65 results about "Cell sorting" patented technology

The invention relates to the technical field of biological medicines, and particularly discloses an anti-human CD3 rabbit recombinant monoclonal antibody as well as a preparation method and application thereof. The antibody obtains anti-CD3 immune response by immunizing a rabbit, is obtained by combining single B cell sorting, gene cloning and a mammalian cell expression system, and has high affinity and good specificity. The antibody can effectively recognize CD3 positive T cells in human peripheral blood and can be applied to the fields of flow cytometry, immunohistochemistry, T cell function research and the like. Experimental results show that the antibody has the advantages of high titer, strong fluorescence signal, low background and the like, and is superior to the traditional murine anti-CD3 antibody. Amino acid sequences of light chain and heavy chain variable regions of the antibody are further obtained, and controllability and repeatability of antibody expression are achieved. The antibody has important scientific research value and industrial application prospect, and is suitable for various scenes such as T cell marker detection, immune state evaluation, immunotherapy auxiliary research and the like.

The invention relates to a method for inducing mouse trophoblast stem cells in vitro. According to the method, mouse embryonic stem cells are used as starting cells, Sorcs3 gene knockout mouse embryonic stem cells are obtained through a gene editing technology, and in-vitro induced trophoblast stem cells (iTSCs) are efficiently and rapidly obtained by means of a flow cytometry sorting technology and in combination with specific antibody sorting. According to the invention, the Sorcs3 gene is knocked out from the embryonic stem cell, so that the capability of the embryonic stem cell to differentiate to the trophoblast lineage can be obviously enhanced; an important research platform is provided for deeply researching a lineage intertransformation mechanism of cell masses (ICM) and trophoblast (TE) in the blastocysts, and the method has important significance for researching lineage limitation of early embryonic development of mammals and the like.

The application discloses a flow cytometer sorting control system, a sorting method and a storage medium, and belongs to the technical field of flow cytometer sorting. The flow cytometer sorting control system provided by the application comprises an interface for interacting with user software, an analog-to-digital conversion module, a data processing module, a sorting identification module, a sorting delay module and a sorting actuator. The application further provides a method for sorting cells by using the flow cytometer sorting control system provided by the application, wherein the method comprises a new sorting identification method and a new delay method. By using the control system and the sorting identification method provided by the application, the calculation time can be greatly shortened and the sorting speed can be improved when complex circle gates are used under the same calculation speed of hardware; meanwhile, the time difference between the start of liquid drop charging and the disconnection of the liquid drop can be fixed, the sorting stability and reliability can be ensured, the judgment of cell spacing for each cell is not needed, the judgment of whether each cell is a target cell is not needed, the processing process of cell signals is simplified, and the requirement for electronic performance is reduced.

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.

The invention provides a haematococcus pluvialis freeze-dried algae seed production process which comprises the following steps: algae seed acquisition: automatically separating haematococcus pluvialis seeds from a natural water body water sample by adopting a flow cytometry sorting technology in combination with a fluorescence labeling method; expanding cultivation: adopting an open autotrophy mode, and automatically adjusting the illumination intensity and duration by utilizing an intelligent control system; concentrating, namely concentrating the algae liquid into algae mud through a centrifugal machine; compounding and blending, namely adding a freeze-drying protective agent into the concentrated algae seeds, and uniformly mixing; and filling and storing, namely packaging by adopting nitrogen, and storing in a shady and dry place for a long time. According to the method, flow cytometry sorting is combined with a fluorescence labeling method to separate algae species, the expanding culture environment is intelligently regulated and controlled, a freeze-drying protective agent formula and a vacuum freezing nitrogen packaging process are optimized, the dormant state and long-term stable storage of the algae species are achieved, the storage stability and transportation convenience of the algae species are improved, and meanwhile it is guaranteed that the algae species can be revived with water; and adding a culture medium for propagation and propagation.

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.

The invention provides a method for extracting CD45 positive T cells of mouse placenta decidua and application of the CD45 positive T cells, and belongs to the technical field of tissue cell separation methods.According to the method for extracting the CD45 positive T cells of the mouse placenta decidua, digestive juice and calcium ions with the specific composition are used for the mouse placenta decidua, the concentration of type II collagenase is 0.4-0.6 mg / mL, the concentration of DNaseI is 0.08-0.12 mg / mL, and the concentration of type II collagenase is 0.1-0.5 mg / mL. The concentration of calcium ions is 0.1 to 0.3 mg / mL; then obtaining a cell layer by using a density gradient centrifugation method; carrying out flow cytometry sorting to obtain a CD45 positive cell subset; the extraction efficiency is high, the extracted and separated mouse placenta decidua CD45 positive T cells are small in damage and high in activity, and the extraction method is simple and suitable for industrial application.

The invention discloses a bifidobacterium activity dominant population induction culture method and application thereof, and belongs to the technical field of microbial fermention.The induction culture method comprises the steps that a seed solution is inoculated into a culture medium containing lactose, saccharose or maltose to be subjected to anaerobic fermentation to the end of logarithm; after the fermentation liquor is diluted, SYTOTM9 and PI dye are added, and a flow cytometer is used for sorting. According to the method, lactose, cane sugar or maltose is selected as a carbon source for high-density fermentation, living cells, damaged cells and dead cell subgroups are separated in combination with a flow cytometry sorting technology, and a subgroup differentiation mechanism is analyzed by utilizing non-targeted metabonomics, so that the sugar uptake capacity of the living cell subgroups cultured by cane sugar is improved by 30.55% compared with that of the damaged subgroups; the flowability of the membrane is improved by 95.23% compared with that of maltose culture, the thallus activity is remarkably improved, and a theoretical support and a technical method are provided for industrial production of probiotic high-activity preparations.

