63 results about "CELL DEBRIS" patented technology

The invention discloses an extraction preparation method and application of algae micromolecule PDRN, and relates to the technical field of biology. The invention establishes a process for extracting PDRN from microalgae, which comprises the following steps: carrying out cell disruption on microalgae liquid, and removing cell debris to obtain filtrate; the filtrate is subjected to DNA dissolution, DNA separation and DNA purification, and a PDRN solution is obtained; wherein the dry weight of the microalgae in the algae liquid is 0.1 to 0.333 g / mL. Through process optimization, the extraction efficiency and purity of the microalgae PDRN are improved, and the extracted PDRN has excellent anti-aging and anti-inflammatory effects.

The invention relates to the technical field of plant exosome-like extracts, in particular to a liquorice exosome-like extract as well as extraction and application thereof. The method comprises the following steps: firstly, removing impurities such as cells, cell debris, organelles, large vesicles and the like in homogenate by adopting a combination mode of low-speed centrifugation and medium-speed centrifugation, and further purifying the homogenate by adopting high-speed centrifugation after the low-speed centrifugation and the medium-speed centrifugation are completed, so as to obtain the exosome-like extract. And the setting of the increasing rotation speed is beneficial to finer separation of exosome-like particles with different sizes and densities, so that the purity and quality of the extract are improved. According to the method, all centrifugal operations are carried out in a low-temperature environment of 4 DEG C; the low-temperature condition is beneficial to maintaining the biological activity and stability of the sample, and the problems of protein denaturation, enzyme inactivation and the like caused by temperature rise are reduced, so that the quality and performance of the exosome-like extract are ensured.

The invention belongs to the technical field of biological pharmacy, and relates to a method for preparing tremella aurantialba polyphenol by utilizing tremella aurantialba TA58.21 fungus chassis cell fermentation, which comprises the following steps: adding a cooked soybean meal culture medium and a tremella aurantialba seed solution into a fermentation tank, introducing sterile oxygen, controlling the stirring rotating speed of the fermentation tank and the culture temperature to be 25-30 DEG C, continuously culturing for 5-8 days, and when the fermentation liquor becomes red, stopping the fermentation; when a large amount of foam is generated, closing an exhaust valve, keeping the tank pressure at 0.1 MPa, increasing the fermentation temperature to 50 DEG C, continuing to culture for 24 hours, increasing the stirring rotating speed of the fermentation tank, heating the fermentation liquor to 80-90 DEG C, keeping the temperature for 4-8 hours, stopping stirring, continuing to heat to 120 DEG C, and keeping the temperature for 30 minutes; centrifuging the fermentation liquor to remove precipitates and mycelia; collecting supernate, and pre-treating macromolecular substances and thallus fragments; collecting permeate liquid, and intercepting through a nanofiltration membrane to obtain a concentrated solution; and carrying out spray drying or freeze drying treatment on the concentrated solution to obtain a high-purity product. According to the invention, the tremella aurantialba polyphenol product with good water solubility, heat resistance, acid resistance and molecular weight of less than 10000 Da can be obtained.

The invention provides a construction method of a radioactive esophageal injury organ-like model. The construction method comprises the following steps: performing X-ray ionizing radiation on esophageal organs; wherein the esophageal organ comprises a rat esophageal organ, a mouse esophageal organ or a human esophageal organ; the radiation dosage is 0.5 to 4 Gy, and the radiation dosage rate is 1.5 to 2 Gy / min. Experimental results show that along with increase of irradiation dose, connection around spheres of human esophageal organs, mouse esophageal organs and rat esophageal organs is loose, cell debris is increased, and the sizes are smaller than those of normal organs at the same time. The construction of the radioactive esophageal injury organ-like model is of great significance for deeply understanding the injury mechanism of ionizing radiation to the esophagus, searching for effective protection and treatment measures and promoting the development of personalized radiotherapy.

