40 results about "Co localization" patented technology

The invention belongs to the technical field of biology, and discloses a stable protective agent, a cell antigen dry sheet and a preparation method and application of the dry sheet, the stable protective agent comprises glycerin with the volume fraction being 0.4%, gelatin with the mass volume fraction being 0.1%, EDTA with the mass volume fraction being 0.1 mM, trehalose with the mass volume fraction being 0.03% and resveratrol with the mass volume fraction being 100 [mu] M. The stable protective agent is adopted for preparing the cell antigen dry sheet, and the cell antigen dry sheet is applied to CBA detection. The stable protective agent has good permeability, can penetrate through cell membranes to enter cells, provides a protection effect inside and outside the cells, has moisture retention and oxidation resistance, can prevent cell deformation and cell fixing layer shedding, and can well maintain the stability and reliability of antigen proteins, and the prepared cell antigen dry sheet can be stored for a long time, and has a good application prospect. The antigenicity of a membrane signal index is not damaged. When a positive sample is detected, obvious green fluorescence cells can be seen, red fluorescence quenching is avoided, and co-localization of red fluorescence and green fluorescence can be observed conveniently.

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 relates to a whole genome multi-recombinant protein modification labeling method based on transposition reaction and fluorescence in situ hybridization and application thereof, and belongs to the technical field of cell molecular markers. The invention relates to a marking method modified by a whole genome multi-recombinant protein. The marking method comprises the following steps: cleaning cells carrying one or more special sequences for coding oligonucleotides; and mixing the cleaned cells, an EC buffer solution and an aqueous solution of the oligonucleotide fluorescent probe, carrying out room temperature incubation, discarding the liquid, cleaning, and carrying out fluorescence imaging. According to the labeling method, fluorescence in-situ labeling can be carried out on different types of fine histone modifications in the same cell sample, and the labeling method is suitable for an ultrahigh-resolution microscopic imaging platform or a common confocal imaging system. The marker has extremely high specificity and sensitivity, and spatial distribution and co-localization information of various histone modifications in a cell nucleus can be obtained.

This invention belongs to the field of biotechnology and discloses a dry film stabilizer, cytoplasmic antigen dry films, their preparation methods, and applications. The dry film stabilizer includes polyethylene glycol, trehalose, BSA, melatonin, and Tween-20. Cytoplasmic antigen dry films are prepared using this stabilizer and applied in CBA detection. This dry film stabilizer has good permeability, penetrating the cell membrane to enter the cell interior, providing protection both inside and outside the cell. It also possesses moisturizing and antioxidant properties, preventing cell deformation and shedding of the cell fixation layer, effectively maintaining the stability and reliability of intracellular antigen proteins. The prepared cytoplasmic antigen dry films can be stored long-term without damaging the antigenicity of cytoplasmic signal indicators. When detecting positive samples, strong and numerous green filamentous fluorescence signals are observed in the cytoplasm, with consistent green and red fluorescence intensities. Software synthesis reveals co-localization of green and red antigen fluorescence.

The invention relates to the field of medical image analysis, in particular to a drug distribution and metabolism dynamic analysis system of a multispectral microscopic image, which comprises a light source module, a drug distribution module, a drug distribution module, a drug metabolism dynamic analysis module and a drug metabolism dynamic analysis module, and is characterized in that the light source module emits a multiband light source comprising broadband white light and time sequence switching narrow-band monochromatic exciting light; the data acquisition module is used for acquiring a multi-channel spectral image signal and generating a multi-spectral image containing a spatial dimension and a spectral dimension; the pharmacokinetic analysis module constructs a geometric characterization framework of drug metabolism based on a differential geometry theory, and comprises a Riemannian manifold characterization unit for mapping multispectral data to a Riemannian manifold space, a curvature tensor analysis unit for calculating manifold curvature characteristics and a parallel transmission optimization unit for evaluating drug transmission efficiency; the application analysis module carries out drug target co-localization analysis, space-time metabolic map analysis and delivery system optimization based on the analysis result, the spectrum channel limitation of traditional fluorescence imaging is broken through, and the precision and dimension of drug metabolism analysis are remarkably improved.

