311 results about "Fluorescent microspheres" patented technology

The invention relates to the technical field of biological detection, and provides an inflammatory factor detection kit based on a liquid-based chip technology, and the inflammatory factor detection kit comprises six fluorescent microspheres coupled with capture antibodies, and six detection antibodies labeled by report fluorescence; in the six fluorescent microspheres containing the captured antibodies, the fluorescence intensities of all the fluorescent microspheres are different; according to the detection kit provided by the invention, on the basis of a liquid-based chip technology, fluorescein is adopted to carry out fluorescence coding on microspheres, and a captured antibody is connected to the fluorescent microspheres through a chemical reaction; a detection target is captured through the fluorescent microspheres, and then a signal is displayed through the detection antibody, so that the detection of the inflammatory factors is completed, the defects of large demand quantity of soluble inflammatory factor detection samples, long detection time, tedious detection steps and the like in the prior art are overcome, and great convenience is brought to scientific researchers at a hospital end and an industrial end.

The invention discloses a test strip for five-item combined detection of inflammation of peripheral blood as well as a preparation method and application of the test strip. The test strip comprises a PVC bottom plate, a nitrocellulose membrane, a glass cellulose membrane combination pad, a sample pad and a water absorption pad, the PVC bottom plate is located at the bottommost part, and the sample pad, the glass cellulose membrane combination pad, the nitrocellulose membrane and the water absorption pad are sequentially arranged above the PVC bottom plate; the nitrocellulose membrane is provided with an HBP, PCT, IL-6, CRP and SAA detection line and a mouse anti-DNP monoclonal antibody quality control line; the glass nitrocellulose membrane combination pad is provided with a combination of a fluorescent microsphere-labeled HBP antibody, a fluorescent microsphere-labeled PCT antibody, a fluorescent microsphere-labeled IL-6 antibody, a fluorescent microsphere-labeled CRP antibody, a fluorescent microsphere-labeled SAA antibody and a fluorescent microsphere-labeled DNP-BSA. According to the test strip, fluorescence immunochromatography is carried out by utilizing the principle of a double-antibody sandwich method in combination with the characteristics of fluorescent microspheres, and rapid, specific, simple, convenient and sensitive joint detection on HBP, PCT, IL-6, CRP and SAA is realized at the same time.

The invention discloses a multi-fluorescent-channel microsphere image alignment method, device and equipment. The method comprises the following steps: respectively acquiring fluorescent microsphere images at the same position through multiple fluorescent channels; preprocessing each fluorescent microsphere image to obtain a fluorescent correction image; performing microsphere region segmentation on each fluorescence correction image, and calculating a region center point corresponding to each microsphere region; intercepting a pixel matrix from the fluorescence correction image according to the central point of each region, and extracting a corresponding feature vector; any one fluorescence correction image is selected as a reference image, the remaining fluorescence correction images are subjected to pairwise registration according to the feature vectors, and adjustment offset is determined respectively; and correcting each fluorescence correction image according to the adjustment offset, and superposing the corrected fluorescence correction images to the reference image to generate a fluorescence channel alignment image. Therefore, high-precision image alignment is realized without depending on deep learning in a mode of performing offset registration on the fluorescent microsphere image in combination with the gradient direction.

The invention relates to the technical field of biological detection, in particular to an MxA, CRP and SAA combined detection card and a preparation method and application thereof, and the preparation method comprises the following steps: 1) preparing silicon-coated high-performance time-resolved fluorescent microspheres; (2) marking of an MxA antibody; (3) marking a CRP antibody; (4) marking an SAA antibody; (5) preparing a conjugate pad; (6) preparing a nitrocellulose membrane; and (7) assembling the reagent strip. The silicon-coated high-performance time-resolved fluorescent microspheres with the particle size range of 190-352 nm are prepared, the dye embedding rate is high, the sensitivity is high, the monodispersity is good, the stability is good, the hydrophilicity is high, agglomeration is not prone to occurring, and the dye embedding rate is increased. The MxA, CRP and SAA combined detection card prepared from the silicon-coated high-performance time-resolved fluorescent microspheres can improve the accuracy of identifying bacterial infection and virus infection, and is beneficial to early diagnosis and treatment of acute respiratory infectious diseases.

