179 results about "Fluorescent microspheres" patented technology

The invention relates to the technical field of detection kits, in particular to a preparation method of fluorescent microspheres, the fluorescent microspheres and a preparation method of a kit. According to the invention, 4-vinylbenzoic acid, dibenzoylmethane and phenanthroline are used as ligands to prepare the europium complex; the prepared europium complex, styrene and lauryl methacrylate are subjected to first-stage copolymerization polymerization to obtain a polymer core with the europium complex, itaconic acid is added for continuous polymerization in the second-stage polymerization process, and the prepared fluorescent microspheres are uniform in size, high in fluorescence intensity and high in stability. The prepared detection kit has the advantages of high detection sensitivity and stable and reliable detection effect when being used for detecting the myocardial injury marker cTnI.

The invention provides an immunodetection method of a multi-index immune checkpoint detection kit, which belongs to the technical field of immunodetection and comprises three main stages of sample preparation, immune reaction and data analysis. The method comprises the following steps: firstly, carrying out centrifugal separation to obtain a plasma sample, and preparing a fluorescent microsphere coupled capture antibody and biotin labeled detection antibody compound; then carrying out immune reaction, forming a sandwich type immune complex through specific binding, and adding streptavidin marked by phycoerythrin for signal amplification. Detecting a fluorescence signal by adopting a flow cytometer to obtain an original data matrix; a contribution feature vector, a utility feature vector and an expression feature matrix are extracted through a feature decomposition algorithm, and a deep learning model of a total-score-total structure is established for data analysis. Finally, a detection report containing a quantitative result, an importance score and reliability evaluation is generated, and the technical problem that in the prior art, the analysis accuracy of a multi-index immune checkpoint detection result is insufficient is solved.

The invention provides a fracture deformation-seepage cross-scale visualization experiment device and a test method, and relates to the technical field of hydraulic rock mechanics and measurement. Comprising a macroscopic observation assembly, a microscopic observation assembly, a microscopic seepage model, a temperature-stress-seepage loading system, a monochromatic coaxial light source and a data acquisition and analysis system, the microscopic seepage model is used for placing a fractured rock mass; the temperature-stress-seepage loading system is used for injecting fluid with fluorescent microspheres into the microcosmic seepage model, applying load, regulating and controlling temperature and collecting data; the data acquisition and analysis system is used for processing data and images. By arranging the macroscopic observation assembly and the microcosmic observation assembly, double-scale synchronous observation is achieved, the whole process of fracture internal fluid interface evolution and rock mass deformation can be obtained in real time under the multi-physics field coupling conditions of temperature, stress, seepage and the like, visual correlation of deformation, flow and stress is achieved, and the measurement accuracy is improved. And an experimental basis is provided for establishing a fracture multi-field coupling mechanism model.

The invention discloses a kit for jointly detecting phosphorylated Tau-217 and non-phosphorylated Tau-217. The kit comprises a magnetic microsphere reagent, a detection antibody reagent and a fluorescent microsphere reagent, the magnetic microsphere reagent comprises a magnetic microsphere compound, the detection antibody reagent comprises a first detection antibody compound and a second detection antibody compound, and the fluorescent microsphere reagent comprises a first fluorescent microsphere compound and a second fluorescent microsphere compound. The invention provides a kit for jointly detecting phosphorylated Tau-217 and non-phosphorylated Tau-217, which is based on a single molecule detection technology, and realizes accurate distinguishing and high-sensitivity detection on the phosphorylated Tau-217 and the non-phosphorylated Tau-217 at the same time through a double-target synchronous detection design and a double-color fluorescence report system.