The invention belongs to the field of bioinformatics and computational biology, and provides a bioinformatics multi-omics association analysis method based on a co-disease model. Comprising transcriptome range correlation analysis based on GWAS and GTEx, causal analysis in a transcriptome range based on different Bayesian models, and screening and functional analysis of co-disease hub genes based on co-localization analysis and transcriptome data. According to the method, a complete technical system from macroscopic population queue genome analysis to human body multi-tissue fine positioning and then to microscopic cellular level gene target screening and verification is established, and a reliable target gene screening process and a mechanism analysis scheme are provided for precise prevention and treatment of metabolism and vasoplasmosis.

The application discloses a kind of mitochondrial NADH detection and lipid droplet imaging bifunctional fluorescent probe and its preparation method and purposes, wherein the structure of mitochondrial NADH detection and lipid droplet imaging bifunctional fluorescent probe is as shown in the following: The bifunctional fluorescent probe of the application can produce effective optical response under the activation of NADH.The photophysical property experiment in vitro reflects that the probe can sensitively detect the level fluctuation of NADH (detection limit is as low as 12.83 nM), and confocal fluorescence microscopic imaging experiment shows that the probe has good light stability for G2 cell, and can effectively locate mitochondrion in cell (location coefficient is 0.92). With the lapse of time, the probe gradually transfers from mitochondrion to lipid droplet, and good lipid droplet localization (co-localization coefficient is 0.92) is achieved after one hour, so that fluorescence tracing of lipid droplet is realized.

The application discloses a double-color fluorescence detection anti-NMDAR antibody fixing reagent, a method and application thereof, and the fixing reagent comprises a mixture of paraformaldehyde, methanol and acetone, wherein the volume fraction of methanol in the mixture is 10-25%, and the volume fraction of acetone is 75-90%; the fixing effect is better when the fixing is carried out in two steps by using 4% paraformaldehyde in mass-volume concentration and the mixture of 15% methanol and 85% acetone in volume fraction. By using the fixing liquid and the fixing method, the antigenicity of NMDAR is not damaged, obvious green fluorescent cells can be observed in positive samples, red fluorescence is not quenched, and co-localization of red fluorescence and green fluorescence can be observed by using a fluorescence microscope.

Provided herein are methods for highly multi-plexed detection and quantification of co-localized membrane surface proteins. Provided herein are also methods for determining the likelihood that a subject has a disease or condition, such as cancer.

The invention discloses a wheat grain quality detection and functional gene analysis method based on a hyperspectral imaging technology, and belongs to the technical field of agricultural detection. The method comprises the following steps: extracting a hyperspectral index; determining a wheat grain quality index as an artificial measurement value; a weight coefficient is given to each quality index, and the wheat quality phenotype is digitalized; extracting a characteristic wavelength, and establishing a prediction model of each index and a wheat quality value; gWAS, SNP site screening and co-localization analysis are performed on the hyper-spectral index and the chemical index as phenotypic characters and genotypes respectively, and candidate genes are obtained and subjected to functional verification. The method combines a hyperspectral technology with chemometrics modeling, and is suitable for real-time monitoring in wheat quality breeding and processing processes; the spectrum phenotype is combined with GWAS gene mining, the limitation that traditional quality detection only serves phenotype screening is broken through, a'nondestructive testing-gene positioning 'closed loop is formed, and a target gene is provided for wheat quality improvement.

The application belongs to the technical field of biology, and discloses a stabilizing protective agent, a cell antigen dry sheet, and a preparation method and application of the dry sheet. The stabilizing protective agent comprises 0.4% glycerol in volume fraction, 0.1% gelatin in mass volume fraction, 0.1 mM EDTA, 0.03% trehalose in mass volume fraction, and 100 muM resveratrol. The stabilizing protective agent is used to prepare the cell antigen dry sheet and is applied in CBA detection. The stabilizing protective agent has good permeability, penetrates into the cell interior through the cell membrane, provides protection inside and outside the cell, has moisturizing and antioxidant properties, can prevent cell deformation and cell fixation layer from falling off, well maintains the stability and reliability of antigen protein, and the prepared cell antigen dry sheet can be stored for a long time without damaging the antigenicity of the membrane signal index. When a positive sample is detected, obvious green fluorescent cells can be seen, and red fluorescent quenching does not occur, so that the co-localization of red fluorescence and green fluorescence can be observed.

