18 results about "Colocalization" patented technology

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 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 present invention belongs to the technical field of sensorineural hearing. Provided is the use of SPTBN1 as a target in the treatment of GJB2-related sensorineural hearing loss. The use comprises: the construction of a stably transfected cell line, wherein: HEK293T cells are cultured using a DMEM culture medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin in a humidified incubator at 37ºC with 95% air and 5% CO2, and when the cells reach 80%-90% confluence, cell passage is performed; IP-MS analysis, wherein: protein complexes are purified using Protein A / G immunoprecipitation magnetic beads, a portion of the extracted proteins is used as input, and then 2 μg of anti-Cx26 antibody is added to the remaining protein extract, same are gently pipetted and mixed, and incubated on a rotating shaker at 4℃ overnight; immunofluorescence observation of the co-localization of Cx26 and SPTBN1, wherein: a stably transfected cell line expressing WT-Cx26 and Mut-Cx26 is constructed; and co-immunoprecipitation (Co-IP) validation, wherein: a stably transfected cell line expressing WT-Cx26 is constructed. The present invention overcomes the limitations of therapeutic methods, such as the relatively low targeting specificity and short therapeutic time windows associated with full-length protein supplementation via gene therapy, thereby providing new insight into GJB2-related hearing loss, and a new target and a new treatment for same.

The application discloses a fluorescence enhancement anchor agent based on a fluoroboron dipyrrin-tetrazine dye for inflation microscopy technology, and belongs to the technical field of biochemical analysis. The fluorophore of the fluorescence enhancement anchor agent is shown in formula (I), formula (II) or formula (III): the tetrazine group carried by the fluorescence enhancement anchor agent is removed from fluorescence quenching through a biological orthogonal reaction with a drug probe, and the drug probe is shown in formula (IV): the fluorescence enhancement anchor agent can covalently and non-covalently anchor small molecules in a hydrogel network based on a tetrazine biological orthogonal reaction, and realizes inflation microscopic fluorescence imaging of a double fluorescence enhancement mechanism. In addition, the drug probe selects a non-covalent drug, gefitinib, as a probe core, is modified with a BCN click handle, and successfully realizes in-situ fluorescence inflation microscopic imaging of an EGFR drug-target, thereby providing a new tool for research and clinical research on non-covalent drug target co-localization information of exosomes.

This invention relates to the field of biomedical image processing technology, and discloses a method and system for quantitative analysis of hypothalamic immunofluorescence images. The method includes: performing spectral unmixing on multispectral fluorescence microscopy images based on a nonnegative matrix factorization algorithm to obtain a clean signal distribution map; using Gaussian Laplace filtering and watershed transform to achieve cell detection and segmentation; performing affine and B-spline registration between slice images and standard brain atlases to generate regions of interest masks for neural nuclei; using a local background adaptive correction strategy to perform fluorescence quantification and positive determination; and calculating Pearson correlation coefficient and Manders overlap coefficient to achieve colocalization analysis. The system includes a spectral unmixing module, a cell detection and segmentation module, an atlas registration and region recognition module, a fluorescence intensity quantification module, and a colocalization analysis and statistical output module.

The invention discloses an embryo development quality evaluation method and device, and belongs to the field of embryo in-vitro culture, the embryo before implantation is subjected to three-dimensional imaging to form a three-dimensional blastocyst structure chart, fluorescence imaging is carried out to form a redox specific heat chart, the three-dimensional blastocyst structure chart and the redox specific heat chart are subjected to co-localization superposition, and the embryo development quality is evaluated. Constructing a three-dimensional blastocyst model based on map combination; according to the three-dimensional blastocyst model, the spatial form, chromosome ploidy and metabolic state of the early blastocyst are comprehensively evaluated, and non-intrusive, accurate and efficient blastocyst ectodevelopment quality evaluation and transplantation screening are achieved.