The invention relates to the technical field of medical detection, in particular to a microneedle array-containing S100A9 sample sampling and detecting device which comprises a base, a limiting column and a color developing assembly, the limiting column is fixedly connected to the top end of the base, and a working sliding groove for inserting the color developing assembly is formed between the limiting column and the base; a sampling groove is formed in the top end of the base, the inner bottom wall of the sampling groove is fixedly connected with a microneedle assembly used for puncturing the skin of a patient for sampling, and the top of the microneedle assembly communicates with a pretreatment assembly used for filtering an extracted sample; driving force is provided for the pretreatment assembly in the process that the color developing assembly is inserted into the working sliding groove, and the filtered sample is input into the color developing assembly to be detected. According to the invention, convenient sampling is realized through the microneedle assembly, and a sample is pertinently filtered by using the pretreatment assembly, so that macromolecular interference impurities such as cell debris and hemoglobin are effectively removed, the interference on S100A9 detection is reduced, and the accuracy and reliability of S100A9 detection are improved.

The invention discloses a sleep-promoting nasal spray based on mesenchymal stem cell-derived exosomes and preparation and application thereof.The preparation method comprises the steps that 1, mesenchymal stem cells separated from umbilical cord tissue are cultured to P3-P5 generations, cell culture supernate is collected, cells are digested, and then cell debris and macromolecular impure protein are removed through pretreatment; (2) filtering and concentrating the pretreated cell culture supernatant and replacing a part of solution; (3) carrying out degerming filtration on the solution obtained in the step (2) through a 0.22 [mu] m filter membrane to obtain an MSC-Exos solution; and (4) adding a stabilizer with a proper concentration into the MSC-Exos solution to obtain a compound MSC-Exos solution, namely the sleep-promoting nasal spray. The product is a cell-derived natural active substance, can significantly improve sleep, and does not involve any pharmaceutical ingredient.

The invention discloses a purification method of cord blood mesenchymal stem cell source exosome, and relates to the technical field of bioengineering. The method comprises the following steps: firstly, carrying out pH regulation pretreatment on cell culture supernate through a technical path of combining gradient centrifugation and membrane filtration, and sequentially removing cell debris and large-particle impurities by adopting multi-stage differential centrifugation; then carrying out selective protein precipitation by utilizing polyethylene glycol with specific molecular weight, and realizing fine separation of the exosome by combining sucrose-meglumine diatrizoate density gradient centrifugation; and finally, terminal purification is completed under low-temperature and low-pressure conditions through a polyethersulfone membrane filtration system with an optimized pore size. According to the method, centrifugal parameters, reagent concentration and filtering conditions are precisely regulated and controlled, so that the particle size of the obtained exosome is intensively distributed in the range of 30-150nm, the protein purity reaches 95% or above, the recovery rate exceeds 85%, and the separation efficiency and quality of the exosome are remarkably improved.

The invention belongs to the technical field of cancer immunotherapy, and particularly relates to iterative radiation-resistant tumor cell debris, a preparation method thereof and an application thereof in tumor resistance. The anti-radiation tumor cells generated by repeated radiation exposure show higher TMB (Tetramethylbenzidine) and induce stronger immunoreaction, so that a personalized vaccine is developed based on the method, and the frozen fragment vaccine is obtained by freeze thawing and inactivation of the radiation-resistant tumor cells. The RR-FDV exhibits enhanced immunogenicity as compared to a conventional radiosensitive counterpart. In the present disclosure, RR-FDV exhibits superior activation of antigen presenting cells, while in vivo, it recruits more dendritic cells to the vaccination site, promotes DC-mediated antigen presentation, and induces stronger antigen-specific T cell toxicity.

The present invention relates to a method for producing and purifying a viral vector (V) wherein the viral vector (V) is produced from cells in a fluid (F) and wherein the fluid (F) comprising the viral vector (V) is conducted through a tangential flow filtration device (2) for purification, whereby the viral vector (V) is purified in the fluid (F). It is proposed that the cells and cell debris from the cells are at least partially separated from the viral vector (V) by a fluidized bed centrifuge (5) before purification by the tangential flow filtration device (2), which cells and cell debris are contained in a fluid (F) containing the viral vector (V) after production of the viral vector (V).

