36 results about "Tissue homogenate" patented technology

The invention belongs to the field of immunological detection, and provides a double-antibody sandwich ELISA kit for detecting the content of human beta-NGF. The enzyme-linked immunosorbent assay kit constructed by the invention is based on a double-antibody sandwich method, and is combined with a biotin-streptavidin signal amplification system and an enzyme developing amplification system, so that the detection sensitivity and specificity are greatly improved, and low-content samples can be accurately and quickly detected. The kit shows excellent stability and specificity and can be used for quantitatively detecting the content of beta-NGF in samples such as human serum, cell supernatant or tissue homogenate, so that an objective detection basis is provided for diagnosis, curative effect evaluation and pathogenesis research of related diseases.

The invention provides a preparation method of a terminal-peptide-free bovine type I collagen product. The preparation method comprises the following steps: obtaining a crushed bovine tissue raw material without impurities; the preparation method comprises the following steps: soaking a cattle tissue raw material in acid, and homogenizing to obtain cattle tissue homogenate; protease is added into the bovine tissue homogenate in batches for enzymolysis, the enzymolysis temperature is controlled to be 2-6 DEG C, after salting-out precipitation, pH value adjustment and redissolution are carried out on the bovine type I collagen extracting solution obtained in the middle batch, protease is added again for enzymolysis, salting-out precipitation, pH value adjustment and redissolution are carried out again, and the bovine type I collagen extracting solution is obtained. Repeating for multiple times to obtain a terminal-peptide-free bovine type I collagen solution; adjusting the pH value of the terminal-peptide-free bovine type I collagen solution, salting out, precipitating, centrifuging and washing to obtain terminal-peptide-free bovine type I collagen precipitate; and purifying the terminal-peptide-free bovine type I collagen precipitate to obtain the terminal-peptide-free bovine type I collagen product. According to the telopeptide-free bovine type I collagen product provided by the invention, no residual telopeptide is detected, and the purity is high.

The invention is applicable to the field of food processing, and provides a preparation method of pig brain oligopeptide capable of promoting learning ability, which comprises the following steps: selecting qualified pig brain subjected to inspection and quarantine, adding clear water, stirring to prepare homogenate, heating to 90-95 DEG C, and keeping for 15 minutes; cooling the obtained tissue homogenate, and carrying out first enzymolysis; adjusting the pH value of the first enzymolysis system, standing and precipitating, separating supernate and precipitate, and adding clear water into the precipitate to re-homogenate; carrying out secondary enzymolysis on the precipitate homogenate; adjusting the pH value of the second enzymolysis system, standing and precipitating, and separating supernate and precipitate; and combining the supernatant obtained by the first enzymolysis with the supernatant obtained by the second enzymolysis, concentrating, dialyzing and drying to obtain the pig brain oligopeptide. The method is simple and convenient to operate, low in cost and high in raw material utilization rate, and the obtained product is high in purity and easy to absorb, has the physiological functions of promoting learning and memory, tonifying and strengthening the brain and the like, and has wide industrial application prospects.

The invention discloses a tissue homogenizer for bulb vegetable detection, and relates to the technical field of food detection.The tissue homogenizer comprises a machine base and a limiting ring, the top end of the machine base is fixedly connected with the limiting ring, a lower cylinder is placed on the inner side of the limiting ring, the top end of the lower cylinder is spirally connected with an upper cylinder, and the top end of the upper cylinder is spirally connected with an end cover; a connecting shaft is rotatably connected to the inner side of the end cover, a source restraining mechanism and a defoaming mechanism are arranged on the outer side of the connecting shaft, the source restraining mechanism is arranged to forcibly press surface slurry (the area close to the liquid level) to the bottom of the cavity, and the problems that a traditional cutter swings the slurry outwards and impacts the liquid level are solved through the design; cutting off the path of air involved into the slurry from the source; and the slurry smoothly and downwards flows along the inclined surface of the cutter without rolling and air entrapment phenomena, so that bubbles generated by turbulent flow are eliminated from the source.

1. Name of the product of this design: Automated extraction equipment for purification and concentration of exosomes. 2. Purpose of the product of this design: This device is used to extract, purify and concentrate exosomes from cell culture supernatant and tissue homogenate. 3. The key point of the design of this product lies in its shape. 4. The picture or photo that best illustrates the design points: three-dimensional picture.

