528 results about "Immunofluorescence" patented technology

There is proposed a method for detecting a presence of a fluorescence pattern type on an organ segment via immunofluorescence microscopy and digital image processing. The steps comprise: provision of the organ segment, incubation of the organ segment with a liquid patient sample, incubation of the organ segment with secondary antibodies which have been labelled with a fluorescent dye, acquisition of a fluorescence image of the organ segment in a colour channel corresponding to the fluorescent dye, and provision of the fluorescence image to a neural network. What takes place by means of the neural network is simultaneous determination of segmentation information through segmentation of the fluorescence image and, furthermore, of a measure of confidence indicating an actual presence of the fluorescence pattern type. What further takes place is determination, on the basis of the previously determined segmentation information, of at least one sub-area of the fluorescence image that is relevant to formation of the fluorescence pattern type, determination, on the basis of the previously determined at least one sub-area, of validity information indicating a degree of a validity of the measure of confidence, and output of the measure of confidence depending on the validity information.

The invention provides a multi-modal kidney pathological picture processing method and device, electronic equipment and a medium. The method comprises the following steps: acquiring an immunofluorescence picture included in a to-be-processed multi-modal kidney pathological picture and a PAS staining pathological picture corresponding to the immunofluorescence picture; inputting the immunofluorescence pictures into a pre-trained first network model to perform nephropathy type classification to obtain nephropathy types; judging whether the nephropathy type output by a pre-trained first network model belongs to a preset nephropathy type or not; inputting a PAS staining pathological picture corresponding to the immunofluorescence picture into a pre-trained second network model for pathological structure identification when the immunofluorescence picture belongs to the preset nephropathy type, and outputting a label for the identified pathological structure; and adding an Oxford typing label to the to-be-processed multi-modal kidney pathological picture based on a label and a pathological structure output by a pre-trained second network model. According to the scheme, the nephropathy type can be automatically distinguished, the Oxford typing label is added, the processing speed is high, and the accuracy rate is high.

The invention belongs to the technical field of tumor micro-immune environments, and particularly relates to a kit for detecting a tumor micro-environment, a detection method and application, the kit comprises rare metal element labeled monoclonal antibodies, the rare metal element labeled monoclonal antibodies comprise 115In-CD20, 139La-Histone H3, 142Nd-CD56, 144Nd-CD14, 146Nd-Vimentin, 148Nd-Pan-CK, 151Eu-CD31, 156Gd-CD4, 158Gd-E-Cadrin, 159Tb-CD68, 162Dy-CD8a, 166Er-CD45, 167Er-CD15, 170Er-CD3, 171Yb-CD11c and 209Bi-alpha-SMA, and the rare metal element labeled monoclonal antibodies comprise The invention can be used for simultaneously detecting and researching the quantity, positioning and spatial topological relation of each cell component in a tumor microenvironment, and provides a multi-immunofluorescence detection kit and method for ubiquitous epithelial differentiation-derived tumors based on an imaging mass spectrum flow technique. A plurality of biomarkers can be analyzed at the same time by using multiple immunohistochemical staining and rare earth metal tag signal detection technologies, so that the aim of performing high-dimensional analysis on a tumor microenvironment is fulfilled according to cell composition, functional state and cell-cell space interaction relationship.

The invention provides a storage system for immunofluorescence slide glass and an intelligent temperature and humidity coupling control method thereof. The system comprises a closed sample cabin, a controller, a passive buffer layer and other structures, and the temperature and humidity in the cabin can be stably controlled in a dynamic environment. The temperature and humidity in the cabin and the temperature of the key cold surface are monitored in real time through the sensing unit, the local dew point is calculated, and temperature and humidity control is executed. And the controller executes a layered gradient coupling control algorithm according to the dynamic dew point safety margin to ensure accurate control of temperature and humidity. The passive buffer layer is of a composite structure of a phase change material and a moisture absorption material, and the temperature and humidity are adjusted through solid-liquid phase change and molecular adsorption. The system can effectively improve the storage stability and safety of the immunofluorescence slide, and is suitable for sample storage management in industries such as life science and biological pharmacy.

