its preparation method, and a disease biomarker detection chip, aiming to solve the problems that the preparation process of traditional two-dimensional photonic crystal materials is relatively complex and difficult to operate, and the resulting materials have uneven distribution of photonic crystal arrays, low structural color saturation, and poor lattice uniformity.

[0006] To achieve the above-mentioned objectives, the technical solution adopted in this application is as follows: In a first aspect, embodiments of this application provide a method for preparing a two-dimensional photonic crystal material, including: The polymer colloidal microspheres were uniformly dispersed in the first solvent to obtain a microsphere suspension; The microsphere suspension was mixed evenly with the second solvent to obtain a mixed solution; The mixed solution is slowly injected into a self-assembly container filled with ultrapure water, so that the mixed solution self-assembles on the liquid surface of the self-assembly container to form a two-dimensional photonic crystal array. The self-assembled container was transferred to a sealed chamber for organic solvent vapor annealing. Then, the two-dimensional photonic crystal array on the liquid surface of the container was transferred to a hydrophilically treated glass slide and dried to obtain the two-dimensional photonic crystal material. In the organic solvent vapor annealing step, the organic solvent saturated vapor used is any one or more of toluene saturated vapor, xylene saturated vapor, tetrahydrofuran saturated vapor, dichloromethane saturated vapor, or acetone saturated vapor.

[0007] Secondly, embodiments of this application provide a two-dimensional photonic crystal material, which is prepared by the method for preparing two-dimensional photonic crystal materials in the first aspect.

[0008] Thirdly, embodiments of this application provide a method for preparing a functionalized photonic crystal hydrogel, comprising: Preparation of gel prepolymer solution; The gel prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light for photopolymerization to obtain a functionalized photonic crystal hydrogel. The functionalized photonic crystal hydrogel has specific recognition sites for detecting disease biomarkers. The forming container includes a first barrier layer and a second barrier layer that are arranged at relatively intervals; a photonic crystal interlayer is disposed between the first barrier layer and the second barrier layer; the photonic crystal interlayer includes a two-dimensional photonic crystal material in the second aspect.

[0009] Fourthly, embodiments of this application also provide a functionalized photonic crystal hydrogel, which is prepared by the method for preparing functionalized photonic crystal hydrogels in the third aspect.

[0010] Fifthly, embodiments of this application also provide a disease biomarker detection chip, the disease biomarker detection chip comprising: Accommodating cavity; The cavity contains at least one detection area, which includes a sample inlet and at least one detection chamber. The sample inlet is connected to each detection chamber via a microfluidic channel. The detection chambers are independent of each other and are not interconnected. Each detection chamber includes the functionalized photonic crystal hydrogel prepared in the fourth aspect.

[0011] The beneficial effects of this application include at least the following: The method for preparing two-dimensional photonic crystal materials provided in this application has a simple overall process and low operational difficulty. By transferring the self-assembly container to a closed gas chamber for organic solvent vapor annealing, the distribution of the two-dimensional photonic crystal array can be made more uniform and compact, resulting in a material with better structural color saturation and higher lattice uniformity. Attached Figure Description

[0012] To more clearly illustrate the technical solutions in the embodiments of this application, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0013] Figure 1These are physical images and schematic diagrams of the Debye diffraction testing apparatus provided in the embodiments of this application. Figure 2 This is a schematic diagram of the structure of a molded container provided in an embodiment of this application; Figure 3 This is a schematic diagram of the structure of a disease biomarker detection chip provided in one embodiment of this application; Figure 4 This is a scanning electron microscope image of the two-dimensional photonic crystal material prepared in Example 1 of this application; Figure 5 This is a schematic diagram of a two-dimensional photonic crystal array made of the two-dimensional photonic crystal material in Comparative Example 1; Figure 6 This is a schematic diagram of a two-dimensional photonic crystal array of the two-dimensional photonic crystal material prepared in Embodiment 1 of this application; Figure 7 This is a color diagram of the two-dimensional photonic crystal array structure of the two-dimensional photonic crystal material prepared in Example 1 of this application; Figure 8 This is a line graph showing the change of the Debye ring width of the two-dimensional photonic crystal array of the two-dimensional photonic crystal material prepared in Example 1 of this application as a function of annealing time; Figure 9 This is a top-view scanning electron microscope image of the functionalized photonic crystal hydrogel prepared in Example 2 of this application after freeze-drying; Figure 10 This is a cross-sectional scanning electron microscope image of the functionalized photonic crystal hydrogel prepared in Example 2 of this application; Figure 11 This is a scanning electron microscope image of the hydrogel network of the functionalized photonic crystal hydrogel prepared in Example 2 of this application; Figure 12 This is a physical image of the functionalized photonic crystal hydrogel prepared in Example 2 of this application; Figure 13 This is a photograph of the functionalized photonic crystal hydrogel prepared in Example 2 of this application after being cut into 7.5mm square pieces; Figure 14 This is a fitting curve of the lattice spacing change in response to different concentrations of glucose for the functionalized photonic crystal hydrogel prepared in Example 2 of this application; Figure 15 This is a fitting curve of the lattice spacing change in response to different concentrations of creatinine for the functionalized photonic crystal hydrogel prepared in Example 3 of this application; Figure 16 This is a graph showing the lattice spacing changes of functionalized photonic crystal hydrogels with different concentrations of urease prepared in Example 4 of this application before and after responding to urea. Figure 17 This is a fitting curve of the lattice spacing change in response to different concentrations of urea for the functionalized photonic crystal hydrogel prepared in Example 4 of this application. Figure 18 This is a graph showing the lattice spacing changes of functionalized photonic crystal hydrogels with different concentrations of anti-albumin antibodies prepared in Example 5 of this application before and after responding to albumin. Figure 19 This is a fitting curve of the lattice spacing change in response to different concentrations of albumin by the functionalized photonic crystal hydrogel prepared in Example 5 of this application; Figure 20 This is a schematic diagram of the detection process for simultaneously detecting four disease biomarkers using the detection chip provided in Embodiment 6 of this application. Detailed Implementation

[0014] To make the technical problems, technical solutions, and beneficial effects to be solved by this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are only for explaining this application, but the implementation of this application is not limited thereto.

[0015] Unless otherwise defined, the technical terms used in the following embodiments have the same meanings as commonly understood by those skilled in the art to which this application pertains. Unless otherwise specified, the experimental reagents used in the following embodiments are conventional biochemical reagents; the amounts of experimental reagents used are, unless otherwise specified, the amounts used in conventional experimental operations; and the experimental methods used are, unless otherwise specified, conventional methods.