The utility model discloses enzymatic hydrolysate centrifugal separation and purification equipment for active peptide powder processing, and relates to the technical field of active peptide powder processing, the enzymatic hydrolysate centrifugal separation and purification equipment comprises a centrifugal machine, a tank body, a finished product tank, a liquid outlet pipe, a connecting pipe and a filter box, the lower part of the centrifugal machine is communicated with the finished product tank, the tank body is arranged at one side of the finished product tank, and the liquid outlet pipe is arranged at the other side of the finished product tank; the centrifugal machine is communicated with the finished product tank through a pipe body, and the centrifugal machine is communicated with the finished product tank through the liquid outlet pipe. According to the utility model, enzymatic hydrolysate is primarily purified through the centrifugal machine, solid particles (such as cell debris and unreacted substrates) in the enzymatic hydrolysate are separated from liquid, and the separated finished product liquid directly enters the filter box; the finished product liquid sequentially passes through the arc-shaped filter screen, the filter plate I and the filter plate II to perform membrane filtration on small molecular impurities and part of macromolecular substances, and the centrifuged finished product liquid is refined, so that centrifugation and membrane filtration are performed simultaneously, and the production efficiency is improved.

Process for simultaneous production of sugars and lipids from a fermentation broth comprising the following steps: (a) feeding at least one microorganism, preferably an oleaginous microorganism, to a fermentation device obtaining a fermentation broth comprising an aqueous suspension of cellular biomass comprising sugars and lipids; (b) optionally, at the end of the fermentation, subjecting the fermentation broth obtained in said step (a) to concentration obtaining a fermentation broth comprising an aqueous suspension of concentrated cellular biomass comprising sugars and lipids; (c) subjecting the fermentation broth obtained in said step (a) or the fermentation broth comprising an aqueous suspension of concentrated cellular biomass comprising sugars and lipids obtained in said step (b) to hydrolysis in the presence of at least one organic acid having from 1 to 6 carbon atoms, preferably from 1 to 3 carbon atoms, said hydrolysis being carried out at a pH comprised between 0.6 and 1.6, preferably comprised between 0.9 and 1.3, obtaining a reaction mixture comprising cellular debris, an aqueous phase and an oily phase; (d) subjecting the reaction mixture obtained in said step (c) to separation obtaining a solid residue comprising cellular debris, an aqueous phase comprising sugars and an oily phase comprising lipids. The sugars obtained from the aforesaid process can be advantageously used, for example, as carbon sources in fermentation processes for the production of alcohols (e.g., ethanol, butanol), lipids, diols (e.g., 1,3-propanediol, 1,3-butanediol, 1,4-butanediol, 2,3-butanediol), or in chemical synthesis processes for the production of other intermediates or chemical products. Said alcohols and lipids can in turn be advantageously used in the production of biofuels (e.g., biodiesel or 30 "Green Diesel"), which can be used as such or in a mixtures with other automotive 33 fuels, while said diols can be used in the production of products such as, for example, bio-butadiene, which can in turn be used in the production of rubbers (e.g., polybutadiene or its copolymers). The lipids obtained from the aforesaid process can be advantageously used, for instance, for the production of biofuels that can be used in diesel or aviation engines, or these lipids can be subjected to metathesis processes in order to obtain biofuel precursors, waxes, plastics, cosmetics, and personal care articles. The aforesaid process is particularly useful in the case of a biorefinery.

A method for producing a target product from biomass waste, comprising the following steps: (A) putting bacteria A into a first culture medium made from biomass waste and performing degrading fermenta

The invention discloses a tumor cell detection kit, and relates to the technical field of detection kits, the tumor cell detection kit comprises a detection uniformizing module, the detection uniformizing module comprises: a first gear rotatably arranged around a first axis of the first gear; the driving disc is in coaxial running fit with the first gear; the gear assembly further comprises at least one set of strip-shaped holes and arc-shaped holes which are matched with each other, the strip-shaped holes are formed in the radial direction of the first gear, the strip-shaped holes are in guide fit with movable assemblies, the movable assemblies comprise matching shafts matched with the arc-shaped side faces of the arc-shaped holes, and the strip-shaped holes and the arc-shaped holes are formed in the first gear and the driving disc respectively. According to the uniform detection module, through the matching mode of the driving disc and the first gear, the overall structural stability is high, the working requirement of low-speed centrifugation can be met, the working requirement of cell debris separation can be met, and the working efficiency is improved. The adaptability of the detection kit is improved.

