76 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 provides a multi-channel viscoelastic micro-fluidic chip, a manufacturing method and a method for separating MCF-7 cells. The multi-channel viscoelastic micro-fluidic chip comprises a first multi-stage shunting module, a pipette, a second multi-stage shunting module, a single-stage viscoelastic micro-fluidic sub-module and a third multi-stage shunting module, the first multi-stage shunting module is connected with one inlet of the single-stage viscoelastic micro-fluidic sub-module through a pipette; the second multi-stage shunting module is connected with the other inlet of the single-stage viscoelastic micro-fluidic sub-module; and the outlet of the single-stage viscoelastic micro-fluidic sub-module is connected with the third multi-stage shunting module. According to the method, the PEO solution is used as sheath fluid, and the action of elastic force is introduced to cells, so that CTC can be directly separated from undiluted human blood in an ultrahigh-flux manner, and the limitation that erythrocytes need to be split and a blood sample needs to be diluted by dozens of times before cell separation in traditional inertial microfluidics is broken through; and the complexity of operation and the risk of sample pollution are reduced.

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 invention provides a method for purifying ovarian progenitor cells and application of the ovarian progenitor cells to in-vitro generation of steroid secretion cells, and belongs to the technical field of biology. The method for purifying the ovarian progenitor cells comprises the following steps: digesting, filtering and washing an ovary, purifying through a magnetic bead or flow cytometry sorting procedure, and screening to obtain the ovarian progenitor cells, a marker used in the purification process is LY6A, LGR5 or PDGFRA (Platelet Derived Growth Factor Receptor A). Experimental results show that the PDGFRA < + > progenitor cells and the LY6A < + > progenitor cells can form androgen synthetic cells for expressing CYP11A1 and CYP17A1; and the LGR5 < + > progenitor cells can only form progestational hormone synthetic cells which express CYP11A1 but do not express CYP17A1.

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 present invention discloses a method for high-purity sorting and capturing of rare cells. The method for high-purity sorting and capturing of rare cells in the present invention first spreads the cells, then locates and determines the position of the target cells, then uses a probe to expel the buffer outward to disperse the non-target cells around the target cells, and then uses hydrodynamic force to capture the target cells without damage. Finally, the target cells can be released into a receiving container to achieve high-purity sorting and capturing of rare cells. The device used in the method of the present invention has a simple structure and is easy to operate. The method has the advantages of not requiring repeated large-scale dilution, fast spreading speed of cell samples, high sorting purity of target cells, low omission rate, in-situ capture, and good repeatability, and is applicable to fields such as single-cell screening and analysis, single-cell culture, single-cell drug screening, single-molecule analysis, gene screening, sample purification, antibody screening, and microbial research.

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.

The disclosed invention presents the AutoCell Platform (ACP), an advanced, fully automated, and functionally closed system designed to revolutionize the manufacturing of gene-modified cell therapies. This innovative platform integrates automation, closed-loop processing, and novel technologies to address the inefficiencies and high costs of conventional methods. Key features include automated spinoculation for enhanced genetic modification efficiency, microbubble-assisted cell selection for precise cell isolation, and modular design for scalability across clinical and research settings. By reducing production timelines from 30-40 days to under 3 days and lowering costs by an order of magnitude, the ACP enables point-of-care manufacturing, decentralization, and broader accessibility. Advanced quality control measures and a standardized design ensure regulatory compliance and consistent therapeutic outcomes. The ACP supports diverse applications, from CAR-T therapies to regenerative medicine, representing a transformative leap in precision medicine and the global accessibility of life-saving cell-based therapies.

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.

Glycan-binding proteins, and compositions thereof, are generally described, including methods of making and using such proteins. The proteins may include scaffolds based on easily evolvable DNA-binding proteins, with binding sites able to specifically bind to mono- or disaccharides, such as monosaccharide-binding determinants, disaccharide-binding determinants, more complex carbohydrates, etc. In certain aspects, a protein may be generated starting from a small DNA-binding protein, such as Sso7d. Such glycan-binding proteins may have numerous applications, including in enzyme-linked immunosorbent assays (ELISAs), glycan characterization, cell selection, flow cytometry, histology, imaging, arrays, affinity purification, enzyme-linked visualization, binding to a target for pharmaceutical purposes, etc.

The invention provides a cell elution method, equipment and a medium, and the cell elution method comprises the following steps: after a magnetic field is confirmed to be generated in a magnetic separation column, controlling a sample liquid to flow through the magnetic separation column so as to adsorb magnetic labeled cells in the sample liquid into the magnetic separation column through the magnetic field; performing gas-liquid cleaning operation on the magnetic separation column for a first preset number of times; the gas-liquid cleaning operation comprises the following steps: controlling a first preset dosage of gas to clean the magnetic separation column at a first preset flow rate, and then controlling a second preset dosage of buffer solution to clean the magnetic separation column at a second preset flow rate, so as to remove impurities except the magnetically labeled cells in the magnetic separation column through gas-liquid alternate cleaning. According to the magnetic separation column, the magnetic labeled cells can be successfully separated, the purity and the separation precision of the magnetic labeled cells adsorbed by the magnetic separation column are also improved, and the cell separation efficiency is further improved.

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.