[0016] Traditional processes for preparing two-dimensional photonic crystal materials are complex and difficult to operate. Furthermore, the resulting materials have uneven distribution of the two-dimensional photonic crystal array, low structural color saturation, and poor lattice uniformity.

[0017] In view of this, the present application provides a method for preparing two-dimensional photonic crystal materials. The method has a simple overall preparation process and low operation difficulty. By transferring the self-assembly container to a closed gas chamber for organic solvent vapor annealing, the distribution of the two-dimensional photonic crystal array can be made more uniform and compact, thereby the material obtained has better structural color saturation and higher lattice uniformity.

[0018] In a first aspect, embodiments of this application provide a method for preparing a two-dimensional photonic crystal material, including: The polymer colloidal microspheres were uniformly dispersed in the first solvent to obtain a microsphere suspension; The microsphere suspension was mixed evenly with the second solvent to obtain a mixed solution; The mixed solution is slowly injected into a self-assembly container filled with ultrapure water, so that the mixed solution self-assembles on the liquid surface of the self-assembly container to form a two-dimensional photonic crystal array. The self-assembled container was transferred to a sealed chamber for organic solvent vapor annealing. Then, the two-dimensional photonic crystal array on the liquid surface of the container was transferred to a hydrophilically treated glass slide and dried to obtain the two-dimensional photonic crystal material. In the organic solvent vapor annealing step, the organic solvent saturated vapor used is any one or more of toluene saturated vapor, xylene saturated vapor, tetrahydrofuran saturated vapor, dichloromethane saturated vapor, or acetone saturated vapor.

[0019] The method for preparing two-dimensional photonic crystal materials provided in this application is simple in process and easy to operate. By transferring the self-assembly container to a closed gas chamber for organic solvent vapor annealing, the distribution of the two-dimensional photonic crystal array can be made more uniform and compact, resulting in a material with better structural color saturation and higher lattice uniformity.

[0020] In a preferred embodiment, the organic solvent saturated vapor is toluene saturated vapor. Compared with using other organic solvent saturated vapors, the material obtained by annealing in a closed chamber filled with toluene saturated vapor exhibits the best structural color saturation and the highest lattice uniformity.

[0021] In some embodiments, the first solvent is ultrapure water; the second solvent is n-propanol; the polymer colloidal microspheres are at least one of polystyrene microspheres or polymethyl methacrylate microspheres; and the volume ratio of the microsphere suspension to the second solvent is 1:1.

[0022] In some embodiments, the particle size of the polymer colloidal microspheres is 580~650 nm.

[0023] In some embodiments, the mass concentration of the microsphere suspension is 20%.

[0024] In some implementations, the slide is placed in a plasma cleaner for radio frequency treatment for 30 seconds, which can improve the hydrophilicity of the slide.

[0025] In some implementations, a small amount of surfactant (such as sodium dodecyl sulfate) is added from one edge of the self-assembly container before the mixed solution is slowly injected into the self-assembly container containing ultrapure water. This helps to promote a more compact arrangement of the two-dimensional photonic crystal array.

[0026] Secondly, embodiments of this application also provide a two-dimensional photonic crystal material, which is prepared by the preparation method of the two-dimensional photonic crystal material in the first aspect.

[0027] The two-dimensional photonic crystal material prepared by the preparation method provided in the embodiments of this application has a uniform lattice, good structural color saturation, and high lattice uniformity.

[0028] Please see Figure 1 Two-dimensional photonic crystal arrays are structures with a close-packed hexagonal array. When monochromatic light of a specific wavelength is perpendicularly irradiated onto the two-dimensional photonic crystal array, the microspheres interfere with the incident light due to the ordered arrangement of the array structure and the same order of magnitude as the wavelength of the monochromatic light, resulting in strong forward diffraction in the conical direction. A bright Debye ring can be observed below the array, and the diameter of the ring satisfies the Bragg diffraction formula (1) with respect to the diffraction angle α between the particle spacing (particle spacing). (1); In equation (1), λ The wavelength of the diffracted light. d denoted as the lattice spacing of the photonic crystal.

[0029] The Bragg diffraction formula can be transformed using trigonometric functions as shown in equation (2): (2); In equation (2), α can be determined according to... Calculated; d λ is the lattice spacing of a photonic crystal. laser λ is the wavelength of the diffracted laser, D is the diameter of the Debye diffraction ring, and h is the vertical distance between the two-dimensional photonic crystal array and the Debye diffraction ring.

[0030] By measuring the diameter D of the Debye diffraction ring and substituting it into the above equations (1) and (2), the particle spacing in the two-dimensional photonic crystal array, i.e., the lattice spacing, can be calculated.

[0031] The change in the diameter of the Debye diffraction rings can directly reflect the change in the lattice spacing of a two-dimensional photonic crystal array, resulting in a more accurate response.

[0032] Thirdly, embodiments of this application also provide a method for preparing a functionalized photonic crystal hydrogel, comprising: Preparation of gel prepolymer solution; The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light for photopolymerization to obtain a functionalized photonic crystal hydrogel. The functionalized photonic crystal hydrogel has specific recognition sites for detecting disease biomarkers.

[0033] As an example, please refer to Figure 2The molding container provided in this application embodiment includes a first barrier layer 201 and a second barrier layer 202 disposed at relatively intervals; a photonic crystal interlayer 203 is disposed between the first barrier layer 201 and the second barrier layer 202, forming a "sandwich" structure. The photonic crystal interlayer 203 includes the two-dimensional photonic crystal material 2031 prepared above and a gap layer 2032. The first barrier layer 201 and the second barrier layer 202 can be glass slides.

[0034] In some embodiments, the gel prepolymer is filled into the photonic crystal interlayer of the molding container, and then placed in a 10 mW cm⁻¹ power source. 2 The functionalized photonic crystal hydrogel was obtained by irradiating it with a UV crosslinker at a wavelength of 365 nm for 10-20 minutes. The UV irradiation time can be flexibly adjusted according to the formulation of the gel prepolymer solution.

[0035] In some embodiments, the functionalized photonic crystal hydrogel has a first specific recognition site for glucose detection. The first specific recognition site may be a phenylboronic acid group grafted onto the photonic crystal hydrogel.

[0036] Preparation of the gel prepolymer includes: Acrylamide (AM) and N,N'-methylenebisacrylamide (MBA) were dissolved in ultrapure water to prepare the first solution; The second solution was prepared by dissolving 3-acrylamidophenylboronic acid (3-AAPBA) in a third solvent; The photoinitiator was dissolved in a fourth solvent to prepare a third solution; The first solution, the second solution, and the third solution are mixed to obtain a gel prepolymer solution.

[0037] In some embodiments, the third solvent is dimethyl sulfoxide (DMSO); the fourth solvent is dimethyl sulfoxide (DMSO); and the photoinitiator is 2,2-diethoxyacetophenone (DEAP).