The invention belongs to the technical field of medical diagnosis, and discloses a quantum multicolor immune digital pathological diagnosis analysis system, which comprises the following steps: acquiring high-resolution digital images of an immunohistochemical or immunofluorescence slice and a slide, and analyzing multicolor signal distribution in the images to obtain a quantum chromatographic image set; pixel-level color separation and intensity quantization are carried out on the quantum chromatographic image set, and a quantum intensity index set of each pixel is generated; performing multi-target color overlay analysis and post-translational modification protein specificity evaluation on the quantum intensity index set to construct a multi-color modification index set; identifying a single cell boundary and a multi-cell community based on the multicolor modification index set, and extracting a cell morphological parameter and a spatial adjacency relation to obtain a cell quantum analysis set; performing consistency correction and traceable recording by applying an automatic quality control mechanism, and generating a digital diagnosis report; the accuracy of accurate qualitative and quantitative analysis of immunohistochemistry and immunofluorescence is greatly improved.

The invention provides a full-automatic indirect immunofluorescence interpretation method and system, a storage medium and a program product, and relates to the technical field of indirect immunofluorescence detection.The method comprises the steps that when identification information of a target sample is obtained, a plurality of sealed antigen slides are placed in slide carrying groove positions of a bearing platform in batches, and the target sample is obtained; driving the bearing platform through the transmission mechanism to sequentially move the plurality of antigen slides to the imaging area, and performing positioning and fixing operation on the antigen slides entering the imaging area by using the positioning and clamping device; sequentially carrying out multi-view scanning on the antigen slides entering the imaging area to obtain multi-view fluorescence images, and splicing the fluorescence images to generate a fluorescence panoramic image; performing feature extraction on the panoramic image, determining a fluorescence karyotype category and a target fluorescence intensity value, and determining an antibody titer according to the category, the intensity value and dilution information; and performing association mapping on the titer, the category and the panoramic image according to the identification information to generate a fluorescence interpretation result.

The invention relates to the field of molecular cytogenetics and potato ploidy breeding, in particular to a potato ploidy identification method for marking and counting chromosome centromere based on a potato CENH3 antibody and a ploidy detection kit. According to the method, the chromosome centromere is marked by specifically recognizing the antibody of the potato centromere nucleosome core histone CENH3 and combining the immunofluorescence technology, so that the accurate counting of the chromosome centromere is realized at the single cell level, and the ploidy of potatoes is judged. The method comprises the following steps: preparing a leaf cell suspension, sequentially incubating a potato CENH3 antibody and a labeled secondary antibody after immobilization, and observing and counting by a microscope after redyeing. The method has the advantages of high species specificity, high throughput and low cost, is not limited by special material taking parts of root tips or shoot tips and cell division periods, can accurately identify the uploid and aneuploid of potatoes, and provides an efficient tool for potato germplasm resource evaluation and cross breeding.

The invention relates to the technical field of life science, in particular to a method for constructing an atrophic gastritis-based SPEM lesion model, which comprises the following steps: S1, constructing an Slc7a11 conditional knockout mouse strain: carrying out cage hybridization on an Slc7a11 flox / flox mouse and an ATP4b-Cre transgenic mouse, and knocking out a gastric wall cell specific Slc7a11 gene; s2, carrying out genotyping identification; the identification method based on the atrophic gastritis SPEM lesion model and the construction method based on the atrophic gastritis SPEM lesion model comprise the following steps: S1, pathological model verification: taking a mouse gastric mucosa tissue slice, quantitatively analyzing the proportion of an SPEM region by adopting an immunofluorescence double-standard technology, and confirming Slc7a11 knockout induced SPEM pathological characteristics in combination with the existence of SPEM in the gastric mucosa; and S2, long-term pathology evolution monitoring. Through gastric wall cell specific Slc7a11 gene knockout and multiple PCR identification technologies, the initial response SPEM state after gastric mucosa damage is successfully simulated, and the kit is used for researching atrophic gastritis pathological mechanisms and evaluating drug intervention.

