38 results about "Cell sorting" patented technology

The invention discloses a method for separating, culturing and identifying porcine muscle-derived mesenchymal stem cells and fat progenitor cells. The method mainly comprises the following steps: dissociating pig muscle tissues through a tissue processor, obtaining mesenchymal stem cells and adipose progenitor cells by utilizing a flow cytometry sorting technology, and completing in-vitro culture and identification of the cells. According to the technical scheme provided by the invention, the technical problems of primary culture, separation and identification of the mesenchymal stem cells and the adipose progenitor cells of the porcine muscle tissue are effectively solved, and a reliable method is provided for further application of a flow cytometry sorting technology and a myogenic mesenchymal stem cell and adipose progenitor cell primary model in animal nutrition research.

The present application relates to the technical field of microfluidic chip, and specifically discloses a microstructure spiral inertia microfluidic chip for cell deformation ability sorting and application thereof, the chip comprises a spiral flow channel, an inlet is arranged at the upstream end, a first outlet and a second outlet are arranged at the downstream end, and micro-pillars are arranged on the inner side wall of the spiral flow channel towards the curvature center at equal intervals. The present application strengthens the Dean secondary flow through the micro-pillar structure, and in a wide flow rate range, cells with different deformation abilities are respectively focused on the near inner wall and far inner wall regions of the flow channel based on the force balance difference, and high-throughput sorting without barriers is realized through the corresponding outlets, the sorting flux is more than 2 million cells / min, the migration and invasion ability of the soft cancer cell subpopulation obtained by sorting is 2-3 times that of hard cells, high-speed imaging can be matched to realize online quality control, and technical support is provided for tumor mechanical phenotype research.

In one embodiment, a system includes a flow cell with a conductive electrode and a base having an opening allowing variable charged sheathed sample fluid to flow out along a fluid axis, a droplet deflection unit with a pair of charge plates and a deflection chamber with a pivot door closing over the chamber a hardware triggered camera mounted to the back of the deflection chamber, and an LED array strobe light mounted to the pivotal door opposite the camera. The LED array strobe light generates a backlighting for various droplet streams. Triggering of the camera is synchronized with the generation of the backlighting to periodically capture a brightfield still image of deflected droplets and center droplets in the deflection chamber. The captured images can be analyzed for droplet deflection angles and center line for visual feedback control of the deflected droplets and the center droplets in respective droplet streams.

The utility model discloses a cell sorter roll-over stand which comprises a fixed frame, the fixed frame is fixedly or detachably installed on an operation base station, a cell sorter supporting frame and a material receiving groove are arranged on the fixed frame, the fixed frame is of a semi-surrounding structure, and the material receiving groove is arranged on the fixed frame. The cell sorter supporting frame is arranged on the fixing frame and connected with the fixing frame in a hinged mode, the material receiving groove is connected with the fixing frame in a hinged mode, a cell sorter is arranged on the cell sorter supporting frame, and a test tube fixing opening used for installing a test tube is formed in the material receiving groove. The cell sorting device has the advantages that the structure is simple, the operation time of experimenters is saved, the overall working efficiency is improved, and in the use state, the poured liquid is poured into the test tubes to be directly collected, so that the operation is more efficient and convenient.

The present application provides a kind of image vignetting correction method, image processing device and application, this method is under the condition of predetermined photographing environment, using camera to shoot template image under the condition of welding and carry out gray value sampling, surface fitting, with the maximum value of fitting surface equation divided by the surface fitting value of template image, and the result is normalized to obtain normalized dark corner compensation model, based on the normalized dark corner compensation model, the image to be corrected that camera shoots under the condition of predetermined photographing environment is corrected.The present application can quickly find the dark corner compensation model based on template image using simple surface fitting method, correct the image to be corrected, improve image quality, without obtaining complex geometric parameters of optical system in advance, has wide adaptability.The image processing device of the present application can be applied to droplet digital PCR system to improve the accuracy of PCR detection results, and can also be applied to single cell sorting system to facilitate the identification and positioning of nozzle, improve the efficiency of single cell sorting.

Provided are a process for preparing genetically modified T cells by using PBMCs as a raw material without T cell sorting, cells produced by means of the process and the use thereof. The provided process for preparing modified T cells can reduce costs of materials, manpower consumption, and the use amount of viruses.