[0038] By employing the above method, a first specific recognition site (phenylboronic acid group) can be introduced into the photonic crystal hydrogel. This first specific site can respond to different concentrations of glucose and cause changes in the lattice spacing of the photonic crystal array, thereby enabling qualitative and quantitative detection of glucose at different concentrations, with a detection limit (LoD) of approximately 0.4005 mmol / L. 1 .

[0039] In some embodiments, the functionalized photonic crystal hydrogel has a second specific recognition site for detecting creatinine. This second specific recognition site may be a creatinine molecular imprinted site introduced into the photonic crystal hydrogel.

[0040] Preparation of the gel prepolymer includes: Acrylic acid (AA), hydroxyethyl methacrylate (HEMA), N,N'-methylenebisacrylamide (MBA) and creatinine (CRE) were dissolved in ultrapure water, and sodium hydroxide solution was added and mixed evenly to obtain the fourth solution. The photoinitiator is dissolved in the fifth solvent to prepare the fifth solution; The fourth solution was refrigerated for 24 hours and then brought to room temperature to form hydrogen bonds and electrostatic complexes. The fifth solution was then added and mixed thoroughly to obtain the gel prepolymer solution. The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then subjected to photopolymerization under ultraviolet light irradiation to obtain a functionalized photonic crystal hydrogel, comprising: The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light to carry out a photopolymerization reaction to obtain the photonic crystal hydrogel. A solution of methanol, acetic acid, and sodium dodecyl sulfate was mixed to obtain an eluent. The photonic crystal hydrogel was eluted multiple times using an elution buffer to obtain a functionalized photonic crystal hydrogel.

[0041] In some embodiments, methanol and acetic acid are first mixed at a volume ratio of 9:1 to obtain a first mixed solution, and then the first mixed solution is mixed with sodium dodecyl sulfate solution (mass concentration of 1%) at a volume ratio of 1:1 to obtain an eluent.

[0042] In some embodiments, the photonic crystal hydrogel is immersed in the eluent prepared above and sonicated to elute creatinine template molecules. The eluent is rotated periodically until creatinine is no longer detectable in the eluent, with an elution time of approximately 6 hours. The concentration of creatinine in the eluent can be determined using a UV-Vis spectrophotometer.

[0043] Using the above method, a second specific recognition site (creatinine molecular imprinting site) can be introduced into the photonic crystal hydrogel. This second specific recognition site can respond to different concentrations of creatinine and cause changes in the lattice spacing of the photonic crystal array, thereby enabling qualitative and quantitative detection of creatinine at different concentrations. The limit of detection (LoD) is approximately 0.2692 mg / mL. 1 .

[0044] In some embodiments, the functionalized photonic crystal hydrogel has a third specific recognition site for detecting urea. This third specific recognition site may be an immobilized urease site introduced into the photonic crystal hydrogel.

[0045] Preparation of the gel prepolymer includes: Acrylamide (AM), N,N'-methylenebisacrylamide (MBA) and acrylic acid (AA) were dissolved in water, and then a photoinitiator was added and mixed evenly to obtain a gel prepolymer solution. The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then subjected to photopolymerization under ultraviolet light irradiation to obtain a functionalized photonic crystal hydrogel, comprising: The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light to carry out a photopolymerization reaction to obtain the photonic crystal hydrogel. An activation solution was prepared by dissolving 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in phosphate buffer (PBS solution). The photonic crystal hydrogel was placed in an activation solution for activation treatment to obtain the activated photonic crystal hydrogel; the activation time was 60 min. The activated photonic crystal hydrogel was incubated in a urease solution to obtain a functionalized photonic crystal hydrogel. The incubation temperature was 37°C and the incubation time was 24 hours.

[0046] In some embodiments, the molar ratio of 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) to N-hydroxysuccinimide (NHS) is 1:1. The molar concentration of the activation solution is 10 mM.

[0047] By employing the above method, a third specific recognition site (immobilized urease site) can be introduced into the photonic crystal hydrogel. This third specific recognition site can respond to different concentrations of urea and cause changes in the lattice spacing of the photonic crystal array, thereby enabling qualitative and quantitative detection of different concentrations of urea, with a detection limit of approximately 93.1 µmol / L. -1 .

[0048] In some embodiments, the functionalized photonic crystal hydrogel has a fourth specific recognition site for detecting albumin. This fourth specific recognition site may be an immobilized albumin antibody site introduced into the photonic crystal hydrogel.

[0049] Preparation of the gel prepolymer includes: Acrylamide (AM), N,N'-methylenebisacrylamide (MBA) and acrylic acid (AA) were dissolved in water, and then a photoinitiator was added and mixed evenly to obtain a gel prepolymer solution. The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then subjected to photopolymerization under ultraviolet light irradiation to obtain a functionalized photonic crystal hydrogel, comprising: The prepolymer solution is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light to carry out a photopolymerization reaction to obtain the photonic crystal hydrogel. An activation solution was prepared by dissolving 1-ethyl-(3-dimethylaminopropyl)carbodiimide (EDC) and N-hydroxysuccinimide (NHS) in phosphate buffer (PBS solution). The photonic crystal hydrogel was placed in an activation solution for activation treatment to obtain the activated photonic crystal hydrogel; the activation time was 60 min. The activated photonic crystal hydrogel was incubated in an anti-albumin antibody solution to obtain a functionalized photonic crystal hydrogel; the incubation temperature was 37℃ and the incubation time was 24 hours.

[0050] In this way, a fourth specific recognition site (immobilized albumin antibody site) can be introduced into the photonic crystal hydrogel. This fourth specific recognition site can respond to different concentrations of albumin and cause changes in the lattice spacing of the photonic crystal array, thereby realizing the qualitative and quantitative detection of albumin at different concentrations, with a detection limit of approximately 0.384 mg / mL.

[0051] Fourthly, embodiments of this application also provide a functionalized photonic crystal hydrogel, which is prepared by the method for preparing functionalized photonic crystal hydrogels in the third aspect.

[0052] The functionalized photonic crystal hydrogel provided in this application embodiment changes volume after recognizing target molecules (disease biomarkers, such as glucose, albumin, urea, creatinine, etc.) in the sample being tested. This further causes a change in the lattice spacing of the upper two-dimensional photonic crystal material layer. By testing the change in the diameter of the Debye diffraction ring, the concentration of the target molecules in the sample being tested can be calculated, thereby achieving sensitive label-free detection of target molecules in the analyte.

