9 results about "DAPI" patented technology

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.

The invention relates to the field of CAR-T cell concentration detection, in particular to a method for detecting CAR-T cells through combination of microfluidics and immunofluorescence. Through immunomagnetic bead enrichment and micro-fluidic chip partitioning, CAR-T cells are separated from a blood sample through immunomagnetic beads, then labeling is carried out or enhanced by adding a labeling antigen, anti-TCR-PE and DAPI, three-channel fluorescence labeling is achieved, quantitative analysis can be carried out on the number, the cell morphology and the function of the CAR-T cells through automatic immunofluorescence imaging, and the accuracy of CAR-T cell detection is improved. And the detection limit reaches 0.001%.

The invention discloses a method for predicting the curative effect of triple negative breast cancer immunotherapy by spatially quantizing CD39 + CD103 + CD8 + T cells, and relates to the technical field of biology.The method comprises the following specific steps that S1, an FFPE tumor tissue sample of a TNBC subject before immunotherapy is obtained, and a continuous section with the thickness of 3-5 microns is prepared and attached to a positive charge glass slide; s2, staining the section by using a combination containing CD8 / CD103 / CD39 / Pan-CK antibodies through a TSA sequential staining method, and then re-staining cell nucleuses by using DAPI; according to the method, spatial quantitative analysis is performed on a specific CD39 + CD103 + CD8 + T cell subset, a percentage index of target cells in CD8 + T cells and an average distance index of the target cells and nearest Pan-CK positive cells in a cancer nest region are set, and the target cells are determined according to comparison between the two indexes and a preset threshold value. According to the method, whether a subject is a potential benefit crowd treated by an anti-PD-1 / PD-L1 immune checkpoint inhibitor or not is accurately judged, a standardized technical means is provided for clinically screening the potential benefit crowd for immunotherapy, the accuracy and effectiveness of triple-negative breast cancer immunotherapy are effectively improved, and invalid treatment is reduced.

A spatial in situ sequencing method, belonging to the field of biology, is proposed. It utilizes an electric field-assisted directed migration of mRNA and in-situ capture with primers on a microarray surface to enrich tissue and release mRNA. In-situ reverse transcription generates covalently fixed cDNA, ensuring high positional stability during multiple rounds of hybridization and imaging. After reverse transcription, tissue is digested to remove tissue, reducing spatial hindrance and background interference, while the cDNA remains at its original coordinates due to covalent anchoring. Combining coding probe hybridization and RCA, single-molecule-level signal amplification and recognition are achieved. Through decoding and single-cell segmentation, transcripts are mapped to their respective cells, constructing a single-cell resolution spatial gene expression map. This method does not rely on multi-round DAPI mapping or other endogenous morphological marker-based multi-cycle image registration methods, making it suitable for high-throughput spatial in situ sequencing and significantly improving robustness and versatility in complex imaging scenarios such as thick tissue sections and low signal-to-noise ratios. It is applicable to high spatial resolution, high-throughput spatial transcriptome research.

This invention discloses a method for detecting Candida albicans based on compound fluorescent staining and deep learning, belonging to the field of microbial detection technology. The method first synthesizes a novel NHC-triazolium salt fluorescent molecule, which is then compounded with calcium fluorescent white and DAPI to form a specific staining solution. After treating the sample with this staining solution, multi-channel fluorescence imaging is performed and images are acquired. Simultaneously, ITS sequencing is performed on the samples. Based on the sequencing results, the images are classified and bacterial cells are manually cropped to construct a labeled dataset. A convolutional neural network model is trained using this dataset to learn to recognize the unique "multi-fluorescent fingerprint" presented by Candida albicans stained with the compound staining solution. In application, the same staining and imaging are performed on the test samples, and the trained model automatically analyzes the images to achieve detection. This invention significantly improves the specificity of detection through the synergy of chemical staining and artificial intelligence. Experiments show that compared with traditional single staining or unstained methods, this method improves the specificity of Candida albicans recognition by 37% and 52%, respectively, and has the advantages of being fast, accurate, and highly automated.