The invention relates to the technical field of virus detection, and discloses a multi-target fluorescent staining rapid sample preparation virus detection method, which comprises the following steps: S1, sample pretreatment; s2, multi-target fluorescent dyeing: mixing the pretreated sample in S1 with a multi-target fluorescent dye combination to obtain a dyed sample; s3, purification after dyeing; s4, optical imaging acquisition; s5, image analysis and judgment: inputting the fluorescence image in S4 into an image analysis system, extracting fluorescence characteristic parameters of each target spot, and comparing the fluorescence characteristic parameters with a preset threshold value; and S6, outputting a result. By adopting the technical scheme of combining multi-target fluorescent staining and multi-target co-localization judgment, the technical effects of accurately identifying target viruses and avoiding non-specific interference are achieved, and compared with the technical scheme of single-target fluorescent staining or single-signal judgment in the prior art, the technical scheme of the invention has the advantages that the accuracy is high; the problems that the specificity is insufficient, and false positive and false negative occur due to interference of environmental impurities or similar pathogens are solved.

The invention relates to a tetramethyl rhodamine (TAMRA)-labeled agmatine (AGM) fluorescent probe as well as a preparation method and application thereof, and relates to the technical field of biological medicines. A fluorescent group TAMRA is connected with AGM through a solid-phase synthesis method, the high-purity TAMRA-AGM probe is obtained after purification and identification, the high-purity TAMRA-AGM probe has similar biological characteristics with unmodified agmatine, and on the cellular level, the probe can effectively enter various types of intestinal epithelial cells in a short time under the condition of low dosage and is co-located on mitochondria; in a tissue level, the probe can effectively enter intestinal epithelial cells in a body and can carry out visual observation and semi-quantitative analysis on the intestinal epithelial cells; the method realizes visual tracking and accurate positioning of agmatine in cell and tissue levels, provides a key research tool for deeply illuminating the action mechanism of AGM in prevention and treatment of sepsis intestinal injury and other diseases, and has important scientific value and application prospect.

The present application relates to a kind of space co-localization's fluorescence multispectral image blind unmixing method and system.The spectral asymmetry of fluorescent protein, the spectral sparsity of fluorescent protein, the characteristics of protein kurtosis continuity are used to estimate the spectral curve of fluorescent protein, combined with the unmixing principle of minimum mutual information, fluorescence multispectral image blind unmixing is carried out, without reference prior fluorescent protein spectral curve, thereby solve the technical problem that existing unmixing method cannot carry out space co-localization's fluorescence multispectral image unmixing without fluorescent protein spectral curve.The problem to be solved by the present application is to overcome the problem that existing spectral overlapping fluorescent dye cannot be blindly unmixing when co-expressed.The spectral curve of no-reference, blind estimation is used to de-constrain existing mutual information minimization unmixing algorithm, to realize the unmixing of co-localization fluorescent protein.The present application has complete blind source separation unmixing capability, and can complete unmixing without using reference spectrum curve.

The invention discloses an intelligent slow-release drug carrier screening method based on material microscopic image recognition, which integrates nano hyperspectrum, cryoelectron microscope and Raman spectrum data through a multi-modal data synchronous acquisition system to realize nanoscale space alignment of microscopic morphology and component distribution. A non-local mean denoising algorithm is adopted to enhance pore structure recognition, and chemical bond state changes are quantified in combination with adaptive baseline correction and multi-peak fitting technologies. A characteristic spectrum library is constructed, a chemical distribution diagram is generated through spectrum angle mapping, and the co-localization of the carrier and the medicine is analyzed in combination with spatial mutual information entropy. A space-spectrum combined encoder is innovatively developed, multi-modal features are fused by using a cross attention mechanism, and quantitative indexes such as a pore filling rate, crystallinity change and an interaction stability score are output. According to the method, the static and single-mode limitation of a traditional method is broken through, and a complete screening system from microstructure analysis to release dynamics prediction is established.