The invention discloses a method for simulating a spreading rule of African swine fever virus aerosol in a pig farm, and relates to the technical field of animal epidemic disease spreading simulation. Comprising the following steps: selecting fluorescent microspheres, and preparing a fluorescent microsphere suspension; building a fluorescent microsphere suspension release system; dividing a pig farm into a plurality of cells, and arranging a sampling network in the pig farm; starting all the samplers of the sampling network, idling for a certain time, and closing all the samplers after measuring the environment background concentration; synchronously starting a fluorescent microsphere suspension release system and samplers of all sampling networks, and synchronously recording environment temperature and humidity, wind speed and wind direction; and performing quantitative analysis on the fluorescence of the pig farm at different time points, and generating a distribution diagram and a dynamic change diagram of the fluorescence in the pig farm according to an analysis result. According to the invention, by releasing the fluorescent microspheres with matched particle sizes and combining with a gridding monitoring network, a distribution diagram and a dynamic change diagram during virus diffusion are dynamically generated, and data support is provided for pig farm ventilation optimization and accurate disinfection.

The invention belongs to the technical field of biological detection, and provides a multiple cell factor detection kit in tumor immunity and a preparation method and application thereof, and the detection kit comprises a capture antibody, a detection antibody, a mixed protein standard substance, a diluent and a washing buffer solution. The capture antibody is prepared by coupling a biotinylated antibody with a fluorescent microsphere coated with biotin to obtain a capture antibody coupled with a fluorescent encoding microsphere, and the detection antibody is prepared by coupling derivatized phycoerythrin PE-SMCC with a specific thioether bond of a sulfhydrylated antibody to obtain a fluorescent labeled antibody; the mixed protein standard substance adopts a freeze-drying protection system containing BSA, trehalose and the like. The single hole of the kit can synchronously detect various cell factors, the sample dosage is only 25 microliters, the kit is suitable for immune monitoring of iron overload and other chronic inflammation models, compared with a traditional ELISA method, the efficiency is improved by 10 times, the cost is reduced, and the kit has the remarkable advantages of being high in throughput, wide in linear range and high in stability.

The invention discloses a preparation method and application of an HPV16 / 18 type E6 / E7 protein multi-target detection test strip, and belongs to the technical field of biological immunodetection analysis. The detection test strip comprises a back plate, a sample pad, a combination pad, a nitrocellulose membrane and an absorption pad, AIE fluorescent microsphere labeled probes are distributed on the combination pad, and a capture antibody and an AIE solution are distributed on the nitrocellulose membrane. The preparation method comprises the following three steps: preparation of the nitrocellulose membrane, preparation of the combination pad and assembly of the test strip. The detection test strip can synchronously and quantitatively detect PV16 E6, HPV16 E7, HPV18 E6 and HPV18 E7, each target corresponds to four signal points, multi-target synchronous rapid detection is achieved, the linear range is wide, the detection limit is low, operation is easy and convenient, cost is low, and the detection test strip is expected to become a novel efficient and reliable on-site screening tool for cervical cancer and precancerous lesions.

The invention relates to the technical field of immunochromatography, in particular to time-resolved fluorescent microspheres, a preparation method and application of the time-resolved fluorescent microspheres in PSP / CRP / MxA quantitative detection immunochromatography test strip, and the preparation method of the time-resolved fluorescent microspheres comprises the following steps: 1) preparing surface aminated polystyrene microspheres; (2) carrying out glutaraldehyde crosslinking and 1, 10-phenanthroline-5-amino combination; 3) grafting a rare earth fluorescent dye on the surface; and 4) coating a polymer shell layer and modifying carboxyl. According to the invention, through a dual fixation strategy of surface grafting and polymer coating, the time-resolved fluorescent dye is confined to a functional region between the surface of the microsphere and a polymer shell layer, diffusion and leakage of dye molecules to the external environment are blocked, and the problem of dye shedding of traditional fluorescent microspheres is solved; the polymer shell layer can effectively isolate external ultraviolet light, oxygen and other corrosive factors, reduce photooxidative degradation and non-radiative transition loss of fluorescent molecules, and improve the light stability and fluorescent lifetime of the fluorescent molecules.