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 invention provides an anti-orthopoxvirus monoclonal antibody and application thereof, the monoclonal antibody is obtained by screening through a flow cytometry sorting technology and a single cell PCR technology, has a unique CDR partition, and is mainly used for recognizing L1R (M1R) antigen protein of orthopoxvirus. The affinity of the antibody to VARV (Vector Activated Receptor Virus), VACV (Vector Activated Receptor Virus), CPXV (Cytopoxvirus) and MPXV (Metapoxvirus) of orthopoxvirus is 1.84 nM, 1.46 nM, 2.71 nM and 2.57 nM respectively. In a virus neutralization experiment, the IC50 of the antibody and mature virions in vaccinia virus cells is 37.93 pM. In a monkey pox virus infected mouse model, the antibody can treat a monkey pox virus infected mouse. The monoclonal antibody disclosed by the invention has the characteristics of high expression, high neutralizing activity and good broad spectrum, can be used for industrial production, and can be applied to coping with orthopoxvirus infection outbreak possibly occurring in the future.

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.

A method for enhancing gating performance of a cell sorter to prepare an enriched specimen for optical tomography cell analysis includes introducing a specimen into a FACS to generate 2D event data; generating a first scatterplot of the 2D data; identifying target objects; constructing a boundary within the first scatterplot to produce a first gate; counting target objects within the first gate; comparing the number of target objects within the first gate to a first predetermined value and adjusting the first gate as necessary. A boundary around a set of target objects is constructed in a second scatterplot to produce a subset second gate and target objects within the second gate are counted and the count compared to a second predetermined value. When a boundary around target objects meets specifications the first and second gates are stored in memory and used to enrich patient specimens.

The invention provides an anti-sT2 monoclonal antibody as well as application and a product thereof, and relates to the technical field of biology. According to the invention, a Balb / C mouse and a New Zealand white rabbit are respectively immunized by using sT2 immunogen expressed and purified by a eukaryotic system, and the anti-sT2 monoclonal antibodies anti-sT2-mab1 and anti-sT2-mab2, which are high in purity, high in sensitivity, high in specificity and capable of recognizing different antigen epitopes, are obtained through multi-step screening by using a mouse hybridoma and rabbit B cell sorting method. Compared with the traditional homologous antibody pairing which is easy to cause signal weakening due to epitope steric hindrance, the anti-sT2-mab1 and anti-sT2-mab2 are combined, so that the species cross reaction can be obviously reduced. The antibody developed by the invention can be used for immunological detection of immunoblotting, immunofluorescence and the like.

This invention provides a neutralizing monoclonal antibody B36B9 against orthopoxvirus and its applications. The monoclonal antibody, obtained through flow cytometry sorting and single-cell PCR, possesses a unique CDR region and primarily recognizes the L1R (M1R) antigen protein of orthopoxvirus. The antibody exhibits affinities of 0.21 nM, 0.39 nM, 0.03 nM, and 0.12 nM for VARV, VACV, CPXV, and MPXV, respectively. In virus neutralization experiments, the antibody showed an IC50 value of [missing information - likely related to viral neutralization]. 50 The titer was 41.13 nM. In a mouse model of monkeypox virus infection, the antibody significantly reduced the viral titer in the lungs of mice. The monoclonal antibody provided by this invention has the characteristics of high expression, high affinity, and broad spectrum, and has the potential for application in the preparation of therapeutic and preventive drugs for monkeypox virus or kits for detecting monkeypox virus.

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.

Disclosed is a monoclonal antibody against the Rift Valley fever virus. The antibody is screened and obtained by means of flow cytometric sorting and single-cell PCR technology, and has a unique CDR region. Further disclosed is the use of the antibody in the preparation of a drug for treating Rift Valley fever. The monoclonal antibody against the Rift Valley fever virus has high efficiency and specific activity against the Rift Valley fever virus; and also has the characteristics of high expression, high degree of humanization and good stability, and is suitable for industrial production.

The invention provides an anti-orthopoxvirus neutralizing monoclonal antibody B36B9 and application thereof, the monoclonal antibody is obtained by screening through a flow cytometry sorting technology and a single cell PCR technology, has a unique CDR partition, and is mainly used for recognizing L1R (M1R) antigen protein of orthopoxvirus. The affinity of the antibody to VARV (Vector Activated Receptor Virus), VACV (Vector Activated Receptor Virus), CPXV (Cytopoxvirus) and MPXV (Metapoxvirus) of orthopoxvirus is 0.21 nM, 0.39 nM, 0.03 nM and 0.12 nM respectively. In a virus neutralization experiment, the IC50 of the antibody and mature virions in vaccinia virus cells is 41.13 nM. In a monkey pox virus infected mouse model, the antibody can significantly reduce mouse lung virus titer. The monoclonal antibody provided by the invention has the characteristics of high expression, high affinity and broad spectrum, and has application prospects in preparation of medicines for treating and preventing the monkey pox virus or kits for detecting the monkey pox virus.

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.