The present invention relates to the field of biomedical technologies and provides a method for efficiently capturing neutrophils from peripheral blood for single-cell sequencing by lysing red blood cells. The method includes the following steps: collecting human peripheral blood into an anticoagulant tube containing ethylenediaminetetraacetic acid; taking the ethylenediaminetetraacetic acid anticoagulated whole blood, adding a red blood cell lysate, lysing at room temperature, waiting until the turbid mixed blood solution becomes clear and transparent, waiting for 1 minute, then adding a FACS cell resuspension solution to terminate red blood cell lysis, and centrifuging; adding the FACS cell resuspension solution to slowly resuspend the cells, and centrifuging; adding the FACS cell resuspension solution to slowly resuspend the cells, and centrifuging; adding a phosphate buffer solution to slowly resuspend the cells, and centrifuging. By selecting a suitable anticoagulant, precisely controlling the red blood cell lysis conditions, and performing meticulous cell resuspension and centrifugation operations, the present invention can ensure high neutrophil activity and a low cell debris rate, provide a suitable single-cell suspension for single-cell RNA sequencing, and help analyze the characteristics and functions of various neutrophil subsets.

The invention relates to a battery monomer, a battery and a power utilization device. Each single battery comprises a battery cell and a first insulating part, and the first insulating part wraps the battery cell; the first insulating part is provided with an inner side surface facing the battery cell and an outer side surface deviating from the battery cell, and a first through hole is formed in the first insulating part; the first through hole penetrates through the inner side surface and the outer side surface, and the cross-sectional area of the first through hole on the inner side surface is smaller than that of the first through hole on the outer side surface; alternatively, the first through hole comprises a plurality of first through holes, and each first through hole penetrates through the inner side face and the outer side face. According to the battery monomer, the flow area of the electrolyte can be increased, the flowability of the electrolyte is better, electrolyte infiltration is facilitated, meanwhile, cell falling objects or other foreign matters can be blocked, the condition of metal filing falling or carbon powder falling is improved, the safety is improved, and the service life is prolonged.

The invention discloses a stem cell-based microvesicle as well as a preparation method and application thereof, belongs to the technical field of biological tissue engineering, and particularly relates to a preparation method of the stem cell-based microvesicle, which comprises the following steps: culturing stem cells until the cell fusion degree is 80-90%, performing hypoxia treatment, and cleaning for 2-3 times; standing and culturing the cleaned stem cells, then adding a calcium ion carrier, and carrying out ultrasonic treatment; the serum-free basic culture medium containing the calcium ion carrier is immediately absorbed and removed, the cells are cleaned through PBS, culture is conducted for 24 h, and cell supernatant is collected and merged respectively; performing first centrifugation to remove cell debris; performing second centrifugation to remove dead cells and large cell debris; carrying out third centrifugation to remove organelles; fourth centrifugation is carried out, large vesicles are removed, and a final supernatant is obtained; and filtering and centrifuging to obtain a microvesicle precipitate. The preparation method of the microvesicles aims at systematically improving the yield and biological activity of the stem cell microvesicles through low-oxygen pre-stimulation, ultrasonic-calcium ion carrier synergistic induction and differential centrifugation.

The utility model discloses a stem cell conditioned medium filter device which comprises a filter device shell, a filter tank is arranged in the filter device shell, a square sleeve is arranged on the lower side in the filter tank, a coarse-mesh filter screen is fixedly arranged in the square sleeve, and a filter screen is arranged in the coarse-mesh filter screen. A plurality of fine-hole filter screens fixed in the square sleeve are fixedly arranged below the coarse-hole filter screen; a sealing plate fixed in the square sleeve is arranged above the coarse-mesh filter screen, and a first stepping motor is fixedly mounted in the sealing plate. According to the utility model, the situation that the filter holes in the filter screen are blocked by cell debris, dead cells or metabolic wastes in the stem cell conditioned culture medium and even penetrate through the filter holes due to relatively intense impact generated between the filter screen and the stem cell conditioned culture medium when the filter screen is used for filtering the stem cell conditioned culture medium is prevented; through the technical scheme, the filtering effect of the equipment is improved.