The invention provides a flow cytometer light path calibration method, which belongs to the technical field of flow cytometers, and comprises the following steps: firstly, collecting light path detection original data of standard fluorescent microspheres, including laser spot position coordinates, detector output voltage value, dark counting rate and signal counting rate; then establishing a detection channel conversion cost matrix, and calculating an optimal light path detection sequence by using a traveling salesman problem optimization algorithm; adjusting the positions of a laser, an optical filter and a detector according to the optimal sequence, and obtaining optical path calibration data; the collected parameters are substituted into an optical path parameter optimization equation set to be solved, and an optical path collimation coefficient, a detector response coefficient, a signal crosstalk coefficient and an optimal gain value are obtained; and finally, respectively adjusting the position of the laser, the working voltage of the detector, the position of the optical filter and the voltage of the photomultiplier according to the coefficients to finish the accurate calibration of the optical path. The problems that an existing method depending on artificial experience is low in efficiency and overall optimization of a system level is difficult to achieve are solved.

The invention belongs to the technical field of biological detection, and provides a multiple cell factor detection kit in macrophage polarization and a preparation method and application thereof, and the detection kit comprises a capture antibody, a detection antibody, a mixed protein standard substance, a diluent and a washing buffer solution. The capture system is used for preparing a capture antibody coupled with a fluorescence coding microsphere by coupling a biotinylated antibody with a fluorescence microsphere coated with biotin, and the detection system is used for preparing a fluorescence labeled antibody by coupling derivatized phycoerythrin PE-SMCC with a specific thioether bond of a sulfhydrylated antibody. The mixed protein standard substance adopts a freeze-drying protection system containing BSA, trehalose and the like. The single hole of the kit can synchronously detect multiple cell factors, the sample dosage is only 25 microliters, the kit is suitable for in-vitro tests of mouse mononuclear macrophage leukemia cell lines RAW264.7, compared with a traditional ELISA method, the efficiency is improved by 10 times, the cost is reduced, and the kit has the remarkable advantages of being high in throughput, wide in linear range and high in stability.

The invention discloses a preparation method and application of high-sensitivity and high-stability time-resolved fluorescent microspheres, and relates to the technical field of immunochromatography. The preparation method comprises the following steps: (1) preparing high-stability high-carboxyl-density polystyrene microspheres; (2) preparing a high-stability europium ion chelate; and (3) preparing the high-stability fluorescent microspheres. Through a strategy of combining functional monomer copolymerization and surface covalent labeling, the fluorescent microspheres which are connected through strong covalent bonds from inside to outside and are high in stability and carboxyl density are constructed; obvious differential advantages and innovativeness are shown in the aspects of chemical stability, anti-leakage capability of fluorescent markers and surface functional group density. A time-resolved fluorescent microsphere test strip developed based on the microspheres has the advantages of good stability, high detection sensitivity and the like, and can realize rapid detection and analysis of a substance to be detected.

The present application relates to the technical field of microfluidic chip, and disclose a centrifugal microfluidic chip for generating different size droplets simultaneously, including a liquid distribution channel for uniformly distributing sample to multiple droplet generation units, the design of a quantitative pool ensures the fixed volume of liquid and the consistency of the number of droplet generation, the chip contains a sample inlet and two continuous phase reagent inlets, the sample is precisely distributed to the quantitative pool by centrifugal force and enters the droplet generation units respectively, the first droplet generation unit is used for generating large size droplets with a diameter of 100-200 mu m, and the second droplet generation unit is used for generating small size droplets with a diameter of 10-100 mu m, the microfluidic chip can realize the preparation of ultrahigh throughput droplets, the generated droplets are guided to the collection area by centrifugal force to form a regular single-layer arrangement, the chip has wide application in the fields of biomedical research, material science, chemical synthesis and environmental monitoring, especially in the preparation of fluorescent microspheres, which can improve the experimental efficiency and result consistency.