The invention discloses a method for extracting cell nucleuses from animal tissues, and relates to the technical field of biology. The background technology indicates that cell nucleus extraction is crucial to multidisciplinary research, but a traditional method has the problems of complex operation, time consumption, low purity, easiness in damaging cell nucleuses and the like, and the prior art is still limited although the prior art is improved. The method comprises the following steps: preparing a sample, and taking fresh or cryopreserved animal tissues; homogenizing the tissues, namely putting the tissues into a homogenizer, and homogenizing at a low speed by adding a cracking buffer solution; carrying out cell lysis, and incubating homogenate on ice; centrifugally separating, centrifuging at low temperature and medium speed, taking precipitate, and discarding supernatant; core washing, re-suspending the precipitate in a washing buffer solution, uniformly mixing and filtering; performing core extraction, re-suspending the precipitate in a suspension buffer solution, and lightly and uniformly mixing; purifying and washing, carrying out density gradient centrifugation on the suspension, dyeing with trypan blue, taking a core layer, and washing for subsequent experiments. The nuclear extraction lysis buffer solution has two proportions of LB1 and LB2, different tissues are suitable for different proportions, and an enzyme inhibitor can be added as required. The method can be used for rapidly extracting various animal tissue cell nucleuses, is complete in shape, large in quantity and few in impurities, is suitable for different tissues, and can be applied to downstream experiments such as single cell nucleus sequencing.

The invention belongs to the technical field of disease control in aquaculture, and particularly relates to a hippocampus enteritis pathogen separation and identification and medication screening method based on a high-throughput sequencing technology, which comprises the following steps: collecting intestinal tissues of a diseased hippocampus and a healthy hippocampus, respectively preparing tissue homogenate, and freezing for preservation; the method comprises the following steps: extracting DNA (Deoxyribose Nucleic Acid) of microorganisms of an intestinal tissue sample, performing 16SrDNA high-throughput sequencing, analyzing intestinal flora composition, and determining dominant bacteria; selecting and screening a culture medium according to dominant bacteria, and performing pathogenic bacterium separation and purification on intestinal tissues of the diseased hippocampus; dNA of the purified strain is extracted, 16S rDNA amplification and sequencing are carried out, and the species of pathogenic bacteria are identified through sequence comparison; according to the method, the diversity and abundance differences of microorganisms in the sick hippocampus and the healthy hippocampus are compared through a high-throughput sequencing technology, and targeted separation, identification and drug sensitivity screening are further carried out by taking statistically significant difference bacteria as the guidance of potential pathogenic bacteria.

This utility model discloses a biological tissue homogenizer, specifically relating to the field of cell tissue homogenization technology. It includes a homogenization mechanism comprising a base and a support frame. A placement mechanism is slidably connected above the support frame, and a hydraulic cylinder is mounted above the support frame. This utility model achieves automatic cleaning and surface moisture adsorption of the homogenizing rod through the coordinated operation of the placement mechanism, cleaning mechanism, wiping mechanism, connecting rope, and water tank. The placement mechanism and cleaning mechanism are linked by guide wheels and slide rails, allowing for simultaneous placement and removal of the homogenizing tube, reset of the cleaning mechanism, and preparation for cleaning the homogenizing rod with a single pull, simplifying the operation process. The wiping components (door panel, absorbent cotton) employ an automatic reset design with torsion springs to ensure unobstructed movement of the homogenizing rod, and the absorbent cotton is periodically squeezed by a squeezing block to expel moisture, extending its service life.

The invention belongs to the field of animal models, particularly relates to a granulomatous mastitis animal model and a construction method thereof, and aims to solve the problems of low pathological matching degree and poor repeatability of an existing model. The construction method comprises the following steps: selecting an SPF-grade BALB / c female mouse (10 weeks old, 20-22 g), preparing human mammary gland fresh lesion tissue homogenate and lesion origin staphylococcus aureus liquid (1.4 * 10 < 8 >-1.5 * 10 < 8 > CFU / mL), mixing and emulsifying with a Freund's incomplete adjuvant according to a ratio of 1: 1: 1, injecting to the third pair of mammary glands of the mouse, and completing modeling in two weeks. The pathological characteristics of the model are highly matched with human diseases, the success rate is high, the repeatability is strong, and the model can be used for GM pathogenesis research and therapeutic drug screening.