[0053] Traditional methods for quantitative detection of disease biomarkers, such as developing kits for specific disease-related biomarkers in the human body and detecting them using ELISA readers or high-sensitivity immunoassay systems, are complex, costly, and time-consuming. Another method involves modifying photonic crystal materials with fluorescent probes that specifically bind to the biomarker. The fluorescence signal excited by the binding of the probe molecules to the biomarker is amplified by the photonic crystal material, achieving specific and sensitive detection of the biomarker. However, this method requires complex fluorescent probe design, strict preparation conditions, and fluorescence lifetime limited by various environmental factors. The detection process also relies on large-scale equipment such as fluorescence spectrometers, resulting in high costs.

[0054] Secondly, traditional quantitative detection methods for disease biomarkers are essentially single-detection systems, making it difficult to simultaneously meet the specific detection requirements of multiple biomarker molecules. Furthermore, signal interference can easily occur when different biomarker detection systems are integrated, leading to decreased specificity and thus affecting the accuracy and reliability of the detection results.

[0055] Furthermore, simple qualitative test strip methods cannot provide reliable measurement results.

[0056] It is evident that traditional quantitative detection methods for disease biomarkers are cumbersome, costly, and time-consuming, resulting in poor accuracy and reliability of the results. In contrast, simple qualitative test strip methods cannot provide reliable measurement results.

[0057] In view of this, in a fifth aspect, embodiments of this application also provide a disease biomarker detection chip, the disease biomarker detection chip comprising: a receiving cavity; at least one detection area is disposed within the receiving cavity, each detection area including a sample inlet and at least one detection chamber; the sample inlet is connected to each detection chamber via a microfluidic channel; each detection chamber is independent of each other and not connected to each other; each detection area is independent of each other and not connected to each other; each detection chamber includes the functionalized photonic crystal hydrogel prepared in the fourth aspect.

[0058] By setting at least one detection region within the containment cavity, functionalized hydrogels for detecting different biological samples and the same / different specific detection sites can be integrated into different detection regions of the same detection chip. This enables simultaneous quantitative detection of multiple disease biomarkers from different biological samples, significantly improving detection efficiency. Furthermore, the different detection regions are independent and unconnected to each other, thus avoiding cross-interference between different detection regions and improving the accuracy and reliability of the detection results.

[0059] By setting up a sample inlet and at least one detection chamber within a detection area, functionalized hydrogels with the same or different specific detection sites for detecting the same biological sample can be integrated into different detection chambers within the same detection area of ​​the same detection chip. This enables simultaneous quantitative detection of multiple disease biomarkers in the same biological sample, significantly improving detection efficiency. Furthermore, the different detection chambers are independent and not interconnected, thus avoiding cross-interference between chambers and improving the accuracy and reliability of the detection results.

[0060] In practical applications, the number of detection areas in the containment cavity, the number of detection chambers in each detection area, and the functionalized photonic crystal hydrogel in the detection chambers can be adjusted according to the actual situation to meet the detection needs of various scenarios.

[0061] Figure 3 This is a schematic diagram of the structure of a disease biomarker detection chip provided in one embodiment of this application. Please refer to [link / reference]. Figure 3 The disease biomarker detection chip includes a receiving cavity 30; the receiving cavity 30 has at least one detection area (e.g., detection area 1 to detection area N, where N is an integer ≥ 2), each detection area includes a sample inlet 301 and at least one detection chamber 302; the sample inlet 301 is connected to each detection chamber 302 via a microfluidic channel 303; each detection chamber 302 is independent and not interconnected; each detection chamber 302 contains the functionalized photonic crystal hydrogel prepared above. The same or different functionalized photonic crystal hydrogels can be placed in each detection chamber.

[0062] In some implementations, a detection area is used to detect the same biological sample. For example, if the biological sample to be detected includes blood, urine, sweat, intercellular fluid, and tears, then five detection areas can be set in the housing 30 of the disease biomarker detection chip, labeled as detection areas 1 to 5. Detection area 1 is used to detect relevant disease biomarkers present in blood, detection area 2 is used to detect relevant disease biomarkers present in urine, detection area 3 is used to detect relevant disease biomarkers present in sweat, detection area 4 is used to detect relevant disease biomarkers present in intercellular fluid, and detection area 5 is used to detect relevant disease biomarkers present in tears.

[0063] As an example, please refer to Figure 3 Assuming it is necessary to simultaneously detect four combined biomarkers of diabetic nephropathy—glucose, albumin, urea, and creatinine—present in urine, a detection area (e.g., detection area 1) can be set up within the housing 30 of the disease biomarker detection chip. Detection area 1 includes four detection chambers (labeled A, B, C, and D, respectively). Detection chamber A contains a functionalized photonic crystal hydrogel with a first specific recognition site for glucose detection; detection chamber B contains a functionalized photonic crystal hydrogel with a second specific recognition site for creatinine detection; detection chamber C contains a functionalized photonic crystal hydrogel with a third specific recognition site for urea detection; and detection chamber D contains a functionalized photonic crystal hydrogel with a fourth specific recognition site for albumin. The diluted sample solution of the biological sample to be tested is injected through the injection port. The diluted sample solution flows into detection chambers A, B, C and D through the microfluidic channel. After the reaction is completed, the average value of the Debye diffraction ring diameter of the functionalized photonic crystal hydrogel in detection chambers A, B, C and D is measured to obtain the quantitative concentration of each marker (glucose, albumin, urea and creatinine).

[0064] Using the disease biomarker detection chip provided in this application embodiment, simultaneous quantitative detection of multiple disease biomarkers can be achieved, greatly improving detection efficiency.

[0065] The disease biomarker detection chip provided in this application has high detection sensitivity, is label-free, and requires no complex instruments. Detection can be completed using a portable optical device, making it particularly suitable for home health monitoring, postoperative dynamic monitoring, and exercise metabolism analysis scenarios. It is easier to detect metabolic abnormalities than traditional single-index detection methods.

[0066] This application has undergone multiple experiments, and some of the test results are presented here for reference to further describe the invention in detail. The following is a detailed description in conjunction with specific embodiments.