The application discloses a method for in-situ collection and statistics of marine cyanobacterial symbiotic system and free cyanobacteria, which comprises the following steps: firstly, multiple gradient filtration pretreatment is performed on seawater samples, and the samples are sequentially filtered through 5 μm, 3 μm (including secondary filtration and slight ultrasonic) and 0.22 μm filter membranes, so that the samples are preliminarily separated according to particle sizes and different samples are obtained, and false symbiotic systems are removed; then, flow cytometry sorting is performed, eukaryotic algae interference is removed according to the differences of five light signals of FSC, SSC, APC, PE and PerCP-cy5.5, and symbiotic cyanobacteria, symbiotic heterotrophic bacteria and free cyanobacteria are sorted out; and the sorting results can be verified through 16S amplicon sequencing. The application does not intervene in biochemistry in the whole process, can keep the original state and natural proportion of microorganisms to the maximum extent, and realizes in-situ, accurate separation and statistics of marine cyanobacterial symbiotic system and free cyanobacteria.

In one embodiment, a system includes a flow cell with a conductive electrode and a base having an opening allowing variable charged sheathed sample fluid to flow out along a fluid axis, a droplet deflection unit with a pair of charge plates and a deflection chamber with a pivot door closing over the chamber a hardware triggered camera mounted to the back of the deflection chamber, and an LED array strobe light mounted to the pivotal door opposite the camera. The LED array strobe light generates a backlighting for various droplet streams. Triggering of the camera is synchronized with the generation of the backlighting to periodically capture a brightfield still image of deflected droplets and center droplets in the deflection chamber. The captured images can be analyzed for droplet deflection angles and center line for visual feedback control of the deflected droplets and the center droplets in respective droplet streams.

The application relates to a bait vector in the field of biotechnology, a construction vector containing the bait vector, a construction method of a double-targeting RNA lentivirus library and the double-targeting RNA lentivirus library. The construction method comprises the following steps: mixing a double-targeting RNA co-expression lentivirus vector and a bait vector to prepare a vector mixture; an insertion segment Y located between 5' LTR and 3' LTR of the double-targeting RNA co-expression lentivirus vector comprises an independent targeting RNA1 expression box, a targeting RNA2 expression box and a complete reporter gene; the bait vector and the double-targeting RNA co-expression lentivirus vector exist in packaging competition and do not have functional activity of generating a complete reporter gene phenotype; a packaging cell is co-transfected with the vector mixture and a lentivirus packaging plasmid, virus packaging is carried out, viruses are collected, target cells are infected, a cell group with positive reporter gene expression is sorted out, and a double-targeting RNA lentivirus library is constructed. The method can effectively reduce the recombination rate.

A collecting system with a magnetically coupled sample mover is provided for flow cytometry and cell sorter systems. The collecting system uses magnets in a driver carriage to control the position of other magnets in a follower carriage. The driver carriage can thereby control the position of the follower carriage without physically touching the follower carriage.

The invention relates to the technical field of single-cell sequencing, and discloses a high-quality single-cell Hi-C library building method, which comprises the following steps: by utilizing a cumulative effect, carrying out cross-linking immobilization, enzyme digestion, biotinylated nucleotide filling-in and connection on population cells, sorting single cells, immediately marking different bar code combinations on each cell, mixing extracts of each cell, and carrying out high-quality single-cell Hi-C library building. After mixing, the cell population is equivalent to a small amount of cell population, enriching biotinylated DNA by using streptavidin magnetic beads, and finally establishing a library. According to the method, the library quality can be remarkably improved while the single cell resolution is maintained.

This invention discloses a method for microscopically separating and rescuing cells from heavily contaminated cell culture media, belonging to the field of biotechnology. This invention combines flow cytometry sorting technology with penicillin-streptomycin for the separation, rescue, and culture of healthy cells. The method provided by this invention can effectively rescue heavily contaminated cells, remove their contamination state, and make them usable again. This will greatly facilitate cell preservation, passage, and culture, effectively avoid the waste of cell resources, and improve the preservation rate of rare cell resources, providing strong support for the smooth progress of scientific research.

A circular nozzle assembly is disclosed. A cuvette nozzle subsystem (assembly) for flow cytometry systems is disclosed including a cuvette assembly and a circular nozzle assembly selectively engaged with the cuvette assembly. The cuvette assembly includes a cuvette having a pocket and a flow channel, and a receptacle coupled within the pocket to the cuvette. The receptacle has a through-hole with a tapered conical portion and a circular cylindrical portion. The circular nozzle assembly includes an o-ring gasket coupled to a nozzle body with a flow channel. A tapered conical portion of the nozzle body engages the tapered conical portion of the through-hole to align the respective flow channels of the cuvette and the nozzle body together.