The utility model provides a food tissue homogenizer, and relates to the technical field of food tissue homogenizers, the food tissue homogenizer comprises a homogenizer base, a mounting frame and a homogenizing cup, through the arrangement of a cleaning device, when cleaning and maintenance are needed, a limiting rod can be aligned with a rectangular groove, and the homogenizing cup is arranged in the rectangular groove; meanwhile, one end of the mounting ring block is aligned with the inner wall of the mounting frame to clamp the mounting ring block and the mounting frame at the same time, then the spiral and the homogenizing cup are fixed, and similarly, the crushing cutter is driven to rotate in the same way as the crushing and homogenizing treatment process; the first brushes and the second brushes can be driven to rotate in the inner wall of the mounting frame and the inner wall of the homogenizer to clean the inner walls of the mounting frame and the homogenizer, the two second brushes are arranged on the two sides of the crushing cutter, dead corners between blades of the crushing cutter and the mounting frame can be cleaned, and therefore the hands of cleaning personnel can be prevented from being accidentally injured, and the cleaning efficiency is improved. The problem of potential safety hazards during cleaning and maintenance is reduced, the homogenizer is convenient to clean, and the use efficiency of the homogenizer is improved.

The invention belongs to the technical field of biological medicine, and particularly relates to a kit for breast cancer diagnosis and application thereof. The kit comprises an elisa plate coated with a captured antibody and an HRP labeled antibody working solution, the capture antibody is a monoclonal antibody Y2A-1; the HRP labeled antibody is a monoclonal antibody P4X-2; the heavy chain amino acid sequence of the monoclonal antibody Y2A-1 is as shown in SEQ ID NO.1, and the light chain amino acid sequence of the monoclonal antibody Y2A-1 is as shown in SEQ ID NO.2; the heavy chain amino acid sequence of the monoclonal antibody P4X-2 is as shown in SEQ ID NO.3, and the light chain amino acid sequence of the monoclonal antibody P4X-2 is as shown in SEQ ID NO.4. The kit for breast cancer diagnosis can quantitatively detect the PPIP5K2 protein content in a human tissue homogenate sample, serves as a breast cancer auxiliary diagnosis means for highly expressing PPIP5K2 proteins, and has a wide application prospect.

This utility model discloses a tissue filtration device, including a centrifuge tube, a first filter element, a second filter element, and a propulsion component. The centrifuge tube has an inner cavity with an opening. The first filter element is installed in the inner cavity and has a first filter chamber. The first filter chamber has a first filter hole and an opening, which are connected to the first filter chamber. Tissue homogenate can pass through the first filter hole, while connective tissue is blocked by the first filter element and remains in the first filter chamber. The second filter element is installed at the bottom of the first filter chamber and has a second filter chamber. The second filter chamber has a second filter hole and an opening, the diameter of which is smaller than that of the first filter hole. The second filter hole and the opening are connected to the second filter chamber, where connective tissue is collected. The propulsion component is located in the first filter chamber and is used to propel the tissue homogenate towards the second filter element. The propulsion component applies pressure to the tissue homogenate. This utility model can reduce the residual rate of connective tissue in tissue homogenate, shorten processing time, and improve filtration efficiency.

This invention discloses an in-situ vitrification-semi-solid nucleic acid room-temperature stable preservation solution, its preservation method, and a reagent kit, belonging to the field of biological sample preservation technology. The preservation solution comprises a double buffer system, a mild chelation system, a vitrification framework, a thermoresponsive gelling system, an oxidative protection system, a low-toxicity antibacterial system, and water. The thermoresponsive gelling system forms a weakly reversible gel network within the range of 18-37°C after the sample is mixed with the preservation solution, placing the sample nucleic acid in a confined microenvironment with low diffusion and low water activity. This invention achieves long-term stable preservation of nucleic acid samples at room temperature through the synergistic effect of a triple mechanism of "low water activity - thermoresponsive gelling - multi-target passivation." This invention eliminates the need for cold chain transportation, is simple to operate, and has high safety, making it suitable for the room-temperature collection, transportation, and preservation of various biological samples such as blood, saliva, oral swabs, sputum, and tissue homogenates.