[0067] Example 1 This embodiment provides a two-dimensional photonic crystal material, the preparation method of which includes the following steps: Polystyrene microspheres with a particle size of 580 nm to 650 nm were uniformly dispersed in a first solvent (ultrapure water) to prepare a microsphere suspension with a mass concentration of 20%. The microsphere suspension was mixed with a second solvent (n-propanol) at a volume ratio of 1:1 and sonicated for 30 minutes until completely mixed to obtain a mixed solution. The mixed solution was drawn into a syringe (taking care to remove air bubbles), and the syringe was connected to a Teflon tube using a tip connector (or a lumbar puncture needle), with the other end connected to a pipette tip. The syringe was then fixed to a microinjection pump. An appropriate amount of ultrapure water was added to a clean self-assembly container (such as a glass jar, which must be thoroughly cleaned before each use; otherwise, residual surfactants will make it difficult for the solution to spread on the water surface). The pipette tip of the device was held vertically and in close contact with the liquid surface. The syringe pump was then turned on. In injection mode, the syringe specifications were set to 1 mL, 4.699 mm, injection volume to 1 mL, and injection speed to 30 µL / min. The start switch was pressed, and the syringe pump slowly ejected the mixed solution at the set speed, delivering it to the liquid surface of the glass cylinder through the syringe tip. As the mixed solution gradually spread, the central blank circle gradually shrank, stopping when the circle diameter reached approximately 1 cm, and the syringe tip was lifted. A small amount of surfactant (such as sodium dodecyl sulfate) was added to one edge of the glass cylinder to make the two-dimensional photonic crystal array more compact. The glass cylinder was then transferred to a sealed chamber filled with toluene saturated vapor and left to stand for 10 minutes to enhance the lattice uniformity of the two-dimensional photonic crystal array through toluene annealing. A 24 mm × 24 mm glass slide was placed in a plasma cleaner and radio frequency treated for 30 seconds to improve its hydrophilicity. Finally, the hydrophilically treated glass slide was lowered from the water surface of the glass cylinder to retrieve the two-dimensional photonic crystal array, maintaining a certain tilt angle during retrieval and allowing it to air dry naturally, thus obtaining the two-dimensional photonic crystal material.

[0068] The two-dimensional photonic crystal material prepared in Example 1 was scanned and detected using a scanning electron microscope. The detection results are as follows: Figure 4 As shown.

[0069] Comparative Example 1 The preparation method of the two-dimensional photonic crystal material in this comparative example is basically the same as that in Example 1, except that the step of transferring the glass cylinder to a sealed gas chamber filled with toluene saturated vapor and letting it stand for 10 minutes is omitted, that is, the two-dimensional photonic crystal array is not annealed using toluene saturated vapor.

[0070] The morphology of the two-dimensional photonic crystal materials prepared in Example 1 and Comparative Example 1 was examined and analyzed using scanning electron microscopy. The results are as follows: Figures 5-6 As shown. Among them, Figure 5 , Figure 6 The inset in the upper right corner is an image without Fast Fourier Transform. From Figure 5 and Figure 6 It can be seen that the organic solvent saturated vapor annealing process can make the two-dimensional photonic crystal array more uniform and compact.

[0071] Figure 7 This is a color diagram of the two-dimensional photonic crystal array structure of the two-dimensional photonic crystal material prepared in Example 1; Figure 8 This is a line graph showing the change in Debye ring width of the two-dimensional photonic crystal array prepared in Example 1 as a function of annealing time. The two-dimensional photonic crystal array structure color map is a visualization method that directly observes the macroscopic color representation and microscopic structural order of the two-dimensional photonic crystal array using optical microscopy / scanning electron microscopy (SEM) combined with optical imaging. Debye rings are ring-shaped diffraction patterns produced by a two-dimensional photonic crystal array in X-ray diffraction (XRD) or electron diffraction (ED) tests, and their ring width (half-width at half maximum, FWHM) is a core quantitative indicator for measuring the order of the crystal / array. This line graph mainly characterizes the regulatory effect of annealing time on the order and crystallinity of the two-dimensional photonic crystal array. From... Figure 7 and Figure 8 It can be seen that using organic solvent saturated vapor annealing process can help improve the lattice uniformity and structural color saturation of the material.

[0072] Example 2 This embodiment provides a functionalized photonic crystal hydrogel, the preparation method of which includes the following steps: (1) Preparation of the molded container: Five layers of tape are attached to the two short sides of the glass slide (first barrier layer) carrying the two-dimensional photonic crystal array obtained in Example 1 above. Another clean glass slide (second barrier layer) is used to sandwich the glass slide carrying the two-dimensional photonic crystal array (photonic crystal sandwich layer) to form a molded container with a "sandwich" structure.

[0073] (2) Preparation of gel prepolymer solution: Weigh 500 mg of acrylamide (AM) and 25 mg of N,N'-methylenebisacrylamide (MBA) and dissolve them in 2.4 mL of ultrapure water to prepare the first solution. Weigh 100 mg of 3-acrylamidophenylboronic acid (3-AAPBA) and dissolve it in 600 µL of dimethyl sulfoxide (DMSO) to prepare the second solution. Dissolve 2,2-diethoxyacetophenone (DEAP) in dimethyl sulfoxide (DMSO) to prepare a third solution with a volume concentration of 10 v / v%. Thoroughly stir and mix the first, second, and third solutions prepared above, and sonicate them to obtain the gel prepolymer solution. After purging with nitrogen to remove oxygen, store it in the dark and refrigerated.

[0074] (3) When using, use a 1 mL syringe to draw about 0.2 mL of the gel prepolymer solution prepared in step (2) above, and fill it into the photonic crystal interlayer of the molded container prepared in step (1) above. Note that air bubbles should not be introduced, and a small amount of gel prepolymer solution should be reserved at the edges. Then place the molded container in a UV crosslinker and irradiate it with UV light for 10 min to carry out the photopolymerization reaction, so as to obtain a functionalized photonic crystal hydrogel with a first specific recognition site for glucose detection. Soak the functionalized photonic crystal hydrogel in a centrifuge tube containing PBS buffer (phosphate buffer solution with pH=8.5) and place it on a shaker for later use.

[0075] The concentration of 10 mmol / L was prepared using PBS buffer at pH 8.5. 1 The glucose standard solution was serially diluted to a concentration of 1.0 mmol / L. 1 2.0 mmol L 1 4.0 mmol L 1. 6.0 mmol / L 1. 8.0 mmol / L 1 A diluted glucose solution.

[0076] The scanning electron microscope image of the functionalized photonic crystal hydrogel prepared in Example 2 above is shown below. Figures 9-13 As shown.

[0077] The functionalized photonic crystal hydrogel prepared in Example 2 above was immersed in PBS buffer for 1 hour, and then... Figure 1 The test setup shown was used to test the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different locations was taken; then the samples were sequentially transferred to a concentration of 1.0 mmol L... 12.0 mmol L 1 4.0 mmol L 1 6.0 mmol / L 1 8.0 mmol / L 1 The sample was soaked in a diluted glucose solution for 1 hour, and the diameter of the Debye diffraction ring was determined and recorded using the same method. The test results are as follows: Figure 14 As shown.