The invention provides a detection method for realizing visual protein-protein interaction by co-transferring an outer nuclear membrane. The method comprises the following steps: constructing an expression vector of an outer nuclear membrane localization protein and a fluorescent protein; recombining the expression fragment of the bait protein with an expression vector to construct a bait protein expression vector; transforming and extracting plasmids; constructing a prey protein expression vector; transforming and extracting plasmids; transfecting bait protein expression vector plasmids and prey protein expression vector plasmids into cells in proportion, and culturing; and performing fluorescence imaging. And the co-localization condition of the protein on the outer nuclear membrane is directly observed by utilizing fluorescence labeling, so that the interaction of the protein can be judged. Positioning is clear and visual; and the method has high specificity, living cell compatibility and high throughput potential, so that the method has a wide application prospect in protein function research and drug screening.

The invention discloses a vacuole localization detection method based on autofluorescence of rose cyanin, and belongs to the technical field of subcellular localization. Researches find that anthocyanin in epidermal cells of rose petals has an autofluorescence characteristic and can be used as a natural marker for vacuole localization; moreover, the vacuole system of the epidermal cells of the rose petals is relatively independent, is not tightly attached to cell membranes, can be used for subcellular localization without extracting protoplast, and is a novel ideal material for researching protein vacuole localization. Based on epidermal cells of rose petals, the invention develops a novel vacuole positioning detection method, and the co-positioning condition of autofluorescence of target protein and anthocyanin is directly observed through a laser confocal microscope. The method does not need to externally add fluorescent dye, and avoids the problems of poor compatibility of markers, background interference and the like; meanwhile, a protoplast preparation step is omitted, the operation is simple and convenient, and the experimental period is short.

The invention relates to hybrid chain reactions (HCR). Specifically, the sensitivity of hybrid chain reaction (HCR) signal amplification is combined with two or more segmented initiator probes and one or more proximity probes that are capable of co-localizing a complete HCR initiator that will trigger an HCR reaction when the targets are in proximity to each other.

The invention discloses a circulating tumor cell bimodal co-localization model and application thereof. The bimodal co-localization model comprises a morphological analysis unit and an immunofluorescence signal analysis unit for circulating tumor cells, wherein the morphological analysis unit comprises Diff staining, and the immunofluorescence signal analysis unit comprises a CD45 mouse antibody, a 488-mouse secondary antibody, a CK rabbit antibody and a 594-rabbit secondary antibody. The circulating tumor cell bimodal co-localization model has the advantages that in the application process of the circulating tumor cell bimodal co-localization model, a non-antibody-dependent nano magnetic bead enrichment technology is adopted, so that the completeness and cell viability of CTC are improved, and the success rate of subsequent multi-step dyeing is ensured; the use amount of the sample and the collection frequency are reduced; the use amount of the peripheral blood sample is reduced through double analysis (morphology and molecular markers), so that the clinical universality is improved; in combination with morphology and immunofluorescence detection, the risk of false positive and false negative is reduced, and the diagnosis accuracy of CTC is improved.

The invention relates to a method for detecting the state of a DNA nanostructure based on co-localization of a single dye label and a subcellular organelle, and the detection method comprises the following steps: incubating a DNA nanostructure connected with a dye and cells, and detecting the co-localization coefficient of the dye and the cellular organelle; when the co-localization coefficient of the dye and the lysosome is greater than 0.5, judging that the DNA nanostructure is complete; when the co-localization coefficient of the dye and the mitochondria is greater than 0.5, judging that the DNA nano structure is damaged; and judging that the DNA nanostructure is partially complete under other conditions. According to the method for judging the fate of the DNA nanostructure and detecting the integrity of the DNA nanostructure based on co-localization of the single dye and the subcellular organelle, not only can the intact or damaged states of the same DNA nanostructure under different conditions be distinguished by utilizing the position of the DNA nanostructure in the subcellular organelle in the cell, but also the intracellular states of different DNA nanostructures can be detected.