The application discloses a tissue homogenizer for detecting bulb vegetables, and relates to the technical field of food detection. The top end of the machine base is fixedly connected with a limiting ring, the inner side of the limiting ring is provided with a lower cylinder, the top end of the lower cylinder is spirally connected with an upper cylinder, the top end of the upper cylinder is spirally connected with an end cover, the inner side of the end cover is rotatably connected with a connecting shaft, the outer side of the connecting shaft is provided with a source suppression mechanism and a defoaming mechanism, the source suppression mechanism is arranged, the surface layer slurry (the region close to the liquid surface) is forced to be pressed to the bottom of the cavity, the design avoids the problems that the traditional cutter swings outward to throw the slurry and impacts the liquid surface, and cuts off the path that the air is rolled into the slurry from the source; the slurry smoothly flows downward along the inclined surface of the cutter, and there is no rolling air phenomenon, so that the air bubbles generated due to the turbulent flow are eliminated from the source.

The invention provides a method for detecting nano-plastics in an organism. The method comprises the following steps: exposing a to-be-detected organism by taking pre-prepared nano-plastics loaded with metal elements as an exposure material; performing homogenization treatment on each tissue of the exposed organism to be detected to obtain tissue homogenate of each tissue; respectively adding an alkaline digestion agent into the tissue homogenate of each tissue; and respectively measuring the digested tissue homogenate of each tissue in a single-particle mode of an inductively coupled plasma mass spectrometer to obtain quantitative and traced detection results of the in-vivo nano plastic of the living body to be detected. According to the method, the to-be-detected organism is exposed by using the metal-loaded nano-plastic, then the tissues of the organism are analyzed one by one, the detection quantitative results of the nano-plastic in the tissues are obtained, the migration process of the nano-plastic in the organism can be deduced according to the physiological relationship between the tissues, and the tracing detection result is obtained.

The invention is applicable to the field of food processing, and provides a preparation method of pig spleen oligopeptide capable of improving immunity, which comprises the following steps: step 1, selecting qualified pig spleen after inspection and quarantine, cleaning, adding a proper amount of clear water for homogenization treatment, then heating to 90-95 DEG C, and keeping the temperature for 15 minutes; step 2, cooling the tissue homogenate after enzyme deactivation to a proper temperature, and carrying out enzymolysis treatment; step 3, adjusting the pH value and temperature of the enzymolysis system to optimal reaction conditions, and continuing enzymolysis; 4, after enzymolysis is completed, standing and precipitating, and separating supernatant enzymatic hydrolysate from residual precipitates; and step 5, performing concentration, dialysis and drying treatment on the enzymatic hydrolysate obtained by separation to finally obtain the pig spleen oligopeptide. The method is simple and convenient to operate, low in cost and high in raw material utilization rate, and the obtained product has the multiple advantages of high purity, easiness in absorption, rich nutrition, strong immunoregulation function and the like.

The invention relates to a construction method of a damp-heat downward flow type erectile dysfunction animal model, which comprises the following steps: S1, from the day of modeling, feeding mice with high-sugar and high-fat feed, and simultaneously providing mixed drinking water containing alcohol and honey, free diet and water; s2, from the eighth day of modeling, feeding the mice in a humid and hot environment; s3, on the 15th day, the 20th day and the 25th day of modeling, a prostate tissue emulsion is adopted to conduct subcutaneous injection on the mouse, a cyclophosphamide solution is synchronously injected into the intraperitoneal cavity, and the prostate tissue emulsion is obtained by emulsifying prostate tissue homogenate and a complete Freund's adjuvant according to the volume ratio of 1: 1; and S4, carrying out comprehensive evaluation on the damp-heat downward flow type erectile dysfunction animal model. Internal and external damp-heat factors are combined with drug induction, the pathogenesis of'damp-heat accumulation in lower jiao 'in traditional Chinese medicine is cooperatively simulated through multiple mechanisms, the manifestation symptom of the animal model is almost consistent with the disease symptom of a clinical damp-heat downward flow type impotence patient, and good scientificity and reliability are achieved.