[0078] from Figure 14 It can be seen that within the tested concentration range (1~10 mmol L), 1 The interparticle spacing of the functionalized photonic crystal hydrogel increases with increasing glucose concentration, and the interparticle spacing of the functionalized photonic crystal hydrogel shows a good linear correlation with glucose concentration (R0). 2 = 0.9895). The limit of detection (LoD) was calculated to be 0.4005 mmol / L based on the ratio of the sample mean standard deviation (σ) to the slope of the calibration curve (S). 1 .

[0079] Example 3 This embodiment provides a functionalized photonic crystal hydrogel, the preparation method of which includes the following steps: (1) Preparation of the molded container: Five layers of tape are attached to the two short sides of the glass slide (first barrier layer) carrying the two-dimensional photonic crystal array obtained in Example 1 above. Another clean glass slide (second barrier layer) is used to sandwich the glass slide carrying the two-dimensional photonic crystal array (photonic crystal sandwich layer) to form a molded container with a "sandwich" structure.

[0080] (2) Preparation of gel prepolymer: Under light-protected conditions, 2.08 mmol acrylic acid (AA), 0.42 mmol hydroxyethyl methacrylate (HEMA), 0.32 mmol N,N'-methylenebisacrylamide (MBA), and 0.5 mmol creatinine (CRE) were dissolved in ultrapure water, and an appropriate amount of sodium hydroxide solution was added dropwise to neutralize the acrylic acid, with the degree of neutralization controlled at about 10%. The solution was stirred thoroughly and ultrasonically dispersed for 5 min to obtain the fourth solution. 2,2-diethoxyacetophenone (DEAP) was dissolved in dimethyl sulfoxide (DMSO) to prepare the fifth solution with a volume concentration of 10 v / v%. The fourth solution was placed in a refrigerator and refrigerated at 4°C for 24 hours to form hydrogen bonds and electrostatic complexes. After that, it was taken out and allowed to return to room temperature. Then, 50 µL of the fifth solution was added and stirred thoroughly to obtain the gel prepolymer. Nitrogen gas was then purged for deoxygenation before use.

[0081] (3) When using, use a 1 mL syringe to draw about 0.2 mL of the gel prepolymer solution prepared in step (2) above, and fill it into the photonic crystal interlayer of the molded container prepared in step (1) above. Note that air bubbles should not be introduced, and a small amount of gel prepolymer solution should be reserved at the edges. Then place the molded container in a UV crosslinker and use UV light to irradiate for 10 min to carry out the photopolymerization reaction to obtain the photonic crystal hydrogel. Soak the photonic crystal hydrogel in a centrifuge tube containing PBS buffer and place it on a shaker for later use.

[0082] (4) Methanol and acetic acid are mixed at a volume ratio of 9:1 to obtain a mixed solvent. A 1% sodium dodecyl sulfate solution is prepared. The mixed solvent and sodium dodecyl sulfate solution are mixed at a volume ratio of 1:1 and stirred thoroughly to obtain an eluent. The photonic crystal hydrogel prepared in step (3) above is immersed in the eluent and sonicated to elute the template molecules. The eluent is rotated periodically until creatinine is no longer detectable in the eluent. The elution time is about 6 hours, resulting in a functionalized photonic crystal hydrogel with a second specific recognition site for detecting creatinine.

[0083] The concentration prepared using PBS buffer at pH 7.4 was 10.0 mg / mL. 1 The creatinine standard solution was serially diluted to a concentration of 4.0 mg / mL. 1 2.0 mg mL 1 1.0 mg mL 1 0.5 mg mL 1 0.2 mg mL 1 0.1 mg mL 1 A diluted solution of creatinine.

[0084] The functionalized photonic crystal hydrogel prepared in Example 3 above was immersed in PBS buffer for 1 hour, and then... Figure 1 The testing apparatus shown was used to test the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different positions was taken; the functionalized photonic crystal hydrogels prepared in Example 3 above were respectively immersed in a solution with a concentration of 4.0 mg / mL 1 2.0 mg mL 1 1.0 mg mL 1 0.5 mg mL 1 0.2 mg mL 1 0.1 mg mL 1 In a diluted creatinine solution for 1 hour, use as follows Figure 1 The test setup shown tested the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different locations was taken. The test results are as follows: Figure 15 As shown.

[0085] from Figure 15 It can be seen that within the tested concentration range (0.1~4 mg / mL) 1 The interparticle spacing of the functionalized photonic crystal hydrogel decreased with increasing creatinine concentration, and the interparticle spacing of the functionalized photonic crystal hydrogel showed a good linear correlation with creatinine concentration (R0). 2 =0.96007). The limit of detection (LoD) was calculated to be 0.2692 mg / mL based on the ratio of the sample mean standard deviation (σ) to the slope of the calibration curve (S). 1 .

[0086] Example 4 This embodiment provides a functionalized photonic crystal hydrogel, the preparation method of which includes the following steps: (1) Preparation of the molded container: Five layers of tape are attached to the two short sides of the glass slide (first barrier layer) carrying the two-dimensional photonic crystal array obtained in Example 1 above. Another clean glass slide (second barrier layer) is used to sandwich the glass slide carrying the two-dimensional photonic crystal array (photonic crystal sandwich layer) to form a molded container with a "sandwich" structure.

[0087] (2) Preparation of gel prepolymer solution: Weigh 500 mg acrylamide (AM) into a 5 mL centrifuge tube, then weigh 15 mg N,N'-methylenebisacrylamide (MBA), add 50 mg acrylic acid (AA) and 2 mL H2O, sonicate to dissolve for 5 min, and finally add 30 µL 2,2-diethoxyacetophenone (DEAP), sonicate to obtain gel prepolymer solution for later use.

[0088] (3) When using, use a 1 mL syringe to draw about 0.2 mL of the gel prepolymer solution prepared in step (2) above, and fill it into the photonic crystal interlayer of the molded container prepared in step (1) above. Note that air bubbles should not be introduced, and a small amount of gel prepolymer solution should be reserved at the edges. Then place the molded container in a UV crosslinker and use UV light to irradiate for 18 min to carry out the photopolymerization reaction to obtain the photonic crystal hydrogel. Soak the photonic crystal hydrogel in a centrifuge tube containing PBS buffer and place it on a shaker for later use.

[0089] (4) Dissolve 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide in phosphate buffer at a molar ratio of 1:1 to prepare an activation solution with a concentration of 10 mM. Immerse the photonic crystal hydrogel prepared in step (3) above in the activation solution for 60 min to obtain the activated photonic crystal hydrogel.

[0090] (5) The activated photonic crystal hydrogels obtained in step (4) were immersed in urease solutions with concentrations of 0.5, 1, 2, and 3 mg / L prepared using PBS buffer, and incubated at 37 °C for 24 hours. Unbound enzyme proteins were then removed by washing with PBS buffer to obtain functionalized photonic crystal hydrogels with a third specific recognition site for urea detection. The functionalized photonic crystal hydrogels with different urease concentrations responded to 1 mmol / L... -1 The change in lattice spacing before and after urea production is as follows: Figure 16 As shown.