This invention discloses a method for characterizing and isolating gut microbiota capable of specifically uptake or metabolizing natural products, comprising: in vitro co-culture, click chemistry reaction, and flow cytometry sorting. Based on click chemistry technology, this invention co-cultures alkyne-modified natural products with fecal microbiota. Through a copper-catalyzed azido-alkynyl cycloaddition reaction, azidolated fluorescent dyes are labeled onto all bacteria capable of uptake and utilizing natural products or their metabolites. These bacteria are then isolated using flow cytometry sorting. Compared to methods for studying metabolism and utilization using single-species culture, this invention is lower in cost and shorter in time, providing a new approach for extensive and in-depth research on the interaction between gut microbiota and natural products.

A method for inducing and activating a conventional dendritic cell subset, the method comprising the following steps: A. seeding human peripheral blood stem cells or umbilical cord blood stem cells in a plate, and performing expansion culture using a basal medium supplemented with cytokine 1; B. performing in vitro induced differentiation culture on the cells after the expansion culture using a basal medium supplemented with cytokine 2; C. sorting the differentiated cells to obtain pure cDC1 cells; and D. performing activation culture on the pure cDC1 cells using a basal medium supplemented with a stimulator to obtain activated cDC1 cells. A large number of human primary cDC1 are obtained by means of induction from umbilical cord blood stem cells, and the anti-tumor ability of cDC1 is then activated by means of a combination of stimulators. Moreover, the feasibility and efficacy of cDC1 for tumor therapy are verified for the first time by means of a humanized mouse tumor model.

The application relates to the field of biological medicine, and particularly discloses a monoclonal antibody combined with human CD8 and a preparation method and application thereof. The antibody is obtained by combining single B cell sorting, gene cloning and mammalian cell expression technology after rabbit immunization of a CD8 polypeptide antigen, a New Zealand white rabbit is immunized by adopting a KLH coupled CD8 antigen polypeptide combined with a rapid immunoadjuvant, high-affinity antibody clones are obtained by single B cell sorting technology, and the antibody is prepared by recombination expression. The antibody is verified by ELISA and flow cytometry, has high titer and strong specificity, can effectively recognize human peripheral blood CD8+ T cells, and has a signal strength and a background signal-to-noise ratio which are significantly better than those of a traditional mouse-derived antibody. The application provides light and heavy chain variable region amino acid sequences of the antibody, realizes controllable preparation of the antibody and protection of intellectual property rights. The antibody can be widely applied to the fields of flow cytometry detection, immunohistochemistry, tumor immunotherapy monitoring and T cell function research, and has important scientific research and clinical application values.

The invention discloses a method and device for sorting PBMC subtypes through magnetic beads, and belongs to the technical field of cell sorting. The method comprises the steps that PBMC is obtained through density gradient centrifugation; jointly incubating the PBMC, a biotinylation lineage antibody cocktail and a DCs fluorescent antibody, and then adding streptavidin coupled magnetic beads to label non-target cells; adding the cell suspension into a low-temperature magnetic sorting device, adsorbing and removing non-target cells in a magnetic field of 0-4 DEG C, and directly collecting a DCs pre-enriched suspension; and the DCs of HLA-DR + Lin-are sorted out through flow cytometry. The device comprises a sorting pipe, a magnetic shell and an ice storage bin, and can maintain a low-temperature environment and realize centrifugation-free operation. According to the method, a pedigree negative removal strategy is adopted, advanced activation of DCs is avoided, two-stage sorting and a special low-temperature device are combined, the problem that efficiency, purity and activity of DCs sorting are difficult to consider at the same time is effectively solved, and high-purity and high-activity dendritic cells can be obtained.

The invention discloses an anti-helicobacter pylori monoclonal antibody as well as preparation and application thereof, and belongs to the technical field of medical detection. The antibody is prepared by taking inactivated helicobacter pylori ATCC 43504 whole bacteria as an immunogen, immunizing a New Zealand white rabbit and then carrying out single B cell sorting, limited dilution, gene cloning, plasmid construction and eukaryotic cell expression, has a specific CDR core sequence, is high in specificity and affinity, can accurately recognize a helicobacter pylori specific antigen, and can be used for preparing a specific antibody. The cross reaction with other bacteria or tissue impurities is effectively avoided. The kit can be used for detecting the helicobacter pylori, and a new tool and method are provided for pathological diagnosis of helicobacter pylori infection and related digestive system diseases.