The invention provides an extraction method of perna viridis foot vesicles, which comprises the following steps: S1, respectively sampling tissues of a near-end region and a far-end region of a foot, and grinding to obtain a foot tissue homogenate; s2, removing tissue fragments and other impurities by adopting a multi-stage filtration mode to obtain a foot vesicle crude extract; and S3, further removing organelles and other impurities by adopting a multi-time centrifugation mode to obtain purified near-end region vesicles and far-end region vesicles of the feet. According to the method, crude extraction of the vesicles is carried out in manners of multiple times of filtration, low-speed centrifugation, filter membrane filtration and the like, and the vesicles can be obtained under the conditions of less centrifugation times, rotation speed and time, so that the extraction process of the vesicles is simplified, and the activity of biomacromolecules in the vesicles is maintained to the maximum extent; the method can be used in the scientific research fields of byssus formation mechanism research, byssus biomimetic material research and development and the like.

The wound healing plaster and the method for its preparation are applicable in medicine. The plaster includes a polymer mixture of polyvinyl alcohol and chitosan, glycerol, and methanol extract from the plant Marrubium peregrinum. The three components are present in the following weight parts: 30-70 parts of polymer mixture, 20-60 parts of methanol extract from Marrubium peregrinum, and 3-25 parts of glycerol. The plasters containing methanol extract from Marrubium peregrinum with a concentration of 4 to 8 mg / cm2 demonstrate very well-expressed wound healing activity, as proven by histological examinations of wound surface samples from rats after euthanasia. The analysis of pro-inflammatory (TNF-α and IL-6) and anti-inflammatory (IL- 10) cytokines in the tissue homogenates, as well as the level of hydroxyproline, showed increased cell proliferation and collagen synthesis compared to control samples that do not contain Marrubium peregrinum extract.

The invention belongs to the field of immunological detection, and discloses an anti-human beta-NGF monoclonal antibody and application thereof. The amino acid sequences of a light chain CDR1, a heavy chain CDR2 and a heavy chain CDR3 of the monoclonal antibody are respectively KSVSTSGYSY, LAS, QHSRELPLT, GFTFSSHW, VRLKSNNYAT and TLNGGAMDH. The amino acid sequences of the light chain CDR1, the heavy chain CDR2 and the heavy chain CDR3 of the monoclonal antibody are respectively KSVSTSGYSY, LAS, QHSRELPLT and GFTFSSHW. The enzyme-linked immunosorbent assay kit constructed by the invention is based on a double-antibody sandwich method, and is combined with a biotin-streptavidin signal amplification system and an enzyme developing amplification system, so that the detection sensitivity and specificity are greatly improved, and low-content samples can be accurately and quickly detected. The kit shows excellent stability and specificity and can be used for quantitatively detecting the content of beta-NGF in samples such as human serum, cell supernatant or tissue homogenate, so that an objective detection basis is provided for diagnosis, curative effect evaluation and pathogenesis research of related diseases.

The utility model discloses a tissue and cell co-culture device, which comprises a culture container, a culture container, a cell culture device and a cell culture device, wherein the lower end of the culture container is step-shaped; a sealing cover is mounted on the upper side of the culture container; a stirring mechanism is mounted on the inner side of the sealing cover; a partition culture frame is mounted on the inner side of the culture container, the partition culture frame comprises a mounting ring, connecting rods are mounted at the lower end of the mounting ring at equal angles, and partition rings are mounted at the lower ends of the connecting rods. The utility model belongs to the technical field of experimental instruments, and achieves the technical effects that the homogenate is attached to the surfaces of cells after being precipitated, so that the absorption of the cells to nutrient substances is hindered, and the growth speed and state of the cells are influenced; the homogenate is prevented from precipitating in a culture container to form a thick layer; and the problems that the contact between the cells and the tissue homogenate is not uniform enough, the interaction complexity is difficult to simulate completely, and the experimental results are inconsistent are solved.