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 discloses a micro-plastic integrated intelligent detection device, which comprises: a sample pretreatment module, which comprises a sample mixing module, a filtering separation module, an enrichment concentration module and a surface modification and marking module which are communicated in sequence and is used for mixing, filtering, enriching, modifying and marking a sample in sequence; the immunoreaction module is used for performing immunoreaction on the sample and the anti-polystyrene monoclonal antibody freeze-dried product marked by the quantum dot fluorescent microspheres; the fluorescence detection module is used for carrying out fluorescence detection on the test strip subjected to the immune reaction; the automatic control module is electrically connected with the sample pretreatment module, the immunoreaction module and the fluorescence detection module and is used for controlling the operation of each module. According to the invention, the sample processing module, the reaction module, the detection module and the control module are in close cooperation, the sample enters each module in sequence after simple pretreatment, detection and result output are semi-automatically completed, frequent manual operation of different instruments is not needed, the operation steps and time are reduced, and the operation difficulty is reduced.

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 discloses a polystyrene micro-nano plastic immunochromatography time-resolved fluorescence kit and application thereof.The kit comprises a fluorescent test strip and a sample reaction bottle containing a quantum dot fluorescent microsphere labeled anti-polystyrene monoclonal antibody freeze-dried product, and the fluorescent test strip comprises a bottom plate; a water absorption pad, a detection pad and a sample pad are sequentially adhered to the adhesive surface of the bottom plate from top to bottom, adjacent pads are overlapped and connected at joints, a nitrocellulose membrane serves as a base pad of the detection pad, a transverse quality control line and a detection line are arranged on the nitrocellulose membrane from top to bottom, the quality control line is coated with an anti-mouse-free polyclonal antibody, and the detection line is coated with an anti-mouse-free polyclonal antibody. And the detection line is coated with a polystyrene-hemocyanin conjugate. The immunochromatography time-resolved fluorescence kit provided by the invention can realize quantitative detection of polystyrene, and has the advantages of strong specificity, high sensitivity, simple sample pretreatment, small pollution hazard to the environment, suitability for on-site batch detection and the like.

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 provides monoclonal antibodies of H protein and N protein of canine distemper virus and application thereof in preparation of a fluorescent microsphere antigen detection test strip. The monoclonal antibody disclosed by the invention is characterized in that (1) the monoclonal antibody contains a heavy chain variable region CDV-N-3A11-VH and a light chain variable region CDV-N-3A11-VL; the amino acid sequence of the CDV-N-3A11-VH is as shown in the 1st to 116th sites of SEQ ID No.1 in a sequence table; the amino acid sequence of the CDV-N-3A11-VL is as shown in the 1st site to the 113th site of SEQ ID No.2 in a sequence table. (2) a heavy chain variable region CDV-H-1G5-VH and a light chain variable region CDV-H-1G5-VL are contained; the amino acid sequence of the CDV-H-1G5-VH is as shown in the first site to the 114 site of SEQ ID No.3 in a sequence table; the amino acid sequence of the CDV-H-1G5-VL is as shown in the 1st site to the 1st site and the 1st site to the 1st site of SEQ ID No.4 in a sequence table. The canine distemper virus fluorescent microsphere antigen test strip composed of the two antibodies is suitable for detecting canine distemper viruses in samples such as eye and nose swabs and virus cultures, is high in sensitivity and good in specificity, and can be used for early diagnosis of canine distemper virus infection.