[0091] PBS buffer at pH 7.4 was used to prepare solutions with concentrations of 0.01, 0.1, 0.5, 1, 2, 4, 6, 8, and 10 mmol / L. -1 The urea solution. The functionalized photonic crystal hydrogel prepared in Example 4 above was soaked in PBS buffer for 1 hour, and then... Figure 1 The testing apparatus shown was used to test the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different positions was taken. The 2 mg / L urease-functionalized photonic crystal hydrogels prepared in Example 4 above were immersed in 0.01, 0.1, 0.5, 1, 2, 4, 6, 8, and 10 mmol L, respectively. -1In a urea solution for 1 hour, use as follows Figure 1 The test setup shown was used to measure the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different locations was taken. Linear fitting was performed on the lattice spacing of the functionalized photonic crystal hydrogels in response to different concentrations of urea, and the fitting results are shown below. Figure 17 As shown.

[0092] from Figure 17 It can be seen that within the tested concentration range (0.01-10 mmol L), -1 Within this range, the urea concentration is linearly correlated with the lattice spacing of the functionalized photonic crystal hydrogel (R0). 2 = 0.96583). The limit of detection (LoD) was calculated to be 93.1 µmol / L based on the ratio of the sample mean standard deviation (σ) to the slope of the calibration curve (S). -1 .

[0093] Example 5 This embodiment provides a functionalized photonic crystal hydrogel, the preparation method of which includes the following steps: (1) Preparation of the molded container: Five layers of tape are attached to the two short sides of the glass slide (first barrier layer) carrying the two-dimensional photonic crystal array obtained in Example 1 above. Another clean glass slide (second barrier layer) is used to sandwich the glass slide carrying the two-dimensional photonic crystal array (photonic crystal sandwich layer) to form a molded container with a "sandwich" structure.

[0094] (2) Preparation of gel prepolymer solution: Weigh 500 mg acrylamide (AM) into a 5 mL centrifuge tube, then weigh 15 mg N,N'-methylenebisacrylamide (MBA), add 50 mg acrylic acid (AA) and 2 mL H2O, sonicate to dissolve for 5 min, and finally add 30 µL 2,2-diethoxyacetophenone (DEAP), sonicate to obtain gel prepolymer solution for later use.

[0095] (3) When using, use a 1 mL syringe to draw about 0.2 mL of the gel prepolymer solution prepared in step (2) above, and fill it into the photonic crystal interlayer of the molded container prepared in step (1) above. Note that air bubbles should not be introduced, and a small amount of gel prepolymer solution should be reserved at the edges. Then place the molded container in a UV crosslinker and use UV light to irradiate for 18 min to carry out the photopolymerization reaction to obtain the photonic crystal hydrogel. Soak the photonic crystal hydrogel in a centrifuge tube containing PBS buffer and place it on a shaker for later use.

[0096] (4) Dissolve 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide in phosphate buffer at a molar ratio of 1:1 to prepare an activation solution with a concentration of 10 mM. Immerse the photonic crystal hydrogel prepared in step (3) above in the activation solution for 60 min to obtain the activated photonic crystal hydrogel.

[0097] (5) The activated photonic crystal hydrogels obtained in step (4) were immersed in anti-albumin antibody solutions with concentrations of 0.01, 0.03, 0.05, 0.1, and 0.3 mg / mL prepared using PBS buffer, and incubated at 37 °C for 24 hours. Unbound antibodies were then removed by washing with PBS buffer to obtain functionalized photonic crystal hydrogels with a fourth specific recognition site for albumin detection. The changes in lattice spacing before and after responding to 4 mg / mL albumin in functionalized photonic crystal hydrogels with different anti-albumin antibody concentrations are shown below. Figure 18 As shown.

[0098] Albumin solutions with concentrations of 1, 2, 3, 4, and 5 mg / mL were prepared using PBS buffer at pH 7.4. The functionalized photonic crystal hydrogel prepared in Example 5 was immersed in PBS buffer for 1 hour, and then... Figure 1 The testing apparatus shown was used to test the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different positions was taken. The 0.01 mg / mL anti-albumin antibody functionalized photonic crystal hydrogels prepared in Example 5 were immersed in albumin solutions of 1, 2, 3, 4, and 5 mg / mL for 1 hour, respectively, and then... Figure 1 The test setup shown was used to measure the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different locations was taken. Linear fitting was performed on the lattice spacing of different concentrations of albumin and the functionalized photonic crystal hydrogels in response to different concentrations of albumin; the fitting results are shown below. Figure 19 As shown.

[0099] from Figure 19 It can be seen that within the tested concentration range (1~5 mg / mL), the albumin concentration is linearly correlated with the lattice spacing of the functionalized photonic crystal hydrogel (R0). 2 =0.98318). The limit of detection (LoD) was calculated to be 0.384 mg / mL based on the ratio of the sample mean standard deviation (σ) to the slope of the calibration curve (S).

[0100] Example 6 Please see Figure 20The functionalized photonic crystal hydrogels prepared in Examples 2-5 above were integrated into different detection chambers in the same detection area of ​​the same detection chip, with each detection chamber having a volume of 0.5 cm³. 3 The chip casing is made of polydimethylsiloxane (PDMS).

[0101] Urine samples were serially diluted (1-fold, 5-fold, and 10-fold) using PBS buffer (pH 7.4) to obtain test solutions. The 10-fold diluted test solution was added dropwise to the inlet of the detection chip. The test solution flowed into each detection chamber through a microfluidic channel connected to the inlet, ensuring thorough immersion of the functionalized photonic crystal hydrogel within each chamber for 1 hour. Afterwards, the following steps were performed: Figure 1 The testing setup shown measures the diameter of the Debye diffraction rings of each group of functionalized photonic crystal hydrogels, and the average value of the test results at five different locations is taken. Based on the average diameter of the Debye diffraction rings, the quantitative concentrations of several markers, including glucose, creatinine, urea, and albumin, can be calculated.

[0102] The above method enables simultaneous detection of combined biomarkers (glucose + albumin) for diabetic nephropathy, allowing for disease early warning even when creatinine levels are not significantly elevated. This method detects disease abnormalities earlier than traditional single-indicator detection and is particularly suitable for postoperative home monitoring of patients or exercise metabolic monitoring of athletes.

[0103] The above embodiments are only used to illustrate the technical solutions of this application, and are not intended to limit them. Although this application has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of this application, and should all be included within the protection scope of this application.