The invention discloses a preparation method and application of sea cucumber polysaccharide, and belongs to the field of food and medicine. According to the method, the sea cucumber intestines are used as raw materials, only the sea cucumber intestines are simply washed in the pretreatment stage, on one hand, polysaccharide loss caused by excessive treatment is avoided, on the other hand, a natural grinding medium is provided for subsequent use of bone paste grinding homogenate, and tissue breaking is facilitated; and then the sand grains and the tissue homogenate are separated through standing. On the basis, polysaccharide extraction is performed by fully utilizing the autolysis characteristic of sea cucumber intestines, and a NaCl solution with a specific concentration is key added in the autolysis stage to inhibit polysaccharide degradation, so that the yield of total sugar and sulfated polysaccharide is increased. Furthermore, a synergistic treatment process of alkaline hydrolysis and enzymolysis is adopted, so that polysaccharide components in the sea cucumber intestines are fully released. Experiments show that the obtained polysaccharide can effectively relieve immunosuppression and intestinal barrier injury caused by cyclophosphamide, and the whole immune function of the body is recovered by adjusting intestinal flora and metabolites thereof.

The utility model relates to the field of instruments for biological tissue homogenate experiments, in particular to a sampling and homogenate integrated instrument and a sampling and homogenate integrated instrument assembly, the sampling and homogenate integrated instrument comprises a tube body, the tube body comprises a main body part and a needle body part, the main body part is provided with a homogenate cavity, the homogenate cavity is provided with an opening, and the needle body part is provided with a needle hole; the homogenizing chamber is used for adding a homogenizing medium into the homogenizing chamber; the needle body part is provided with a tube cavity, the needle body part is used for penetrating into a sampled tissue to enable the sampled tissue to enter the tube cavity, and the tube cavity is communicated with the homogenate cavity to enable the sampled tissue entering the tube cavity to enter the homogenate cavity; the first sealing element is used for sealing the opening; when the sampling and homogenizing integrated appliance assembly is used for sampling and homogenizing, tools do not need to be replaced, operation is simplified, and sample pollution is avoided as much as possible.

The invention discloses a method for analyzing and identifying gastric mucosa tissue strains on the basis of 16S rRNA (ribosomal ribonucleic acid) gene sequences. The identification method comprises the following steps: splitting tissue homogenate and extracting genome DNA (deoxyribonucleic acid); carrying out PCR (Polymerase Chain Reaction) amplification by adopting a 16S rRNA (Ribosomal Ribonucleic Acid) primer, and comparing an amplification product with a gene database to obtain strain classification information of a sample to be detected. The method can be used for rapidly detecting the types of gastric mucosa tissue strains, and the manpower and material resource cost in the microbial species identification process is greatly reduced.

The utility model relates to the technical field of animal experiment equipment, and discloses an animal tissue homogenate operating table which comprises a working table, a transparent protective cover capable of being opened and closed is arranged on the upper portion of the working table, at least one triturating instrument and a porous test tube support are arranged on the working table, the triturating instrument comprises a cup body, a reamer, a cup cover and a main machine, and a temperature control box is arranged on the working table. A placement groove is formed in the temperature control box and can accommodate a cup body of the mashing instrument; an ultraviolet lamp is installed on the top of the inner side of the protective cover, a control panel is arranged on the front face of the workbench, the temperature control box and the ultraviolet lamp are both electrically connected with the control panel, and the temperature control box and the ultraviolet lamp can be controlled to be turned on and turned off through the control panel. The temperature control box is arranged on the working table, a low-temperature environment can be provided for the mashing instrument, a sample is prevented from being degraded or denatured due to overheating of equipment in the homogenizing process, the accuracy and reliability of an experimental result are ensured, a sterile environment can be provided for the working table through the protective cover and the ultraviolet lamp, the operation safety is enhanced, and the risk of cross contamination is reduced.

The application belongs to the field of immunological detection, and provides a double-antibody sandwich ELISA kit for detecting the content of human beta-NGF. The enzyme-linked immunosorbent kit constructed by the application is based on a double-antibody sandwich method, and simultaneously combines a biotin-streptavidin signal amplification system and an enzyme color development amplification system, so that the detection sensitivity and specificity are greatly improved, and low-content samples can be accurately and rapidly detected. The kit exhibits excellent stability and specificity, and can be used for quantitatively detecting the content of beta-NGF in human serum, cell supernatant or tissue homogenate and the like, so as to provide an objective detection basis for the diagnosis, efficacy evaluation and pathogenesis research of related diseases.