The present invention relates to RyR1 antigens, antigen expression genes, antigen expression vectors, cell lines and uses. The present invention provides two effective antigens for detecting anti-RyR1 autoantibodies: MGTR1 and MGTR2, which have the characteristics of small molecule size and good expression. Expression plasmids carrying the MGTR1 and MGTR2 genes are transfected into cell lines to prepare cell-based detection materials. The prepared detection materials have good sensitivity, specificity and stability for RyR1 antibodies. The indirect immunofluorescence method using cells is used to screen for anti-RyR1 autoantibodies in MG patients and MG patients with thymoma. Based on the RyRl cell detection matrix material, a two-color fluorescence system is adopted, and its co-localization effect is good. Therefore, it has good sensitivity, specificity and stability. The RyR1 cell detection matrix of the present invention can meet the needs of auxiliary clinical diagnosis.

The invention discloses an anti-soluble transferrin receptor monoclonal antibody as well as a preparation method and application thereof, and belongs to the technical field of antibody detection. According to the antibody, an L101-F760 amino acid sequence of human TfR protein is used as an immunogen, specific screening of B cells, reverse RNA (Ribonucleic Acid) into cDNA (Complementary Deoxyribose Nucleic Acid) through RT-PCR (Reverse Transcription-Polymerase Chain Reaction), then homologous recombination is performed to construct heavy and light chain vectors, and finally multiple rounds of screening are performed to obtain the purified sTfR monoclonal antibody, so that the sTfR protein can be specifically recognized, and the antibody has the advantages of high specificity and high sensitivity and can be widely applied to the field of immunotherapy. The natural sTfR protein in human serum can be recognized. The kit is applied to the detection and screening fields of immunohistochemistry, immunofluorescence chemistry, western blot, co-immunoprecipitation, indirect ELISA, flow cytometry and the like, especially has low background and no interference of non-specific binding impure protein during immunohistochemistry detection, can obtain more accurate detection and evaluation results, avoids interference of false positive and false negative, and has high detection accuracy. The method has great application value in the aspects of basic scientific research and clinical pathology detection.

The invention discloses an IgA nephropathy prognosis method, system and device based on multi-modal data, and belongs to the technical field of image data processing, the method comprises the following steps: collecting historical data including clinical data and an IgA immunofluorescence map; based on a visual identification method, pathological features are extracted from the IgA immunofluorescence image; screening clinical characteristics; based on a machine learning method, the training set is trained according to pathological features and clinical features, a prognosis model is obtained, and the prognosis model is used for IgA nephropathy prognosis. On the basis of a computer vision method, pathological features are extracted from an IgA immunofluorescence map, and the prognosis of the IgA nephropathy is predicted in combination with clinical features, so that automatic prediction is facilitated, mistakes and omissions caused by artificial naked eye recognition are avoided, stable prediction ability is expressed, and multi-modal data fusion reflects a gain effect on long-term prognosis prediction.

The invention relates to the field of biology, and provides a human immortalized pancreatic cancer fibroblast as well as a preparation method and application thereof.The human immortalized pancreatic cancer fibroblast comprises three cell strains which are all primary fibroblast cells extracted from tumor tissues of pancreatic ductal adenocarcinoma patients, the human pancreatic cancer fibroblast cell strain CAFPC-2209131, the human pancreatic cancer fibroblast cell strain CAFPC-2209132 and the human pancreatic cancer fibroblast cell strain CAFPC-2303221 are classified and named as the human pancreatic cancer fibroblast cell strain CAFPC-2209131, the human pancreatic cancer fibroblast cell strain CAFPC-2209132 and the human pancreatic cancer fibroblast cell strain CAFPC-2303221 respectively, the human pancreatic cancer fibroblast cell strain CAFPC-2209132 and the human pancreatic cancer fibroblast cell strain CAFPC-2303221 are all Cell STR identification and immunofluorescence detection prove that the cell strain is a fibroblast cell strain. After the tumor-associated fibroblast and pancreatic cancer cells are co-cultured, the proliferation ability of the tumor cells is obviously enhanced, and the sensitivity of the tumor cells to a chemotherapeutic drug gemcitabine is reduced. A new thought is provided for diagnosis and treatment of pancreatic cancer, and based on the pancreatic cancer in-vitro co-culture model established in vitro, the application has important significance for researching the effect and related mechanisms of fibroblasts in the pancreatic cancer tumor microenvironment.