Claims

1. A method for preparing a two-dimensional photonic crystal material, characterized in that, include: The polymer colloidal microspheres were uniformly dispersed in the first solvent to obtain a microsphere suspension; The microsphere suspension is mixed evenly with the second solvent to obtain a mixed solution; The mixed solution is slowly injected into a self-assembly container filled with ultrapure water, so that the mixed solution self-assembles on the liquid surface of the self-assembly container to form a two-dimensional photonic crystal array; The self-assembled container was transferred to a sealed gas chamber for organic solvent vapor annealing. Then, the two-dimensional photonic crystal array on the liquid surface of the container was transferred to a hydrophilically treated glass slide and dried to obtain a two-dimensional photonic crystal material. In the organic solvent vapor annealing step, the organic solvent saturated vapor used is any one or more of toluene saturated vapor, xylene saturated vapor, tetrahydrofuran saturated vapor, dichloromethane saturated vapor, or acetone saturated vapor.

2. The method for preparing a two-dimensional photonic crystal material according to claim 1, characterized in that, The first solvent is ultrapure water; the second solvent is n-propanol; the polymer colloidal microspheres are at least one of polystyrene microspheres or polymethyl methacrylate microspheres. The volume ratio of the microsphere suspension to the second solvent is 1:

1.

3. A two-dimensional photonic crystal material, characterized in that, The two-dimensional photonic crystal material is prepared by the preparation method of the two-dimensional photonic crystal material according to any one of claims 1 to 2.

4. A method for preparing a functionalized photonic crystal hydrogel, characterized in that, include: Preparation of gel prepolymer solution; The gel prepolymer is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light irradiation to carry out a photopolymerization reaction to obtain a functionalized photonic crystal hydrogel. The functionalized photonic crystal hydrogel has specific recognition sites for detecting disease biomarkers. The molding container includes a first barrier layer and a second barrier layer disposed at a relative interval; a photonic crystal interlayer is disposed between the first barrier layer and the second barrier layer; the photonic crystal interlayer includes the two-dimensional photonic crystal material as described in claim 3.

5. The method for preparing the photonic crystal hydrogel according to claim 4, characterized in that, The functionalized photonic crystal hydrogel has a first specific recognition site for glucose detection; Preparation of the gel prepolymer includes: Acrylamide and N,N'-methylenebisacrylamide were dissolved in ultrapure water to prepare the first solution; The second solution was prepared by dissolving 3-acrylamidophenylboronic acid in a third solvent; The photoinitiator was dissolved in a fourth solvent to prepare a third solution; The first solution, the second solution, and the third solution are mixed to obtain a gel prepolymer solution.

6. The method for preparing the photonic crystal hydrogel according to claim 4, characterized in that, The functionalized photonic crystal hydrogel has a second specific recognition site for detecting creatinine; Preparation of the gel prepolymer includes: Acrylic acid, hydroxyethyl methacrylate, N,N'-methylenebisacrylamide and creatinine were dissolved in ultrapure water, and sodium hydroxide solution was added and mixed evenly to obtain the fourth solution; The photoinitiator is dissolved in the fifth solvent to prepare the fifth solution; After refrigerating the fourth solution for 24 hours and then restoring it to room temperature, the fifth solution is added and mixed thoroughly to obtain a gel prepolymer solution. The gel prepolymer is filled into the photonic crystal interlayer of a molding container, and then subjected to photopolymerization under ultraviolet light irradiation to obtain a functionalized photonic crystal hydrogel, comprising: The gel prepolymer liquid is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light irradiation to carry out a photopolymerization reaction to obtain a photonic crystal hydrogel. A solution of methanol, acetic acid, and sodium dodecyl sulfate was mixed to obtain an eluent. The photonic crystal hydrogel was eluted multiple times using the eluent to obtain a functionalized photonic crystal hydrogel.

7. The method for preparing the photonic crystal hydrogel according to claim 4, characterized in that, The functionalized photonic crystal hydrogel has a third specific recognition site for detecting urea; Preparation of the gel prepolymer includes: Acrylamide, N,N'-methylenebisacrylamide and acrylic acid were dissolved in water, and then a photoinitiator was added and mixed evenly to obtain a gel prepolymer solution. The gel prepolymer is filled into the photonic crystal interlayer of a molding container, and then subjected to photopolymerization under ultraviolet light irradiation to obtain a functionalized photonic crystal hydrogel, comprising: The gel prepolymer liquid is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light irradiation to carry out a photopolymerization reaction to obtain a photonic crystal hydrogel. An activation solution was prepared by dissolving 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide in phosphate buffer. The photonic crystal hydrogel is placed in the activation solution for activation treatment to obtain the activated photonic crystal hydrogel. The activated photonic crystal hydrogel was incubated in a urease solution to obtain a functionalized photonic crystal hydrogel.

8. The method for preparing the photonic crystal hydrogel according to claim 4, characterized in that, The functionalized photonic crystal hydrogel has a fourth specific recognition site for detecting albumin; Preparation of the gel prepolymer includes: Acrylamide, N,N'-methylenebisacrylamide and acrylic acid were dissolved in water, and then a photoinitiator was added and mixed evenly to obtain a gel prepolymer solution. The gel prepolymer is filled into the photonic crystal interlayer of a molding container, and then subjected to photopolymerization under ultraviolet light irradiation to obtain a functionalized photonic crystal hydrogel, comprising: The gel prepolymer liquid is filled into the photonic crystal interlayer of the molding container, and then placed under ultraviolet light irradiation to carry out a photopolymerization reaction to obtain a photonic crystal hydrogel. An activation solution was prepared by dissolving 1-ethyl-(3-dimethylaminopropyl)carbodiimide and N-hydroxysuccinimide in phosphate buffer. The photonic crystal hydrogel is placed in the activation solution for activation treatment to obtain the activated photonic crystal hydrogel. The activated photonic crystal hydrogel was incubated in an anti-albumin antibody solution to obtain a functionalized photonic crystal hydrogel.

9. A functionalized photonic crystal hydrogel, characterized in that, The functionalized photonic crystal hydrogel is prepared by the preparation method of the functionalized photonic crystal hydrogel according to any one of claims 4 to 8.

10. A disease biomarker detection chip, characterized in that, The disease biomarker detection chip includes: Accommodating cavity; The receiving cavity is provided with at least one detection area, and each detection area includes a sample inlet and at least one detection chamber; the sample inlet is connected to each of the detection chambers through a microfluidic channel; each of the detection chambers is independent of each other and is not connected to each other; each of the detection areas is independent of each other and is not connected to each other. Each of the detection chambers includes the functionalized photonic crystal hydrogel prepared as claimed in claim 9.

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