The invention provides an anti-human CD8 rabbit recombinant monoclonal antibody and application thereof. The antibody comprises a heavy chain variable region VH and a light chain variable region VL, a rabbit is immunized with a KLH-coupled CD8 polypeptide antigen, spleen lymphocytes are separated after titer detection is qualified, and single B cell sorting is carried out through a biotin-labeled antigen; positive clone is obtained by a rabbit single B cell culture technology, RNA (Ribonucleic Acid) is extracted and is reversely transcribed into cDNA (Complementary Deoxyribonucleic Acid), and light and heavy chain variable region genes are amplified and cloned into an expression vector for recombinant expression; and finally, the rabbit recombinant monoclonal antibody with high affinity is obtained. The antibody has the capacity of specifically recognizing CD8 + cells in human peripheral blood, is high in affinity and low in animal origin, is suitable for immunodetection such as flow cytometry and ELISA (enzyme-linked immunosorbent assay), and can be used in the fields of immune cell typing, immune function evaluation, immunotherapy monitoring and the like. The invention also provides a light and heavy chain variable region amino acid sequence of the antibody, and a basis is provided for intellectual property protection and further development of the antibody.

The application relates to the technical field of biological medicine, and particularly discloses a monoclonal antibody combined with human CD4 and a preparation method and application thereof. The antibody is obtained by combining single B cell sorting, gene cloning and mammalian cell expression technology after rabbit immunization of a CD4 polypeptide antigen. The antibody has high affinity and good specificity, and can effectively recognize CD4 positive T cells in human peripheral blood. The antibody is verified by ELISA and flow cytometry, and has high titer, low background and a signal-to-noise ratio superior to that of existing mouse-derived antibodies. Further, the amino acid sequences of the light chain and the heavy chain variable region of the antibody are obtained, so that controllability and repeatability of antibody expression are realized. The antibody can be widely applied to scenes such as immunodetection, HIV auxiliary diagnosis, T cell subgroup research and antibody engineering development, and has important scientific research and industrial values.

The invention discloses a construction method of an IER2-T2A-EGFP gene knock-in mouse model and application of the IER2-T2A-EGFP gene knock-in mouse model in myeloid cell sorting, and the method comprises the following steps: aiming at a knock-in target of a mouse IER2 gene, designing sgRNA with a sequence as shown in SEQ ID No: 2; the constructed PX459-sgRNA recombinant plasmid is used as a template, and a single-chain sgRNA + tracr RNA is obtained through PCR (Polymerase Chain Reaction) amplification and in-vitro transcription; synthesizing a Donor sequence containing the T2A-EGFP gene in an overlapping PCR (Polymerase Chain Reaction) mode; a single-chain sgRNA + tracr RNA, a Donor sequence and Cas9 protein are co-injected into male pronucleus of a mouse fertilized egg, and a female mouse is transplanted for inoculation, so that the F0-generation IER2-T2A-EGFP gene knock-in mouse model is obtained. According to the method, the primary myeloid cells can be quickly, conveniently and accurately sorted.

The present application relates to the field of immunodetection technology, and particularly relates to a method for enriching and detecting antigen-specific T cells. The method for enriching antigen-specific T cells comprises the following steps: stimulating a whole blood sample with a specific antigen, and then sorting T cells in the whole blood sample stimulated by the antigen by using an immunomagnetic bead sorting technology to obtain a T cell sample. The method is based on the strategy of stimulating the antigen first and then sorting the cells, and can quickly and accurately separate T cells from a complex cell system of whole blood, effectively avoiding the complicated operation of traditional sorting methods, realizing automatic sorting, simplifying the operation steps, shortening the sorting time, improving the sorting throughput, and better ensuring the accuracy of subsequent cell detection. The T cells obtained by the above enrichment method can be detected by using an ELISPOT method, and the antigen-specific T cells can be accurately detected.

