4 results about "DAPI" patented technology

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.