The application relates to a method for purifying umbilical cord blood mesenchymal stem cell-derived exosomes, and relates to the technical field of bioengineering. The method combines gradient centrifugation and membrane filtration. First, the cell culture supernatant is pretreated by adjusting the pH, and cell debris and large-particle impurities are removed by multi-stage differential centrifugation. Then, selective protein precipitation is carried out by using polyethylene glycol with a specific molecular weight, and the exosomes are finely separated by sucrose-meglumine diatrizoate density gradient centrifugation. Finally, terminal purification is completed under low-temperature and low-pressure conditions by using a polyether sulfone membrane filtration system with optimized pore size. By precisely controlling the centrifugation parameters, reagent concentration and filtration conditions, the exosomes obtained by the method have a particle size concentrated in the range of 30-150 nm, a protein purity of more than 95%, and a recovery rate of more than 85%, and the separation efficiency and quality of the exosomes are significantly improved.

The invention discloses a method for identifying chronic granulocyte immune typing by using a flow cytometer. The method comprises the following steps: incubating and purifying a to-be-detected sample and a fluorescein-labeled antibody composition; the method comprises the following steps: carrying out flow cytometry detection on resuspended cells: firstly, circling a liquid flow stable interval by utilizing Time / CD45, FSC INT / PEAK and FSC INT / SSC INT scatter diagrams, and removing sticky bodies and cell debris; all leukocyte groups are circled through combination of CD45 and SSC, and mononuclear cells are circled in all leukocyte gates through CD45 / SSC gate setting; excluding CD 19 + B lymphocytes; cD7 + T and NK cells are excluded; cD2 + T and NK cells are eliminated; immature and mature granulocytes of CD 16 and CD24 < + > are excluded through CD 16 / CD24 gate setting; the method comprises the following steps of: screening CD14-CD16, removing basophilic granulocytes of CD14-CD16-and residual NK (Natural Killer) cells, dividing a cross gate to determine the proportion of CD 14 + CD16-cMo cells, CD 14 + CD16 + iMo cells and CD 16 + CD 14 ncMo cells, and determining the proportion of the CD14 + CD16-cMo cells, the CD14 + CD16 + iMo cells and the CD16 + CD14 ncMo cells; cD34 / SSC gate setting is carried out, and CD34 + original cells are circled to calculate the proportion.

This invention provides an in vivo circulating tumor cell (CTC) clearance system, relating to the field of medical device technology. The in vivo CTC clearance system provided in this invention utilizes switching between three pathways: a CTC separation pathway, a CTC electroporation pathway, and a cell debris removal pathway. Based on a metal filter membrane, it continuously separates CTCs in vivo and periodically performs electroporation to break down and remove the collected CTCs. This avoids clogging of the filter pores due to continuous metal filter operation, allowing for continuous, periodic operation to clear all existing and newly generated circulating tumor cells (CTCs) in the body.

The invention relates to a method for operating a biological treatment plant (1) comprising a purification device (2) for removing cell debris from a cell culture fluid (10), in which the purification device (2) comprises a fluidized bed centrifuge (8) having at least one centrifugal chamber (11), in which the cell culture fluid (10) is separated into at least a waste fraction (12) and an output fraction (13), wherein the separation of the cell culture fluid (10) into a waste fraction (12) and an output fraction (13) comprises: a loading step (14) during which the centrifugal chamber (11) is loaded with cells of the cell culture fluid (10) up to a capacity lower than or equal to the maximum cell loading capacity of the centrifugal chamber (11); an overloading step (15) during which the centrifugation chamber (11) is further loaded with a cell culture fluid (10), whereby cells are flushed out of the centrifugation chamber (11); collecting the cells flushed out during the overload step (15) as the waste fraction (12); and collecting cells from the chamber as the output fraction (13).