The invention discloses an application of an AP2-F2 protein polyclonal antibody as a cryptosporidium female gamete positioning tag antibody. The AP2-F2 protein of cryptosporidium parvum is subjected to truncated prokaryotic expression, protein is purified to immunize an ICR mouse, a polyclonal antibody is prepared, ELISA and WB detection of antibody titer are carried out to prove that the polyclonal antibody is effective, then indirect immunofluorescence detection of the antibody is carried out to determine that the polyclonal antibody is located on the female gametes of cryptosporidium parvum, and the polyclonal antibody can be used as a label to locate the female gametes of cryptosporidium parvum. The method plays an important role in localization research of unknown protein of cryptosporidium.

The invention relates to the fields of epigenetic regulation and tumor treatment, and aims to evaluate that the cell distribution of the mutant is consistent with that of H3 by constructing Bas-2B (H3K9R) and Bas-2B (H3K18R) stably transfected cell strains and analyzing the cell positioning condition of the H3K9R / H3K18R mutant through an immunofluorescence technique. According to the present invention, a Western blotting technology is adopted to detect the H3K9 / H3K18 lactic acid modification level of Bas-2B (H3K9R) and Bas-2B (H3K18R) cells compared to control cells so as to verify the influence of the H3K9R / H3K18R mutant on the H3K9 / H3K18 lactic acid modification; after Bas-2B (H3K9R), Bas-2B (H3K18R) cells and control cells of the Bas-2B (H3K9R) and the Bas-2B (H3K18R) cells are subjected to nickel compound exposure treatment, the influence of the H3K9R / H3K18R mutant on the nickel compound induced lung cancer is evaluated through a soft agar cloning formation experiment. An AGS cell line stably transfecting H2BK16R is constructed, and the cell localization condition of the H2BK16R mutant is analyzed through an immunofluorescence technology. According to the present invention, a Westernbloting technology is adopted to detect the lactic acid modification level of AGS (H2BK16R) cells so as to verify the influence of the H2BK16R mutant on the H2BK16 lactic acid modification; the influence of the H2BK16R on the malignant proliferation ability of the gastric cancer is researched through a soft agar clone formation experiment.

The invention relates to the technical field of general image data processing or generation, in particular to a multi-view indirect immunofluorescence medical image classification interpretation method and system, and the method comprises the steps: mapping an ethanol view, a formaldehyde view and an HEp-2 view to a feature space, and converting into a feature sequence; performing position coding on the feature sequence to obtain position information; based on the feature sequence with the position information, modal identification information is added, and an identification feature sequence is determined; based on the identification feature sequence, executing multi-modal information fusion, and determining a multi-modal feature sequence; based on the multi-modal feature sequence, combining an attention mechanism to determine a long-distance dependency relationship and correlation between the data; and based on the attention mechanism model, outputting the probability of each category to execute classification interpretation. According to the method, the type of the anti-neutrophil cytoplasm antibody fluorescence model can be adaptively and comprehensively judged in combination with the characteristics of fluorescence brightness, forms and the like of different views, and interference existing in a single view is avoided.

The application discloses a kind of multiple immunofluorescence detection method, equipment and medium based on space reconstruction, it is related to histopathological analysis technical field, including, preparation tissue sample and join fluorescent reference microbead to obtain continuous section;To the tissue sample cycle pretreatment, and utilize fluorescent reference microbead to carry out intensity normalization and bleaching time shift correction, obtain correction image sequence;Spectral unmixing is carried out under three-dimensional voxel coordinate system and introduces space regular constraint, obtain voxel level fluorescence intensity vector;Combining nuclear dye signal and fluorescent reference microbead, using rigid and non-rigid registration and supplemented with three-dimensional deconvolution, reconstruct high-resolution voxel stack and segment cell voxel set;The adjacent relationship and colocalization index of different phenotype cells are calculated, and detection result is generated.The application realizes high-resolution tissue and cell reconstruction under three-dimensional space, improves spatial continuity and accuracy, and improves the stability and accuracy of voxel level fluorescence signal quantification.