The application discloses a detection method of cell surface antigens and application thereof, and relates to the detection field. The detection method of cell surface antigens comprises the following steps: mixing liposome-treated cells to be detected with capture microspheres and fluorescent microspheres, detecting the fluorescence intensity, and analyzing the expression of cell surface antigens. The surface of the capture microspheres is coated with a first antibody against a specific antigen on the surface of target cells. The surface of the fluorescent microspheres is coated with a second antibody against a target antigen on the surface of target cells. The detection method has low cost and can be used for directly quantitatively and qualitatively analyzing the expression of surface antigens of specific types of cells.

The invention discloses a biomarker combination related to hypoxia-induced neuroinflammation and application of the biomarker combination, and belongs to the technical field of biomarker detection. The marker combination disclosed by the invention consists of serum S100 beta protein, neuron-specific enolase, interleukin-6, high-mobility group protein B1 and neurofilament light-chain protein. The marker combination can be used for stratification of nervous system injury risk of severe hypoxia patients and acquisition of prognosis evaluation data. According to the detection method disclosed by the invention, synchronous quantitative detection of five markers is realized by adopting a multiple immunofluorescent microsphere technology, and a marker combination scoring algorithm is established to comprehensively evaluate the severity of the hypoxia-related neuroinflammation.

The present invention relates to the field of tracer technology, and discloses a codable and decodable magnetic fluorescent tracer, comprising a magnetic fluorescent microsphere comprising a core formed by a plurality of fluorescent materials, a polymer coating the core, and a plurality of magnetic microspheres embedded on the surface of the polymer. The tracer has both magnetic and fluorescent functions. When sampling and testing production wells, the tracer can be enriched using a magnetic field, reducing the difficulty of purification from the produced fluid. Compared with direct testing of the original fluid, the error of subsequent analysis and detection after enrichment of the tracer is significantly reduced. The tracer can be prepared in more than 20 different colors and can be decoded using a flow cytometer, increasing the variety of fluorescent tracers.

The application discloses a lateral test strip for detecting C-reactive protein and application thereof, and the test strip comprises a bottom plate and a sample pad, a combination pad, a nitrocellulose membrane and a water absorption paper which are sequentially stacked on the bottom plate; the combination pad is loaded with AIE fluorescent microspheres; the AIE fluorescent microspheres are composed of P(St-MMA) microspheres coated with AIE materials after hydrolysis and carboxylation; and anti-C-reactive protein antibodies are connected to the AIE fluorescent microspheres through a covalent bond. The surface coating amount of the AIE fluorescent microspheres is increased by 1.6-2.3 times, and the fluorescence intensity per unit mass is increased by about 40-60%. The increase of the coating amount makes the density of the light-emitting core higher, the overall quantum yield is significantly improved, and the uniformity of the fluorescence signal and the detection sensitivity are enhanced. The characteristics are particularly prominent in the detection of CRP protein in the chromatographic test strip, and the detection limit can be effectively reduced, the T / C signal ratio can be improved, and the reaction time can be shortened.

The invention relates to the technical field of tracers, and discloses a fluorescent microsphere, a tracer and a preparation method thereof, the fluorescent microsphere comprises a core formed by coating a fluorescent material with a polymer, a first shell layer coating the core and a second shell layer coating the first shell layer, and the second shell layer is connected with the first shell layer through a covalent bond. When the fluorescent microspheres are prepared, the polymer fluorescent microspheres are coated with the first shell layer in a physical adsorption mode, the first shell layer is coated with the second shell layer in a chemical bond connection mode, and the second shell layer is smoother, more compact and firmer in connection due to the chemical bond connection mode, so that the shell layers cannot fall off, and the stability of the fluorescent microspheres is effectively improved.

The application discloses a labeling method capable of improving fluorescence amplification effect and application thereof in biological toxin detection, and the labeling method comprises the following steps: S1, double-fluorescent microsphere preparation: S1.1, performing carboxyl site blocking on aflatoxin B1, vomitoxin and zearalenone antibodies; S1.2, performing coupling on amino sites of the closed aflatoxin B1, vomitoxin and zearalenone antibodies; S1.3, activating the coupled aflatoxin B1, vomitoxin and zearalenone antibodies; and S1.4, coupling the activated aflatoxin B1, vomitoxin and zearalenone antibodies with magnetic fluorescent microspheres. The labeling method capable of improving fluorescence amplification effect and application thereof in biological toxin detection have the advantages that the sensitivity and detection limit of products can be improved, and the detection difference caused by the interference of sample matrix can be reduced.