The invention discloses a preparation method for prolonging the in-vivo half-life period of an artificial exosome. The preparation method comprises the following steps: S1, extracting the exosome; s1.1, cell culture: culturing the human liver Kupffer cells for 7 days; s1.2, primary centrifugation: collecting a culture solution, and centrifuging to remove cells and large fragments; s1.3, secondary clarification: transferring the supernatant to a new tube, and centrifuging to further remove smaller cell debris; s1.4, ultracentrifugal precipitation of the exosome: transferring the supernate again, and centrifugally precipitating the exosome; s1.5, re-suspending the exosome: gently re-suspending the precipitate with PBS, and carrying out ultracentrifugation to remove impurities; s2, platelet membrane component extraction; s3, combining the exosome with the platelet membrane component. According to the invention, the in-vivo half-life period of the exosome is greatly prolonged by using a simpler process and cost, and the half-life period can reach 24-48 hours.

The invention provides a bioartificial liver supporting system which at least comprises a blood input branch pipeline, a plasma separation branch pipeline, a biological purification branch pipeline and a plasma return branch pipeline which are connected in sequence, and a liquid separation bottle is arranged between the biological purification branch pipeline and the plasma return branch pipeline; the liquid separation bottle comprises a bottle body, at least three partition plates are arranged in the bottle body, the heights of the partition plates are sequentially reduced in the circumferential direction so that the interior of the bottle body can be at least divided into three chambers with the water level heights sequentially reduced in the circumferential direction, the biological purification branch pipeline is communicated with the chamber with the highest water level height, and the slurry return branch pipeline is communicated with the chamber with the lowest water level height. According to the bioartificial liver supporting system, the liquid separation bottle is arranged, and the water level height difference of the multiple chambers formed in the liquid separation bottle is utilized, so that seed cells and cell debris contained in blood plasma are deposited, the situation that the seed cells and the cell debris are infused back into the body of a patient along with the blood plasma to cause discomfort of the patient is avoided, and the use safety of the bioartificial liver supporting system is improved.

The invention provides an engineered small extracellular vesicle as well as a preparation method and application thereof. The preparation method comprises the following steps: S1, removing cell debris in a sample to obtain supernate; extracting from the supernate to obtain the small extracellular vesicles; s2, preparing a phospholipid solution; dissolving the polypeptide A with Tris. HCl, and then adding the phospholipid solution for reaction to obtain a coupled polypeptide A system; taking EDC and sulfo-NHS, then adding rhodamine polypeptide B and the small extracellular vesicles, and dissolving the rhodamine polypeptide B and the small extracellular vesicles in tris (hydroxymethyl) aminomethane hydrochloride for reaction to obtain a coupled rhodamine polypeptide B system; and S3, uniformly mixing the coupled FITC polypeptide A system and the coupled rhodamine polypeptide B system, and performing dialysis to obtain the engineered small extracellular vesicles. The obtained engineered small extracellular vesicles are stable, can be supplied continuously, can reduce neurotoxicity effect and relieve the condition of Parkinson's disease, and provide a new target and a new way for clinical treatment of Parkinson's disease.

The invention provides an acellular matrix for treating stress urinary incontinence as well as a preparation method and application of the acellular matrix. Splitting the soft tissue by using a cell lysis solution to remove cell debris; removing nucleic acid, degreasing and then micronizing; and adding the soft tissue particles and the soft tissue cross-linked particles into a phosphate buffer solution according to a ratio, mixing and emulsifying to obtain the acellular matrix for injection for treating stress urinary incontinence. The cell removal rate reaches 99%, the fat residue is less than 0.3%, and the injection is injected around the sphincter of the urethra (middle section or bladder neck) and / or in the urethra according to 0.5 ml / point and 3-4 points during clinical application to play roles in filling, connecting and repairing tissues, so that the damaged urethra supporting structure is repaired, the structural integrity and strength of connective tissues and the sphincter around the urethra are maintained, and the application prospect is wide. Therefore, the symptom of stress urinary incontinence is improved.