The invention provides a tumor cell mitochondria targeting agent with a fluorescence co-localization function and a preparation method thereof, and belongs to the field of biomedicine. According to the method, triphenylamine is taken as a raw material, a pi conjugated skeleton is expanded by a benzothiadiazole bridging unit, pyridinium salt is taken as a strong electron acceptor, and different alkyl chains are connected, so that the mitochondrial targeting agent with aggregation-induced emission property is synthesized. The fluorescent mitochondrial targeting agent TBPMI not only has good biological safety, but also has good tumor cell mitochondrial selectivity and targeting accuracy, and the co-localization coefficient of the fluorescent mitochondrial targeting agent TBPMI and a commercial mitochondrial probe is as high as 0.96. Meanwhile, the prepared fluorescent mitochondrial targeting agent TBPMI has excellent aggregation-induced emission performance, and the aggregation-induced emission performance is obviously higher than that of a traditional commercial photosensitizer rose bengal. Therefore, the newly prepared fluorescent mitochondrial targeting agent TBPMI provides a new material for accurate positioning of organelle level under the guidance of a fluorescent image.

This application relates to the field of coronal mass ejection (CME) detection technology, and discloses a method, system, device, and medium for identifying CMEs. The method for identifying CMEs includes: acquiring a coronal observation image; learning a global feature map of the coronal observation image using a self-supervised learning method; learning a hierarchical feature representation of the global feature map through a cascaded multilayer PCA filter to obtain a co-localization map of CMEs, wherein the co-localization map represents a potential distribution area of ​​CMEs; obtaining a minimized global energy function, and performing fine segmentation of the potential distribution area of ​​CMEs using the minimized global energy function to obtain the region segmentation result of the CMEs. Using this application, the efficiency and accuracy of coronal mass ejection detection are effectively improved.

The application discloses a molecular recruitment co-localization system, comprising: a spatially targeted visualization module for recruiting target proteins to a specific site and generating a fluorescence co-localization signal in living cells based on the target proteins recruited to the specific site; a multi-protein interaction verification module for observing protein interactions in a multi-protein complex based on different pairs of proteins recruited to the specific site at a specified genetic locus; a protein binding capacity evaluation module for quantitatively analyzing the binding capacity between proteins based on the co-localization percentage and signal-to-noise ratio of the proteins; a protein phase separation capacity evaluation module for rapidly evaluating whether different proteins or protein domains have phase separation capacity based on the recruitment of the different proteins or protein domains to a specific location; and a dynamic observation and quantitative evaluation module for observing the dynamic changes of proteins in the phase separation process in real time by combining multi-channel fluorescence and quantum dot probes and providing preliminary quantitative data for the physical properties of the condensate.

The invention discloses a multiple immunofluorescence detection method and equipment based on spatial reconstruction and a medium, and relates to the technical field of histopathology analys.The multiple immunofluorescence detection method comprises the steps that a tissue sample is prepared, fluorescent reference microbeads are added, and continuous slices are obtained; circularly preprocessing the tissue sample, and performing intensity normalization and bleaching time drift correction by using fluorescent reference beads to obtain a corrected image sequence; performing spectral unmixing under the three-dimensional voxel coordinate system, and introducing spatial regular constraint to obtain a voxel-level fluorescence intensity vector; reconstructing a high-resolution voxel stack and segmenting a cell voxel set by combining a nuclear dye signal and a fluorescent reference microbead and adopting rigid and non-rigid registration and assisted by three-dimensional deconvolution; and calculating a proximity relation and a co-localization index of different phenotype cells, and generating a detection result. According to the invention, high-resolution tissue and cell reconstruction in a three-dimensional space is realized, the space continuity and accuracy are improved, and the stability and accuracy of voxel-level fluorescence signal quantification are improved.