The invention relates to the technical field of microimaging automatic focusing, in particular to an automatic cell focusing method and system capable of resisting interference of fluorescent microspheres in hydrogel. The method comprises the following steps: acquiring hydrogel cell image sets containing fluorescent microspheres on different focal planes, calculating cell region weights and weighted gradient processing according to the hydrogel cell image sets containing fluorescent microspheres on different focal planes, and storing the cell region weights and weighted gradient processing to a preset definition list; and traversing the definition list, performing definition scoring on the hydrogel cell images containing the fluorescent microspheres on different focal planes in the definition list to obtain the hydrogel cell image containing the fluorescent microspheres with the highest definition score and the corresponding Z-axis coordinate, and controlling a preset motor to realize automatic focusing by using the Z-axis coordinate. The method can adapt to various fluorescent microsphere densities, is not influenced by the densities, and has wide applicability; the algorithm considers the existence of all cells in the visual field, the focusing is more integral, and the focusing accuracy is improved.

The invention relates to the technical field of immunochromatography, in particular to a silicon-coated color fluorescent microsphere and a mild preparation method and application thereof.The preparation method comprises the steps that firstly, time-resolved fluorescent dye is synthesized in an ethanol system, then oil-soluble dye, an ethanol water solution and a swelling agent dichloromethane are added, stirring is conducted at the room temperature to enable the dye to be dissolved, and then the silicon-coated color fluorescent microsphere is obtained; and adding ammonia water and a silane coupling agent tetraethyl orthosilicate, reacting overnight, supplementing the ammonia water, gradually adding a coupling agent 3-aminopropyltriethoxysilane, and washing with pure water for several times after the reaction is finished. The particle size range of the prepared color time-resolved fluorescent microspheres is 200-450 nm, the color time-resolved fluorescent microspheres are prepared in a green and mild mode through a one-step method, a surface active agent, a dispersing agent and a surface amino modified silicon dioxide matrix are not used, the high-luminous-intensity binuclear rare earth europium complex and the oil-soluble dye are embedded into the microspheres in the polymerization process, the dye is not prone to leakage, and the color time-resolved fluorescent microspheres are environmentally friendly and environmentally friendly. The method can be used for qualitative and quantitative dual immunodetection in the field of immunochromatography.

The invention discloses a method and a device for detecting errors of a rotary table and a rotary shaft of an overall measurement light path connected upper rotary shaft type laser tracker, and belongs to the technical field of laser tracking measurement. The device comprises a microscope, a fluorescent microsphere, a substrate and a six-dimensional precision calibration displacement table, the microscope is located above the fluorescent microsphere, and the six-dimensional precision calibration displacement table is located above the substrate. The fluorescent microspheres are fixed on the substrate, and the substrate is fixed on the six-dimensional precise calibration displacement table. By combining the fluorescence microscope with the astigmatism imaging function, the precise displacement table, space coordinate calculation and other operations, the determination of the axis space linear equation of the upper and lower rotary tables of the whole measurement light path connected upper rotary shaft type laser tracker and the detection of the shaft radial runout in a complete rotation period are realized under one coordinate system; a necessary technical foundation is laid for realizing error compensation of the rotary table and the rotary shaft of the rotary shaft type laser tracker connected with an overall measurement light path and improving the tracking precision and the stability of the laser tracker.

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 test strips, in particular to a human saliva pepsin fluorescence immunochromatography test strip and a preparation method thereof.The human saliva pepsin fluorescence immunochromatography test strip is characterized in that a sample pad, a marking pad, a nitrocellulose membrane and a water absorption pad are pasted to the upper surface of a PVC plate, and the surface of the marking pad is sprayed with a pepsin marked antibody compound containing fluorescent microsphere marks; the fluorescence immunochromatography test strip provided by the invention can be used for specific detection of human saliva pepsin antigen, has a wide linear range, high sensitivity and high precision, can maintain the test value of the test strip to be basically unchanged for 14 days through accelerated destruction at 37 DEG C, shows good stability, and can be applied to detection of human saliva pepsin antigen. Meanwhile, the test strip is not interfered by amylase and lipase and has relatively high specificity, in addition, a method for detecting human saliva pepsin by adopting a fluorescence immunochromatography method has the advantages of operation detection, less time consumption, small matched equipment, low cost and the like, can meet the requirement of instant detection and has relatively high application value.