The application relates to a mitochondrial membrane potential detection probe technology, in particular to a dual-color fluorescence probe for self-referenced monitoring of mitochondrial membrane potential and application thereof.The probe comprises a compound CBTH or pharmaceutically acceptable salts, hydrates, solvates, optical isomers, racemates and the like of the compound CBTH.The chemical structural formula of the compound CBTH is shown as formula (I).The probe CBTH can be imaged simultaneously in dual colors, the fluorescence intensity does not disappear with the change of the cell state, and the change of the mitochondrial membrane potential can be monitored semi-quantitatively by calculating the colocalization coefficient.The probe has low toxicity, good biocompatibility, reduces the cumbersome operation of co-staining of two probes, and is expected to be widely applied in the research on diseases and physiological processes related to mitochondria.

The invention relates to the technical field of biological detection, in particular to a method for detecting luminous efficiency of fluorescent protein based on double-color co-localization, which comprises the following steps: providing a double-labeled probe which is a fusion protein and comprises a self-labeled label, transmembrane protein or a transmembrane fragment thereof and target fluorescent protein which are connected in sequence; the self-labeling label releases a first detectable signal after being combined with a substrate, the target fluorescent protein releases a second detectable signal, and the first detectable signal is different from the second detectable signal; enabling host cells to express the double-labeled probe, and detecting the first detectable signal and the second detectable signal, so as to analyze and obtain the luminous efficiency of the target fluorescent protein. The method can be used for determining the absolute luminous efficiency of various fluorescent proteins, and has the advantages of simplicity and convenience in operation, high accuracy and wide application range.

A method for high dimensional spatial analysis includes segmenting, into a plurality of segments, an image depicting a plurality of cells comprising a biological sample. Each segment of the plurality of segments may correspond to one cell of the plurality of cells. A phenotype for each cell of the plurality of cells depicted in the image may be determined based on the segmented image. The determining of the phenotype may include identifying, within the plurality of cells, a first cell type having a first phenotype and a second cell type having a second phenotype. One or more metrics, such as a colocation quotient or an Earth Mover's Distance, quantifying a co-occurrence pattern between the first cell type and the second cell type may be determined. A visual representation of the co-occurrence pattern between the first cell type and the second cell type may be generated based on the metric.

The invention discloses a mitochondrial targeting near-infrared fluorescent probe and a preparation method and application thereof, and belongs to the field of medical living body fluorescence imaging, and the mitochondrial targeting near-infrared fluorescent probe designed and synthesized by the invention has high sensitivity (18.6 nM), quick response (5 minutes) and mitochondrial targeting characteristics (co-localization coefficient is 0.925). And an ideal tool is provided for dynamically monitoring the mitochondrial oxidation stress state in the hypoxic pulmonary hypertension model.

The invention belongs to the field of biomedical materials, and particularly relates to a nuclear magnetic fluorescence bimodal imaging probe and a preparation method and application thereof. The invention provides a nuclear magnetic fluorescent bimodal imaging probe which comprises superparamagnetic iron oxide particles, near-infrared fluorescent dye, a targeting ligand and a biocompatible polymer, and the superparamagnetic iron oxide particles, the near-infrared fluorescent dye and the targeting ligand are covalently coupled through the biocompatible polymer. And a polyphenol compound is used as a link to form metal-ligand coordination. The probe endows the material with stronger mechanical properties and stability through a covalent cross-linking and coordination cross-linking dual-network structure, so that the probe is not easy to depolymerize after entering a human body, it is ensured that the superparamagnetic iron oxide particles and the near-infrared fluorescent dye can be co-localized at a tumor, and the imaging accuracy is improved.