The present invention discloses a method for the isolation, purification, staining and counting of Enterocytozoon hepatopenaei. The method comprises the following steps: sample preparation: dissecting the hepatopancreas and intestinal tissues of prawns infected with Enterocytozoon hepatopenaei, preparing a tissue homogenate and diluting it with sterile physiological saline to obtain a diluted homogenate; crude extraction: using differential centrifugation to prepare a crude extract of Enterocytozoon hepatopenaei from the obtained diluted homogenate; the washing medium used in the crude extraction step includes glycerol; purification: using discontinuous sucrose density gradient centrifugation to separate and purify the crude extract of Enterocytozoon hepatopenaei obtained in step S2 to obtain a purified suspension of Enterocytozoon hepatopenaei; staining: staining the Enterocytozoon hepatopenaei in the obtained purified suspension of Enterocytozoon hepatopenaei with a fluorescent brightener; counting: filtering the stained Enterocytozoon hepatopenaei onto a black nuclear pore filter membrane and observing and counting under a high-power fluorescence microscope. The method has the advantages of simple operation, low cost, high pathogen purity, strong specificity of the staining agent and accurate counting.

The invention discloses an extraction and culture method of central nervous system vascular wall cells, and relates to the technical field of cell culture. Microvascular clumps are separated through one-step enzymolysis in combination with two-time centrifugation and four-time natural settling, a large number of high-purity and high-activity vascular wall cells are obtained through in-vitro culture and five-time passage, and the method has the advantages of being easy and convenient to operate, low in cost, high in efficiency, good in cell activity and high in purity. The method comprises the following steps: S1, carrying out enzymatic dissociation on obtained brain and spinal cord tissues by using papain; s2, blowing and beating brain and spinal cord tissues to obtain brain and spinal cord tissue homogenate; s3, adding bovine serum albumin into the brain and spinal cord tissue homogenate, and centrifuging twice to obtain a low-purity microvascular plexus; s4, naturally settling the low-purity capillary plexus for four times to obtain a high-purity capillary plexus; s5, inoculating the high-purity capillary plexus into a culture plate; s6, performing in-vitro culture to enable the cells to climb out of the capillary plexus to obtain vascular wall cells; and S7, carrying out passage for five times to obtain the high-purity vascular wall cells.

The invention relates to a method for preparing ROS (reactive oxygen species) response carbon dots by a one-step microwave method, which comprises the following steps: adding ultrapure water into citric acid, betaine and 4-hydroxyphenylboronic acid, and uniformly mixing to obtain a uniform precursor solution; reacting for 3 to 5 minutes under the condition that the power of a microwave oven is 700 to 800 W; after the reaction, naturally cooling, filtering, transferring into a dialysis bag, and dialyzing at room temperature by taking deionized water as a dialysate; and freezing and drying. The core reaction of the one-step microwave method only needs 3-5 minutes, the total preparation period is less than or equal to 30 hours, and high-temperature and high-pressure equipment is not needed; the raw materials are laboratory conventional reagents, the cost is low, and batch production can be realized; the kit has specific response to H2O2, the detection limit is less than or equal to 0.8 mu M, and the anti-interference capability is strong; the carbon dots can be used for quantitative detection of H2O2 in a biological system (such as cells and tissue homogenate), can also be used for ROS content analysis in an environmental water sample and a food sample, are good in biocompatibility and strong in anti-interference capability, and can be suitable for ROS detection inside and outside an organism and in the environmental system.

The invention belongs to the field of nucleic acid detection, and discloses a method for detecting oligonucleotide drugs based on an MSD (Meso Scale Discovery) platform and application of the method. According to the method, high-sensitivity quantitative detection is realized by designing a specific probe system: a capture probe and a detection probe are synergistically hybridized with target oligonucleotide to form a compound, the compound is fixed on a streptavidin-coated MSD plate, an anti-digoxin-SULFO-TAG antibody is combined, and a signal is detected by an electrochemical luminescence method. The innovation points are as follows: LNA modification enhances the stability and specificity of the probe, and the use amount of the probe is reduced by 50%; the process of a non-enzyme reaction system is simplified to 4 hours; sample treatment is optimized (RNALater is added to inhibit RNA enzyme degradation), and the method is suitable for complex matrixes such as serum and tissue homogenate. The sensitivity reaches 0.4 pM, the linear range spans five orders of magnitude, high specificity and high flux are achieved, and efficient and reliable technical support is provided for research and development of oligonucleotide drugs, pharmacokinetic research and clinical tests.