This invention relates to the field of biodetection technology, and more particularly to a fluorescent test strip for detecting canine parvovirus, along with hybridoma cells, monoclonal antibodies, and their applications. The test strip of this invention has fluorescent microspheres coated with canine parvovirus monoclonal antibody and fluorescently labeled rabbit IgG within the conjugate pad; a detection line and a control line are disposed on the surface of a nitrocellulose membrane, with the detection line coated with canine parvovirus monoclonal antibody and the control line coated with rabbit anti-IgG; wherein the canine parvovirus monoclonal antibody is secreted using hybridoma cells with accession number C202355. The test strip of this invention can be used to simultaneously detect feline parvovirus, canine parvovirus, and various genotypes of canine parvovirus, exhibiting high sensitivity and specificity, and possessing the technical advantages of accuracy, speed, and simplicity.

The present invention provides a latex microsphere conjugate or a fluorescent microsphere conjugate, comprising an antibody and a conjugate moiety, wherein 1-27 amino acid residues are chimerically inserted after the C-terminal lysine residue of the heavy chain of the antibody. The conjugate has high activity or high sensitivity.

The invention relates to a synchronous detection method and system for small molecules and protein, and the synchronous detection method comprises the following steps: respectively preparing a small molecule detection antibody and a protein detection antibody marked by carboxyl fluorescent microspheres, and mixing the small molecule detection antibody and the protein detection antibody to obtain a mixed detection antibody; respectively preparing a microsphere-labeled capture micromolecule antigen and a protein antibody; preparing a microsphere array biochip with two detection areas; carrying out sample application on the microsphere-labeled captured micromolecule antigen and the protein antibody to the two detection areas; mixing a sample to be detected with the mixed detection antibody according to different proportions; slowly adding the sample mixed solution into two detection areas of the microsphere array biochip respectively; scanning and imaging the two detection areas of the microsphere array biochip respectively, identifying all fluorescent spot information in a fluorescent image, and obtaining the concentration of small molecules and proteins in a sample to be detected through a Poisson distribution principle or by establishing a standard curve. According to the invention, synchronous detection of small molecules and proteins can be realized.

The utility model discloses a fluorescent quantitative rapid detection card for ofloxacin quantum dot microspheres. The ofloxacin quantum dot microsphere fluorescent quantitative rapid detection card comprises a test strip, the test strip comprises a bottom plate, a sample pad, a fluorescent microsphere conjugate pad, a nitrocellulose membrane and a water absorption pad, the sample pad, the fluorescent microsphere conjugate pad, the nitrocellulose membrane and the water absorption pad are arranged on the bottom plate and connected in sequence, and detection lines and quality control lines which are distributed at intervals are arranged on the nitrocellulose membrane. The detection line is close to the fluorescent microsphere conjugate pad, the quality control line is close to the water absorption pad, the detection line is coated with a micromolecule ofloxacin antigen, the quality control line is coated with a goat anti-mouse IgG antibody, and a microsphere labeled antibody conjugate in the fluorescent microsphere conjugate pad is a fluorescent microsphere labeled ofloxacin antibody. The method is simple and rapid to operate, is less affected by environment temperature and solvent reagents, is suitable for rapid detection of food safety, and is accurate in detection result.