The invention belongs to the technical field of immunoassay rapid detection, and relates to a method for screening residues of traditional Chinese medicinal material aflatoxin B1 based on multicolor evolution fluorescence immunochromatography, which comprises the following steps: coupling red quantum dot fluorescent microspheres with an AFB1 antibody to prepare a detection probe; coupling or mixing the green quantum dot fluorescent microspheres with an AFB1 antigen to prepare an internal standard probe; respectively coating an internal standard probe and goat anti-mouse IgG on an NC membrane to complete preparation of a T line and a C line; preparing a test solution containing a detection probe, dripping the test solution on a test paper sample pad, and observing fluorescence colors of a T line and a C line by ultraviolet; and judging whether the residual quantity of the traditional Chinese medicine AFB1 exceeds the limit specification or not according to the T-line fluorescence color. By introducing the internal standard probe, a ratio fluorescence immunoassay system is constructed, so that the sensitivity and the accuracy of an analysis result can be improved, and the interference of a sample matrix is reduced; the used detection test paper is small in matrix influence and wide in application range, and can be used for rapid limited screening of AFB1 residues in traditional Chinese medicinal materials.

The invention discloses a kit for detecting Abeta1-40 protein as well as a preparation method and application of the kit. The invention provides a reagent for detecting Abeta1-40 protein. The reagent contains magnetic beads coupled with a capture antibody and / or a detection antibody labeled with fluorescent microspheres, the magnetic bead coupled with the capture antibody contains an A beta 1-40 protein capture antibody and a magnetic bead, and the diameter of the magnetic bead is 1.0-3.0 [mu] m; the detection antibody marked with the fluorescent microspheres contains an A beta 1-40 protein detection antibody and the fluorescent microspheres, and the diameter of the fluorescent microspheres is 190-400 nm. According to the present invention, the Abeta1-40 in the serum sample can be rapidly detected, the sensitivity is high, the repeatability is good, the sensitivity can achieve 0.12 pg / mL, and the data dispersion degree CV value can achieve 3.2%; the serum can be used as a sample for detection, the sample source is simple and easy to obtain, detection can be performed without living body puncture, and the use process is simple and convenient.

The invention discloses an immunodetection system and method based on a digital micro-fluidic chip, the system comprises a micro-fluidic chip, a heating unit, a magnetic field loading unit, a fluorescence detection unit and a control unit, the micro-fluidic chip comprises a liquid injection area, a magnetic mixing area, a magnetic separation area, a detection area, a waste liquid area and an electrode array; the electrode array covers each area and drives liquid drops in the areas to move; the control unit is used for controlling the electrode array to drive the movement sequence and position of the liquid drops in each region according to an immunodetection process; wherein the heating unit is used for regulating and controlling the immune reaction temperature of liquid drops in the magnetic mixing area, and driving the liquid drops to continuously move in the magnetic mixing area through the electrode array covered by the magnetic mixing area, so that samples, magnetic beads and fluorescent microspheres in the liquid drops are fully mixed and react. The immunoreaction time is greatly shortened, the magnetic mixing uniformity is greatly improved, and the immunodetection efficiency and accuracy are improved.

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 relates to the technical field of immunochromatography, in particular to a TSYl modified magnetic time-resolved fluorescent microsphere as well as a preparation method and application thereof, and the preparation method comprises the following steps: 1) preparing a surface aminated Fe3O4 magnetic core by a hydrothermal synthesis method; (2) carrying out double-bond modification on the surface of Fe3O4 through nucleophilic substitution reaction; 3) preparing surface hydroxyl modified polymer magnetic microspheres by a seed emulsion polymerization method; 4) through nucleophilic substitution reaction, performing tosyl modification on the surface of the magnetic microsphere; 5) preparing a time-resolved fluorescent dye Eu (III) complex; and 6) embedding a rare earth complex into the microspheres by a swelling method to prepare the surface tosyl modified magnetic time-resolved fluorescent microspheres. The TSYl modified magnetic time-resolved fluorescent microspheres prepared by the method have the advantages that the particle size range is 200-400nm, the particle size is uniform, and the monodispersity is good; the antibody can be directly combined with protein / antibody without activation; good magnetic responsiveness and fluorescence intensity are realized.