The invention belongs to the technical field of biomedicine, and discloses an antinuclear antibody IgG indirect immunofluorescence detection method and system, and the detection standardization degree and the result reproducibility are significantly improved through the full-automatic system design in combination with precise mechanical transmission and an automatic sample adding device. Through an optimized FITC labeling system and a refined washing procedure, non-specific binding is remarkably reduced, and meanwhile, the binding efficiency of the antinuclear antibody and the secondary antibody is improved, so that the detection sensitivity is improved. Through the design of the automatic sample adding and sheet sealing device, the sample treatment efficiency is greatly improved, and simultaneous detection of large-scale samples can be realized. Through combination of a high-resolution imaging system and an intelligent image analysis algorithm, morphological characteristics and distribution rules of fluorescence signals are automatically extracted, and the accuracy and objectivity of a detection result are greatly improved.

The invention relates to the technical field of biology, and particularly discloses a construction method and application of an endometrial epithelium organ fibrosis model. According to the method, human endometrial epithelial cells are extracted, and an endometrial epithelial organ is established by adopting a Matrigel three-dimensional culture system. Afterwards, the organoid is induced to form endometrial phenotypes in different physiological cycles through hormone stimulation, and a ferroptosis inducer FIN56 is added in the culture process to induce fibrosis. The expression of GPX4, Fibronectin and p-Smad2 / 3 is detected through Western blot and immunofluorescence, and the successful construction of the fibrosis model is verified. The invention provides an in-vitro model capable of simulating endometrial fibrosis pathological characteristics of a patient with PCOS (polycystic ovarian syndrome), the in-vitro model can be used for mechanism research and drug screening of PCOS-related endometrial lesions, and a new research tool and an intervention strategy are provided for treatment of PCOS endometrial fibrosis.

Methods and systems for analysing the cellular composition of a sample are described, comprising: providing an image of the sample in which a plurality of cellular populations are associated with respective signals and classifying a plurality of query cells in the image between a plurality of classes corresponding to respective cellular populations in the plurality of cellular populations. This is performed by providing a query single cell image to an encoder module of a machine learning model to produce a feature vector for the query image, and assigning the query cell to one of the plurality of classes based on the feature vector for the query image and feature vectors produced by the encoder module for each of a plurality of reference single cell images. The machine leaning model comprises: the encoder module, configured to take as input a single cell image and to produce as output a feature vector the single cell image, and a similarity module configured to take as input a pair of feature vectors for a pair of single cell images and to produce as output a score indicative of the similarity between the single cell images. Thus, the machine learning model can be obtained without the need for an extensively annotated dataset. The methods find use in the analysis of multiplex immunohistochemistry / immunofluorescence in a variety of clinical contexts.

The invention discloses a serum migration body identification method which comprises the following steps: extracting a serum migration body from serum by performing multiple centrifugal treatment on the serum; carrying out immunofluorescence staining treatment on the serum migration body to obtain an immunofluorescence stained serum migration body; and carrying out imaging treatment on the serum migration body subjected to immunofluorescence staining by utilizing a super-resolution fluorescence microscope SIM to identify the serum migration body. Whether an extracellular vesicle in serum is a migration body or not is identified from the two aspects of biochemistry and shape, so that the defect that the serum migration body is identified only from the shape in the prior art is overcome.

The invention relates to the technical field of optics, and discloses a visualization method of a biological tissue three-dimensional full-information atlas, which comprises the following steps of: acquiring image data, electron microscope data, mechanical data, protein distribution, omics information and histological staining images of biological tissues by using various biological visualization technologies, and correspondingly processing the acquired data to obtain a three-dimensional full-information atlas of the biological tissues; finally, organization information fusion and digital display are achieved. According to the method, a cross-scale feature correlation algorithm framework is innovatively established, and multi-source heterogeneous data fusion is performed on medical image topological data (CT / MRI), ultrastructure scanning data (SEM / TEM), molecular positioning information (immunofluorescence, immunohistochemistry and enzyme immunoassay), omics maps (space transcriptome / proteome) and high-resolution tissue staining data. The data island dilemma caused by traditional single-mode analysis is broken through, and full-scale visualization from nanoscale molecular distribution to centimeter-level organ tissue structure-function relationship is realized for the first time.