The application discloses a high-brightness quantum dot fluorescent microsphere and a preparation method thereof based on an alkali catalysis strategy, and comprises the following steps: (1) preparing thiol-modified dendritic mesoporous silica microspheres; (2) assembling the thiol-modified dendritic mesoporous silica microspheres and quantum dots to obtain a dendritic silica / quantum dot assembly; (3) adding 3-mercaptopropyl triethoxysilane to the dendritic silica / quantum dot assembly, and performing organic silane modification on the dendritic silica / quantum dot assembly in a non-polar organic solvent by taking an organic alkali as an alkali catalyst; and (4) growing a silica shell layer on the surface of the organic silane-modified dendritic silica / quantum dot assembly by a method to obtain the high-brightness quantum dot fluorescent microsphere. The preparation method avoids fluorescence quenching of oil-phase quantum dots in the process of coating silica, and completely retains excellent optical properties of the oil-phase quantum dots.

The invention provides a low-abundance antigen detection method based on flow-type fluorescent single molecule counting, and belongs to the technical field of biological detection.The method comprises the following steps that after fluorescent encoding microspheres coated with captured antibodies are mixed with a standard sample or a sample to be detected, detection antibodies and nano magnetic beads are added, and the fluorescent encoding microspheres are obtained; or directly adding the nano magnetic beads coated with the detection antibody, reacting to form a double-antibody sandwich compound, separating the compound, and counting by adopting a flow cytometer; and establishing a standard curve of the antigen concentration and the collection number of the fluorescent microspheres, and calculating the concentration of the target antigen in the to-be-detected sample according to the standard curve. The method effectively improves the problem of low sensitivity of a flow fluorescence technology, is low in detection cost and reliable in detection result, and has important significance on low-abundance antigen detection.

The invention provides a progesterone detection reagent strip based on an antibody synergistic effect and a preparation method of the progesterone detection reagent strip. The progesterone detection reagent strip comprises a bottom plate, a sample pad, a combination pad, a nitrocellulose membrane and a water absorption pad, the combination pad is sprayed with fluorescent nano-microspheres coupled with a goat-derived PROG antibody and fluorescent nano-microspheres coupled with a goat anti-rabbit antibody, a detection line of the nitrocellulose membrane is coated with a PROG antigen, and a quality control line of the nitrocellulose membrane is coated with a rabbit anti-goat IgG antibody. According to the progesterone detection reagent strip based on the antibody synergistic effect, the competition method detection principle is adopted, the fluorescent microspheres are used as markers, the progesterone antibody and the fluorescent microspheres are combined in a coupling mode, the specificity and binding efficiency of antigen-antibody reaction are optimized based on the antibody synergistic effect, and the progesterone detection reagent strip can be used for detecting progesterone. The stability and sensitivity of the detection system are remarkably improved, and high-sensitivity and high-specificity detection of the progesterone level in serum or plasma is achieved.

The application discloses a preparation method and application of an HPV16 / 18 type E6 / E7 protein multi-target detection test strip, and belongs to the technical field of biological immune detection and analysis. The test strip comprises a back plate, a sample pad, a combination pad, a nitrocellulose membrane and an absorption pad, AIE fluorescent microsphere labeled probes are distributed on the combination pad, and capture antibodies and AIE solution are distributed on the nitrocellulose membrane. The preparation method comprises three steps of preparation of the nitrocellulose membrane, preparation of the combination pad and assembly of the test strip. The test strip can quantitatively detect PV16 E6, HPV16 E7, HPV18 E6 and HPV18 E7 simultaneously, four signal points correspond to each target, multi-target synchronous rapid detection is realized, the linear range is wide, the detection limit is low, the operation is simple and convenient, the cost is low, and the test strip is expected to become a new tool for efficient and reliable on-site screening of cervical cancer and precancerous lesions.