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 serum amyloid protein A immunochromatography detection reagent strip which comprises a reaction bottom plate, a sample pad, a combination pad, a chromatography membrane and absorbent paper, the chromatography membrane is arranged in the middle of the reaction bottom plate, the sample pad and the absorbent paper are arranged at the two ends of the reaction bottom plate respectively, the chromatography membrane is arranged in the middle of the reaction bottom plate, the combination pad is arranged between the chromatography membrane and the sample pad, the sample pad is partially overlapped on the combination pad, and the combination pad is partially overlapped on one side of the chromatography membrane. The absorbent paper is partially overlapped on the other side of the chromatography membrane; quantum dot fluorescent microspheres used for serum amyloid protein A antibody labeling and quantum dot fluorescent microspheres used for DNP-BSA labeling are solid-phase on the combination pad; the chromatographic membrane is provided with a detection line coated with a serum amyloid protein A detection antibody and a quality control line coated with a rabbit anti-DNP quality control antibody; and a detection line and a quality control line are sequentially arranged on the chromatography membrane in parallel at intervals along the chromatography direction. The invention further discloses a corresponding preparation method, a detection kit, a detection method and application.

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 invention provides a multicolor fluorescent microsphere, a tracer agent, a preparation method of the tracer agent and a detection method.The preparation method of the multicolor fluorescent microsphere comprises the steps that S1, a dispersion phase is provided, the dispersion phase comprises a hydrophilic polymer, a hydrophobic polymer, a hydrophilic fluorescent material, a hydrophobic fluorescent material and a non-polar solvent, and emission peaks of the fluorescent materials are different; providing a continuous phase, wherein the continuous phase comprises water and a surfactant; s2, respectively adding the dispersed phase and the continuous phase into an SPG film emulsifier, and extruding a film to form seed microspheres; removing the non-polar solvent to form a microsphere body; and S3, mixing the microsphere body solution, a monomer solution and an initiator, and carrying out polymerization reaction to form a shell layer coating the microsphere body, thereby obtaining the multicolor fluorescent microsphere.

The invention discloses a high-brightness quantum dot fluorescent microsphere and a preparation method thereof based on a base catalysis strategy. The preparation method comprises the following steps: (1) preparing a sulfydryl modified dendritic mesoporous silica microsphere; (2) assembling the sulfydryl modified dendritic mesoporous silica microspheres and quantum dots to obtain a dendritic silica / quantum dot assembly; (3) adding 3-mercaptopropyltriethoxysilane into the dendritic silicon dioxide / quantum dot assembly, and carrying out organosilane modification on the dendritic silicon dioxide / quantum dot assembly in a non-polar organic solvent by taking organic alkali as an alkali catalyst; and (4) growing a silicon dioxide shell layer on the surface of the organosilane modified dendritic silicon dioxide / quantum dot assembly by a # imgabs0 # method to obtain the high-brightness quantum dot fluorescent microsphere. According to the preparation method, fluorescence quenching of the oil-phase quantum dots in the process of coating silicon dioxide is avoided, and the excellent optical property of the oil-phase quantum dots is completely reserved.

The present invention relates to the field of microscopic imaging autofocus technology, specifically to a cell autofocus method and system for resisting interference from fluorescent microspheres in hydrogels. A set of hydrogel cell images containing fluorescent microspheres at different focal planes is collected, and for the set of hydrogel cell images containing fluorescent microspheres at different focal planes, the cell area weight and weighted gradient processing are calculated and stored in a preset clarity list; the clarity list is traversed, and the clarity scores of the hydrogel cell images containing fluorescent microspheres at different focal planes in the clarity list are performed to obtain the hydrogel cell image containing fluorescent microspheres with the highest clarity score and its corresponding Z-axis coordinate, and the Z-axis coordinate is used to control a preset motor to achieve autofocus. The present invention can adapt to various fluorescent microsphere densities without being affected by them, and has a wide range of applicability; the algorithm takes into account the presence of all cells in the field of view, is more holistic when focusing, and improves focusing accuracy.