The invention discloses a fluorescence coding probe and a single cell immunoblotting method. The nucleic acid tag comprises oligonucleotide chains f1, f2... fn, n is greater than 2, each oligonucleotide chain contains a fluorophore and a quenching group, partial sequences of the next oligonucleotide chain and the previous oligonucleotide chain are complementarily paired, and the quenching group of the next oligonucleotide chain quenches the fluorophore of the previous oligonucleotide chain. Also disclosed is a quenching tag comprising a complementary strand capable of complementary pairing with an oligonucleotide strand in a nucleic acid tag. The invention discloses a fluorescent coding probe comprising a nucleic acid label, a preparation method of the fluorescent coding probe and application of the fluorescent coding probe in a multi-target single-cell immunoblotting technology, an immunofluorescence technology and an immunohistochemical staining technology. According to the fluorescence-coded multi-target single-cell immunoblotting method disclosed by the invention, multi-target protein detection can be carried out by utilizing limited fluorescence channels, the signal loss is small, and the expansibility is high.

Example systems and methods for generating virtual immunofluorescence stains for tissue samples are provided. A computing device receives a slide image of a target tissue sample of a particular tissue type. The computing device selects a first trained machine learning (ML) model to generate virtual immunofluorescence (IF) stains of a first type for the particular tissue type based on a user input. The first trained ML model is trained at least based on a first set of stain images of a plurality of training tissue samples with stains of the first type. The computing device generate a virtually stained image of the target tissue sample with the virtual IF stains of the first type using the first trained ML model. The computing device displays the virtually stained image of the target tissue sample with the virtual IF stains of the first type.

The invention relates to a circulating tumor cell enrichment, separation and dyeing integrated system and application thereof.The circulating tumor cell enrichment, separation and dyeing integrated system is composed of a bottom plate module, a platform module, a multi-pipette module, a power module and a shell module; the platform module comprises a sample tube rotating and uniformly mixing module, a magnetic adsorption module, a reagent module, a waste liquid pool module and a gun head module, and the sample tube rotating and uniformly mixing module, the magnetic adsorption module and the reagent module are fixedly mounted on a bottom plate I in sequence. The CTC separation and dyeing integrated system is developed on the basis of the immunomagnetic bead technology, enrichment separation and immunofluorescent dyeing of CTC can be automatically completed by running a program, manual operation errors are effectively avoided through a programmed process, and the CTC separation and dyeing integrated system is particularly suitable for clinical CTC detection. The system is also suitable for other enrichment and separation applications based on an immunomagnetic bead technology, including but not limited to enrichment and separation of biological macromolecules such as cells, exosomes, proteins and the like.

The utility model belongs to an immune technology, and particularly relates to a single-molecule immunofluorescence analyzer which comprises a light source and a convergent lens, an optical channel is formed in the guide piece; the moving part is arranged on the outer side of the guide part and moves along the guide part under driving; the fixed part is used for fixing the convergent lens on the movable part, and light penetrating through the convergent lens penetrates through the optical channel; the transmission mechanism is used for converting rotation of the rotating mechanism into rotation of the moving part. The utility model has the advantages of high adjustment precision and the like, and is used in optical focusing.

The invention provides an immunofluorescence detection method and system for dynamically detecting mouse uterus structure development based on multiple time points, and immunofluorescence staining comprises the steps of sheet unfolding, dewaxing, repairing, cell membrane punching, sealing, primary antibody treatment and secondary antibody treatment. The system comprises a sample pretreatment control module, a multi-channel confocal microscope imaging module and a data analysis module integrating fluorescence intensity calculation, Pearson co-localization analysis and a Western blot data integration algorithm. Through a multi-time-point and multi-index immunofluorescence detection method, dynamic and systematic data support is provided for mouse uterus development mechanism research, and an application basis is provided for fundamental research of reproductive biology and construction of related disease models.