The invention discloses a preparation method and application of a clothianidin fluorescent test strip, belongs to the technical field of agricultural detection, and solves the problems that the existing bombyx mori pesticide poisoning detection sensitivity is low, and the clothianidin residue concentration cannot be quickly judged. According to the method, a nitrocellulose membrane is prepared, a detection line is coated with a clothianidin-BSA antigen, and a quality control line is coated with an anti-mouse IgG antibody; when the fluorescent pad is prepared, carboxylated fluorescent microspheres and an anti-clothianidin antibody are coupled to form a labeled probe; when the test strip is assembled, the nitrocellulose membrane, the fluorescent pad, the sample pad and the absorbent paper are stacked and fixed on the bottom plate to form a chromatography structure. Clothianidin in a sample is combined with the labeled probe to form an immune complex, chromatography migrates to the detection line to be competitively combined with clothianidin-BSA antigen, the uncombined probe is captured and develops color, and the quality control line develops color through an anti-mouse IgG antibody. The test strip is used for rapidly detecting clothianidin residues in mulberry leaves and bombyx mori bodies, the sensitivity reaches 0.02 mg / L, pesticide poisoning events can be effectively prevented, and safe production of the sericulture industry is guaranteed.

The invention discloses a fluorescence immunodetection analyzer and a detection analysis method thereof. The analyzer comprises a sheath flow pool which is vertically arranged, red and green laser excitation light sources which are horizontally arranged on the two sides of the sheath flow pool in the X-axis direction, and double independent light paths which are arranged on the two sides of the sheath flow pool in the Y-axis direction. A red laser light path separates fluorescence into three paths of signals of 730nm, 670nm and residual fluorescence through a series-connected dichotomy mirror group; the green laser light path collects diffuse reflection light. The four paths of optical signals are converted into voltage signals by four detectors, temperature compensation units integrated in the detectors correct the signals in real time based on a pre-calibrated dark current-temperature relation curve, and temperature drift is suppressed. And the signal processing system calls a pre-established signal model library to carry out comprehensive calibration and synchronous analysis on multi-channel signals, so that accurate distinguishing and counting of different fluorescent microspheres are realized. According to the invention, the problems of large temperature influence and low detection efficiency in the prior art are solved, and the stability, accuracy and multi-channel consistency of detection are remarkably improved.

The invention relates to the field of protein detection, in particular to a method for detecting phosphorylated Tau-217 protein in urine of an Alzheimer's disease patient, which comprises the following steps of: 1, collecting fresh urine in the morning by using a urine collecting container, and 2, taking out 100 microliters of urine by using a transfer pipette and adding the urine into a 5.0 ml centrifugal tube; step 3, adding 4.9 ml of a urine diluent to dilute the urine; and step 4, prefabricating tracing fluorescent microspheres and enrichment microspheres in the centrifugal tube. Step 5, repeatedly reversing and uniformly mixing, and reacting at room temperature for 8-12 hours; step 5, placing the reaction tube on a separation test tube rack with a magnet for more than 20 minutes; step 6, irradiating the test tube by using a highlighter with the wavelength of about 340nm in a dark environment to judge a result; and step 7, if a red fluorescent dot appears near the magnet in the test tube, the result is positive, otherwise, the result is negative, and the result can also be judged by measuring a fluorescence value through a luminoscope with the wavelength of 340nm. Finally, the phosphorylated Tau-217 protein is detected, and the operation threshold and the economic cost are reduced.

The invention discloses a large-scale controllable preparation method of super-bright fluorescent silicon microspheres based on fluorescent probe doping, and belongs to the technical field of nano material preparation. According to the method, firstly, a fluorescent probe is efficiently embedded into a silicon skeleton, so that a microsphere fluorescent signal is enhanced, a raindrop effect is avoided, and the problems of low brightness and non-uniform luminescence of a traditional fluorescent microsphere are solved; through multi-step refined modification, the interface compatibility of the fluorescent probe and a silica sol system is improved, and uniform doping and stable embedding are ensured; the fluorescent probe is doped into the silicon microspheres at high density, so that the unit signal intensity and the imaging consistency are improved, and the problem of outer surface modification is avoided. According to the preparation process, double injection pumps are adopted for synchronous feeding, a large-volume reaction system and constant-temperature stirring are adopted, 0.5-3 g of dry microspheres can be stably prepared in a single batch, and the preparation efficiency is improved. The prepared superbright fluorescent silicon microspheres have the advantages of high brightness, high stability, good dispersity, batch consistency and the like, and a feasible scheme is provided for batch production of fluorescent coding probes.