The invention relates to a protein interaction screening system based on a bacterial two-hybrid technology, which comprises a bait protein and prey protein co-expression plasmid pCDFduet1-lambda cI-RNAP, the sequence of which is as shown in SEQ ID No. 1; a reporter plasmid is pACYC184-cIamp; the sequence of the lac promoter-GFP is as shown in SEQ ID No. 2, and the lac promoter-GFP is as shown in SEQ ID No. 2; the positive control plasmid is a pCDFduet1-lambda cI-LG2-RNAP-Gal11p plasmid, and the sequence of the positive control plasmid is as shown in SEQ ID No. 3; the invention further relates to a high-throughput protein interaction screening method of mycobacterium tuberculosis toxin-antitoxin and application of the protein interaction screening system. The protein interaction screening system can realize high-throughput, high-sensitivity and semi-quantitative many-to-many protein interaction analysis by utilizing a flow cytometry sorting technology and three-generation sequencing analysis, effectively solves the problem of false negative caused by non-uniform conversion efficiency of a traditional double-plasmid system, remarkably improves the practicability of high-throughput screening, and has a good application prospect. The large-scale screening bottleneck is broken through, and wide application prospects are achieved.

Provided are a method for freezing fine droplets with which it is possible to more efficiently and appropriately freeze a larger number of fine droplets 15, and a device used therefor. A method and a device for freezing fine droplets whereby fine droplets 15, discharged from a discharge head 11 while including fine matter, are frozen into a vitrified state by cooling rapidly, the fine droplets 15 being frozen by passage through a gas-phase cooling space 20 cooled by a cooling means 30. The cooling space 20 is an interior space in which two ends in the direction of passage of the fine droplets 15 are provided as an inlet and an outlet. A generation device related to the fine droplets 15 including the discharge head 11 is provided by inkjet technology or cell sorter technology. The discharge head 11 is disposed on the inlet 21 side of the cooling space 20 facing the cooling space 20, and a frozen fine droplet receiving part 50 in which the frozen fine droplets are stored is disposed on the outlet 22 side of the cooling space 20.

The invention discloses a kit and a method for preparing a crustacean tissue single-cell suspension, and belongs to the technical field of single-cell suspension preparation. The tissue preserving fluid and the enzymatic hydrolysate in the kit disclosed by the invention cooperate with each other, so that various technical problems of crustacean sample treatment are solved. The single-cell suspension prepared by using the kit and the method disclosed by the invention is high in motility rate and low in caking rate, and can be directly applied to high-end applications such as flow cytometry sorting, primary culture, single-cell sequencing and the like, and the success rate is greatly improved.

The invention relates to the technical field of biomedical engineering, and discloses a method for promoting root rot resistance by analyzing soybean core endophyte flora based on fluorescence labeling, and the method comprises the following steps: S1, respectively separating microbial communities from root systems of healthy soybean plants and susceptible soybean plants to obtain a healthy group microbial suspension and a susceptible group microbial suspension; according to the method for promoting root rot resistance by analyzing soybean core endophyte flora based on fluorescence labeling, phytophthora sojae and associated microorganisms are specifically recognized through an immunofluorescence labeling technology, fluorescence-negative flora in healthy soybean root systems is precisely enriched in combination with flow cytometry sorting, screening leakage of non-culturable microorganisms is avoided from the source, and the yield of root rot resistance is improved. Meanwhile, the invalid workload in the screening process is greatly reduced, the mining period of the core beneficial endophytic flora is shortened, and the pertinence and efficiency of the screening process are improved.

The application discloses an anti-helicobacter pylori monoclonal antibody and a preparation and application thereof, and belongs to the technical field of medical detection. The antibody is prepared by using inactivated helicobacter pylori ATCC 43504 whole bacteria as an immunogen, immunizing a New Zealand white rabbit, and then performing single B cell sorting, limited dilution, gene cloning, plasmid construction and eukaryotic cell expression. The antibody has a specific CDR core sequence, high specificity and high affinity, can accurately recognize helicobacter pylori specific antigens, and effectively avoids cross reactions with other bacteria or tissue impurities. The antibody can be used for detecting helicobacter pylori, and provides a new tool and method for pathological diagnosis of helicobacter pylori infection and related digestive system diseases.

The invention relates to a culture method of TIL from pleural effusion and / or ascites. Specifically, the invention provides the in-vitro culture method of the pleural effusion and / or ascites derived TIL, and the method comprises the following steps: 1) obtaining TIL seed cells from pleural effusion and / or ascites, and 2) sorting the TIL seed cells to obtain intermediate cells which are CD45RA-cells, CD39 + cells, or CD45RA-CD39 + cells.