The invention discloses an immunofluorescence imaging method for driving tyramine signal amplification based on chemiluminescence and application thereof, and relates to the technical field of biomedical imaging. The immunofluorescence imaging method comprises the following steps: carrying out first incubation on a to-be-detected sample and a peroxidase-labeled antibody; performing second incubation on the to-be-detected sample after the first incubation and a tyramine substrate containing a fluorophore under a dark condition; adding a chemiluminescent substrate solution into the to-be-detected sample after the second incubation, and carrying out image acquisition; wherein the antibody can be specifically bound with a target protein; the tyramine substrate can generate active free radicals under the catalysis of peroxidase, so that the active free radicals are covalently cross-linked with tyrosine residues; the chemiluminescent substrate solution comprises hydrogen peroxide and luminol. The immunofluorescence imaging method improves imaging signal and fluorescence stability, effectively reduces background interference and phototoxicity, is suitable for long-time stable fluorescence immunoimaging, and is especially suitable for high-sensitivity detection of low-expression protein markers.

The invention provides a toxoplasma gondii gene deletion strain with an immune protection effect and application of the toxoplasma gondii gene deletion strain, and belongs to the technical field of veterinary parasitology, veterinary immunology and parasite molecular biology. According to the toxoplasma gondii gene deletion strain, a gamma-glutamyl hydrolase gene is deleted. The invention also provides a method for constructing the insect strain. The method comprises the following steps: constructing a circular plasmid pSAG1-Cas9-U6-sgGGH; constructing a repair template containing upstream and downstream homologous arms of the gamma-glutamyl hydrolase gene and a DHFR resistance gene; co-transfecting the annular plasmid and the repair template to a toxoplasma gondii ME49 strain; performing drug screening on pyrimethamine to obtain positive monoclone; and confirming that the GGH gene is completely knocked out through PCR (Polymerase Chain Reaction) detection, sequencing analysis and immunofluorescence detection. The toxoplasma gondii gene deletion strain can provide a remarkable protection effect on toxoplasma gondii infection and is high in safety.

The invention discloses application of eugenin in preparation of a medicine for treating cerebral arterial thrombosis, and belongs to the technical field of medicines. An in-vivo middle cerebral artery occlusion model is adopted, after reperfusion is conducted for 24 h, the cerebral infarction volume and movement coordination function defect of a mouse are detected to evaluate the treatment effect of eugenin, and activation of microglial cells and survival of neuronal cells are observed through immunofluorescence to clarify the action way of eugenin. Pharmacological activity shows that eugenin can obviously reduce the cerebral infarction volume of a model mouse, improve pathological damage of cells and promote recovery of a motion coordination function, and the action way of eugenin is to promote survival of neuronal cells and inhibit activation of microglial cells. Research results indicate that eugenin has the effect of resisting cerebral arterial thrombosis and can be used as a candidate medicine for developing the cerebral arterial thrombosis resisting medicine.

The invention provides an organoid membrane protein and intracellular protein immunofluorescence in-situ co-staining method, which comprises the following steps of: sealing and fixing organoid glue drops, adding a mixed solution of membrane protein primary antibody and a Dune phosphate buffer solution containing bovine serum albumin for incubation, performing membrane permeation by using a permeation agent, sealing and washing, and drying to obtain the organoid membrane protein and intracellular protein immunofluorescence in-situ co-staining product. Adding a mixed solution of an intracellular protein primary antibody and a Dune phosphate buffer solution containing bovine serum albumin into the cell plate where the transparent organ-like glue drops are located, incubating and washing, then adding a mixed solution of a membrane protein and a fluorescent secondary antibody of the corresponding species of the intracellular protein, incubating and washing in a dark place, and finally obtaining the membrane protein / intracellular protein fluorescent antibody. And finally, adding a mixed solution of DAPI and a Duncheng phosphate buffer solution, and incubating and washing in a dark place again to obtain the membrane protein and intracellular protein immunofluorescence in-situ co-dyed organoid. According to the method, high-quality and high-fidelity membrane protein and intracellular protein in-situ co-dyeing is successfully realized, the integrity of a three-dimensional structure and the high efficiency of an experimental process are taken into consideration, and the method is convenient to operate, high in repeatability and good in universality.