A particulate lyophilized platelet lysate composition suitable for use as a cell culture medium can include growth factors, cytokines, and chemokines released from lysis of source platelets, wherein cellular debris from the source platelets is removed (partially or fully) by filtration. The growth factors, cytokines, and chemokines are lyophilized to form a particulate lyophilized platelet lysate composition.

This utility model discloses a vaccine microcarrier recovery filter with a clean structure, including a workbench. A cleaning tank is located at the top of the workbench, and a first water tap is located at one end of the cleaning tank. A water storage tank is located at the bottom of the workbench. A filter groove is formed on the inner wall of the workbench, and a filter is movably installed in the filter groove. This utility model achieves automatic rotational cleaning of the filter through the cooperation of a linear motor and a waterproof motor. This dynamic cleaning method is more efficient than static cleaning, and can more thoroughly remove cell debris, culture medium residues, and microbial contamination from the filter surface, ensuring the cleanliness and effectiveness of the filter.

The invention discloses an LSPR (Local Surface Plasmon Resonance) micro-fluidic chip system for rapid typing detection of various virus antigens, and particularly relates to the cross technical field of biomedical engineering, a micro-nano sensing technology, molecular diagnosis and micro-fluidic chip application. According to the present invention, by introducing the PEGylation anti-fouling layer and the BSA sealing treatment, the non-specific adsorption interference is effectively reduced, and compared with the problems that the traditional antigen rapid detection (Ag-RDT) is easily influenced by the virus epitope mutation and the variation is difficult to distinguish, the system can utilize the LSPR peak position red shift difference caused by the combination of different variation antigens and the specific antibody to achieve the precise typing of the multi-variant virus, meanwhile, white light dark field hyperspectral detection is combined with baseline correction, noise filtering and other algorithms, spectral signals can be accurately extracted, and even if interference components such as impurity protein and cell debris exist in a sample, feature signals of a target antigen can still be stably recognized.

The invention belongs to the technical field of biological medicine and clinical medicine, and particularly relates to an efficient purification method of brain tissue small extracellular vesicles. The purification method comprises the following steps: cutting mouse brain tissues into uniform small blocks by using a steel wire mesh, digesting and dissociating, filtering, taking filtrate, and fixing the volume of the filtrate by using an EBSS buffer solution to obtain a tissue supernatant; the method comprises the following steps: sequentially removing complete cells, cell debris, plasma membranes and large vesicles through gradient centrifugation, filtering, taking filtrate, carrying out ultracentrifugation to obtain sEVs precipitates, and resuspending the sEVs precipitates with a PBS buffer solution to obtain a sEVs crude purification product; and preparing a single sucrose pad from a sucrose solution with the concentration of 25-35% and a PBS buffer solution, and carrying out ultracentrifugation on the sEVs crude purification product, wherein the obtained precipitate is the purified brain tissue sEVs. The purification method provided by the invention solves the problems of tedious flow, long consumed time, low yield, poor purity and the like in the existing sEVs purification technology.

The invention discloses an integrated micro-fluidic metasurface chip for real-time detection of exosome (POCT). The chip is formed by bonding a micro-fluidic structure layer and an optical sensing substrate layer, and sequentially comprises a sample / buffer solution converging region, a gradient film pretreatment region, a snake-shaped mixing region, a micro-column enrichment region and a multi-channel super-surface detection region in the flow direction. The coarse filtration membrane and the fine filtration membrane realize graded removal of cell debris and platelets, and are matched with a back-flushing structure to reduce blockage of the filter membranes; the microcolumn array is modified by a general exosome antibody and is used for efficient immune enrichment and mild washing or elution of exosome; the multi-channel metasurface array is used for respectively fixing different disease-related probes and carrying out high-sensitivity parallel optical detection on multiple markers of exosomes. According to the present invention, the single-chip integration of exosome pretreatment, enrichment and detection is achieved, the sample loss is reduced, the detection sensitivity and the flux are improved, and the method is suitable for the POCT application scenarios such as tumor early screening. Figure 1 is a structural diagram of the integrated micro-fluidic metasurface sensing chip provided by the invention.