The application belongs to the field of cell culture. The application provides a culture medium of rat amniotic membrane mesenchymal stem cells and a culture method of rat amniotic membrane mesenchymal stem cells. The culture medium is a complete culture medium, and the culture method is as follows: the rat amnion is cut into tissue blocks smaller than 2mm 2 , is uniformly laid in a culture dish with a spacing of 0.3-0.8cm, complete culture medium is slowly added after the tissue blocks are adhered, the culture medium is supplemented after 12h-36h, the primary medium replacement is performed after 48h-72h of culture, the impurity cells are removed, and the cells are subcultured when the cell fusion degree reaches 80%-90%. The light microscope observation and identification result show that the rat amniotic membrane mesenchymal stem cells cultured by the application basically conform to the cell morphology of rat amniotic membrane mesenchymal stem cells, the CD90&DAPI immunofluorescence identification result shows that the cell purity is greater than or equal to 90%, the cells have the characteristics of stem cells, and the culture method is efficient in amplification.

The invention discloses a method for observing the shape and number of synapses in a dentate gyrus region based on image analysis, and relates to the technical field of neurobiology. The method comprises the following steps: S1, tissue treatment: after an experimental animal is anesthetized, pouring a buffer solution and a stationary solution into the heart in stages, separating hippocampus tissue, trimming and fixing, pre-cooling, slicing and preserving at low temperature; s2, dyeing and marking: performing targeted marking on the dentate gyrus area of the sea horse by adopting DiI particles through a fine tungsten steel through needle, and preserving at low temperature after incubation; s3, fixing and mounting: fixing for the second time, and mounting with an anti-fluorescence quenching agent after DAPI nuclear dyeing; s4, confocal imaging is carried out, exclusive parameter layer scanning is set, and qualified dendritic spine images are screened; and image quantitative analysis can be further carried out, and accurate quantification is realized according to classification standards. The method is standard in operation, greatly improves sample quality and experimental result credibility, optimizes experimental efficiency and controllability and reduces technical threshold by means of targeted marking and standardized processes, and is suitable for relevant basis and application research of the hippocampal synapses.

The invention relates to the technical field of biomedicine, in particular to an automatic analysis and risk assessment system and method for peripheral blood endothelial progenitor cells, and the method comprises the following steps: collecting a peripheral blood sample and clinical data of a subject; the method comprises the following steps: enriching CD34 + cells from a peripheral blood sample based on microfluidic-immunomagnetic bead enrichment, carrying out VEGFR2 and DAPI immunofluorescent staining on the enriched cells, and obtaining a multi-channel fluorescent digital image; analyzing the multi-channel fluorescent digital image, identifying and counting VEGFR < 2 + > and DAPI < + > double-positive cells to obtain an EPC counting result; inputting the EPC counting result and clinical data of the subject into a multi-modal AI risk assessment model to obtain a risk assessment result; the health management suggestion information is formulated based on the risk assessment result, so that the model can have better suitability with the target subject, and the accuracy of risk assessment is improved.

The present application relates to pathological image processing technical field, provide a kind of tissue section image and DAPI staining image registration method, comprising: tissue section image and DAPI staining image are carried out gray processing, and according to same work magnification is enlarged;Obtain the feature point image of tissue section and DAPI staining;Find the best scaling ratio is 最佳 Of feature point image of tissue section and DAPI staining, rotation angle ia, horizontal displacement dx and vertical displacement dy and average point (xm, ym);With average point (xm, ym) generation multiple sub-frames, so that the tissue section feature point contained in sub-frame exceeds 200;For each sub-frame, using the best scaling ratio is 最佳 Rotation angle ia, in the case of (xm, ym) as rotation center, DAPI staining feature point is handled to generate transformation matrix tf2;Transformation matrix tf2 and DAPI staining image are generated final registration image by cv2.warpAffine function in opencv.Can tissue section image under normal microscope white light irradiation and DAPI staining image under fluorescence microscope are registered.