The application discloses an intelligent sustained-release drug carrier screening method based on material microscopic image recognition, integrates nanometer hyperspectral, cryo-EM and Raman spectral data through a multi-modal data synchronous acquisition system, realizes nanoscale spatial alignment of microscopic morphology and component distribution, adopts a non-local mean denoising algorithm to enhance pore structure identification, combines adaptive baseline correction and multi-peak fitting technology, and quantifies chemical bond state changes, constructs a characteristic spectral library, generates a chemical distribution map through spectral angle mapping, combines spatial mutual information entropy to analyze the co-localization of carriers and drugs, innovatively develops a space-spectrum joint encoder, fuses multi-modal features by using a cross attention mechanism, and outputs quantitative indexes such as pore filling rate, crystallinity change and interaction stability score. The method breaks through the static and single-modal limitation of traditional methods, and establishes a complete screening system from microscopic structure analysis to release kinetics prediction.

Despite the advance of screening technology, omic-based studies with spatial resolution still requires laborious efforts, hampering the analysis of biology and disease. The present disclosure provides methods, systems, probes, and platforms that may be based on the use of flow-based sequencing to increase the throughput of analyte screening with spatial resolution.

The invention discloses a molecular recruitment co-localization system, comprising: a space fixed-point visualization module for collecting a target protein to a specific site and generating a fluorescent co-localization signal in a living cell based on the target protein collected to the specific site; a multi-protein interaction verification module for observing protein interaction in a multi-protein complex based on recruiting pairs of different proteins recruited to a specific site on a specified locus; the protein binding capacity evaluation module is used for quantitatively analyzing the binding capacity between the proteins based on the co-localization percentage and the signal-to-noise ratio of the proteins; the protein phase separation capability evaluation module is used for rapidly evaluating whether different proteins or protein structural domains have phase separation capability or not on the basis of recruiting the different proteins or protein structural domains to specific positions; and the dynamic observation and quantitative evaluation module is used for observing the dynamic change of the protein in the phase separation process in real time by combining multi-channel fluorescence and a quantum dot probe, and providing preliminary quantitative data for the physical characteristics of the aggregate.

The invention discloses a monoamine oxidase inhibitor type near-infrared fluorescent probe, a synthesis method thereof and application of the monoamine oxidase inhibitor type near-infrared fluorescent probe in photothermal therapy of tumors. The method comprises the following steps: (1) synthesizing the monoamine oxidase inhibitor type near-infrared fluorescent probe SINH; and (2) applying the SINH probe to monoamine oxidase activity inhibition, organelle co-localization imaging and targeted phototherapy tumor treatment. The problem that a traditional inhibitor type probe only targets tumors and does not combine with illumination for phototherapy is solved; the inhibitor type near-infrared fluorescent probe has near-red absorption and emission properties, so that interference of background fluorescence signals can be weakened, and the tissue penetration depth can be increased; the probe disclosed by the invention not only has strong inhibition on the growth of SH-SY5Y cells with high MAO expression, but also can be used for carrying out photodynamic and photothermal therapy on subcutaneous tumor-bearing mice with high MAO expression, and has a very good inhibition effect on the growth of tumors with high MAO expression.

The present invention belongs to the technical field of single-cell chemical imaging, and specifically is a stimulated Raman microscopy imaging device combined with optical tweezers. The device of the present invention includes a laser optical tweezers optical path, a Raman imaging optical path, and a microscope; the optical tweezers optical path couples the laser to the microscope objective through a dichroic mirror to achieve strong focusing and form a strong gradient force optical trap; the Raman optical path contains two pulsed lights that are synchronized in time domain and overlap in space. The two pulsed lasers are scanned by a scanning galvanometer and then coupled to the microscope by the optical path system to achieve two-dimensional scanning of the light beam on the sample; the optical tweezers use a position-sensitive detector to achieve real-time recording of the position of samples such as cells, and Raman imaging uses a photomultiplier tube or a photodetector after phase-locked amplification and output. The co-localized Raman light source can achieve chemical imaging of multiple components of the sample by scanning. The invented device can be used to perform Raman co-localization imaging of multiple chemical components in cells, and has important applications in the fields of cell component analysis and component distribution measurement.