The present invention addresses the problem of providing: a nerve cell analysis method with which the localized amount of TDP-43 in nerve cells can be analyzed without labeling TDP-43 with a fluorescent tag or the like and without carrying out gene transfer; a kit for carrying out the nerve cell analysis method; and a screening method for a prophylactic and / or therapeutic agent for a neurodegenerative disease. The present invention provides a nerve cell analysis method involving: a staining step for immunofluorescent staining of nerve cells using an anti-TDP-43 antibody and an antibody that recognizes a stress granule marker; a cell region identification step for identifying the cytoplasm and nuclei of the nerve cells; and an analysis step for analyzing a TDP-43-derived fluorescence signal and a stress granule marker-derived fluorescence signal in the cytoplasm, and analyzing the localized amount of TDP-43 in the nerve cells on the basis of the presence or absence of colocalization of a granular TDP-43 signal and a granular stress granule marker signal in the cytoplasm.

The invention relates to a mitochondrial membrane potential detection probe technology, in particular to a double-color fluorescent probe for self-reference monitoring of mitochondrial membrane potential and application of the double-color fluorescent probe. Comprising a compound CBTH or pharmaceutically acceptable salts, hydrates, solvates, optical isomers, racemes and the like. The chemical structural formula of the compound CBTH is as shown in formula (I): # imgabs0 #. The probe CBTH disclosed by the invention can be used for simultaneously imaging two colors, the fluorescence intensity does not disappear along with the change of the cell state, and the change of the mitochondrial membrane potential can be semi-quantitatively monitored by calculating a co-localization coefficient. The probe is low in toxicity and good in biocompatibility, the tedious operation of co-dyeing of the two probes is reduced, and the probe is expected to be widely applied to the research of mitochondrial related diseases and physiological processes.

The invention discloses boron-rich polymer nanoparticles with organelle targeting as well as a preparation method and application of the boron-rich polymer nanoparticles. The naphthalimide and the dodecahydrododecaborate are subjected to azide-alkyne cycloaddition to form the boron-containing medicine. The boron-rich polymer nanoparticle is formed by electrostatic self-assembly of a boron-containing drug and a block polymer, has the particle size of 20-200nm, has a pomegranate-like structure, and has the functions of high boron content (greater than or equal to 10wt%), low dark toxicity (the cell survival rate is greater than or equal to 90%), good biocompatibility, specific tumor specific organelle targeting (the co-localization coefficient is greater than or equal to 0.82) and fluorescence imaging tracing. The method can be used for monitoring the treatment process in real time. According to the invention, the effect of boron neutron capture therapy (BNCT) is obviously improved, side effects on normal tissues are reduced, and the method has a wide application prospect in the field of tumor treatment.

The application belongs to a kind of difluoroborane hydrazine pyrrole fluorescent probe and its preparation method and beta amyloid detection application, belongs to the field of fluorescence detection technology. The probe is composed of BOPHY fluorescent mother body and recognition group, and its preparation method includes three steps of BOPHY precursor preparation, fluorescent mother body synthesis and target probe molecule synthesis. The prepared series of probes have large Stokes shift, can effectively reduce imaging interference, and show high sensitivity and high selectivity of fluorescence response to A beta aggregate, wherein the performance of N-BOPHY is optimal. Experiments show that the binding affinity of the probe to A beta aggregate is high, the staining imaging result of A beta plaque in brain slice has high colocalization with commercial probe thioflavin-T, and the cell toxicity is low and the biocompatibility is good.

A multimodal optical technique that can measure the mechanical, optical, and acoustical properties of the sample at microscopic resolution, which is based on the integration of a Brillouin microscope and a photoacoustic (PA) microscopy is provided. The multimodal technique not only can acquire co-registered Brillouin and PA signals of the sample but also allows us to utilize the sound speed measurements by PA to quantify the sample's refractive index, which is an essential property of the material and cannot be measured by either technique individually. We demonstrated the colocalization of Brillouin and time-resolved PA signals by measuring the interface of kerosene and 1% CuSO4 aqueous solution. In addition, we measured the refractive index of saline solutions with a precision of 0.003 and validated the result against published data. This multimodal modality could open new way for charactering biological cell and tissue in physiological and pathological conditions.