111 results about "Cell sorting" patented technology

Provided herein are improved multi-way cell sorter systems and methods. For example, provided are systems and methods for the collection of cells that are sorted in multiple directions. The systems and methods allow for the construction of a multi-way sorter (e.g., a ten-way sorter in the space that currently only allows four-way sorting). In addition the device may actively sense the arrival of drops (with cells of interest) at a sample tube, and trigger an alarm when the drops' arrivals deviate from an expected pattern.

Methods and apparatus are provided for culturing cells under conditions for determining cellular differentiation and for separating cells from culture media based on differentiation. The apparatus comprises a bioreactor, media reservoir, a magnetic cell separator, an inlet port for adding magnetic particles to the bioreactor, and a circulating pump, wherein the bioreactor, media reservoir, magnetic cell separator and inlet port are on a single fluid circuit. The present method and apparatus provides a method for separating cells from culture without removing the cells from culture, so that un-selected cells may be returned to the bioreactor for further culture. The method employs magnetic labeling in culture, where the magnetic label specifically identifies cells to be distinguished, either by separation or retention in the culture. The method and apparatus are further designed to comprise means for electromechanical stimulation of hum embryonic stem cells for preparation of electrically responsive tissue.

Methods and compositions for tracking or monitoring uptake of siRNA by mammalian cells are provided. The methods and compositions may be used to monitoring the silencing activity of the internalized siRNA. The compositions contain an siRNA, an optically or magnetically detectable nanoparticle such as a quantum dot and, optionally, a transfection reagent. Cells are contacted with an siRNA and an optically or magnetically detectable nanoparticle, optionally in the presence of a transfection reagent. Detection of internalized nanoparticles is indicative of siRNA uptake. The invention allows analysis, identification, processing, etc., of cells that have efficiently taken up siRNA. In one embodiment, cells are sorted into at least two populations based on the amount of siRNA taken up.

Prostate-specific membrane antigen (PSMA) targeting compounds are described. Uses of the compounds for imaging, therapy, cell sorting, and tumor mapping are also described.

The invention provides a single-cell-sorting and multigene-locus-detection micro-fluidic chip for expressing EGFR. The micro-fluidic chip comprises a top cover, a micro-fluid channel layer, a substrate and a base plate which are sequentially stacked from top to bottom and sealed with one another, wherein the substrate is provided with micro-pore arrays and the base plate is provided with cavity groups. The invention further provides a system matched with the chip and used for detecting the single cell level of multiple mutation sites of EGFR genes. The micro-fluidic chip and the corresponding detecting system can screen out target cells from a large amount of cells fast, simply and cheaply and perform EGFR multi-gene-site analysis on the single cell in the target cells.

The invention belongs to the field of biotechnology development and application research, and discloses a preparation method for an HLA-A0201 limited antigen specificity CTL (cytotoxic T lymphocyte). The method comprises the following steps: collecting eripheral mononuclear cells one by one, so as to enrich and purify a CD8+T lymphocyte; stimulating the CD8+T lymphocyte with a mature dendritic cell bearing an HLA-A0201 limited target antigen polypeptide, and promoting the growth of the T lymphocyte through the combination of rhIL-2 and rhIL-7; purifying the target CTL according to a Tetramer marking method and a flow cell sorting method; stimulating the growth of the target CTL according to a solid phase coated anti-human-CD3mAb and IL-2; adding an autologous PBMC (peripheral blood mononuclear cell) to enhance the actification of the target CTL; and adding rhIL-15 for enlarging cultivation, collecting and identifying. The CTL prepared according to the method has the advantage that the purity, the proliferation capability, the cytotoxicity and the CTL-CM proportion are high, so as to be used for immunological therapy of tumors and the like.

The invention belongs to the technical field of cell culture in vitro, and particularly relates to a preparation method of high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The method comprises the following steps: collecting and separating peripheral blood mononuclear cell of a patient, eliminating CD4+CD25+Treg cell by means of Mini MACS (magnetic active cell sorting) method, and sorting to obtain CD3+, CD4+ and CD8+T cells; and putting the obtained cells into culture solution containing phytohemagglutinin (PHA), so that the PHA concentration in the suspension liquid is 100ng / ml, hatching for 24h under the culture condition of 5% CO2 at 37 DEG C, transferring the hatched suspension liquid into a cell culture bottle coated by CD3 monoclonal antibody (1mug / ml), adding IFN (interferon)-gamma (1000U / ml), adding IL (interleukin)-2(500U / ml) and IL (interleukin)-21(1000U / ml) after 48h, compensating sodium selenite-containing (0.005mg / L)cell culture after four days, and continuously culturing for 7-14 days to obtain the high-purity, high-multiplication capacity and high-cytotoxin activity CIK (cytokine induced kill) cell. The quantity, the activity and the purity of the CIK cell which is prepared by the method and amplified in vitro are improved, so that the antineoplastic function of the CIK is enhanced.

The invention provides a high-integration equidistance equipartition nucleic acid amplification micro-fluidic chip. The high-integration equidistance equipartition nucleic acid amplification micro-fluidic chip sequentially consists of a cover glass layer, a reaction layer, a decorative layer and a sealing layer from bottom to top in a thermal bonding manner, wherein the reaction layer is provided with n reaction modules, and each reaction module is provided with a circulation passage, a reaction micro-cavity and a sample feeding opening by utilizing a multilayer soft photoetching technology. By adopting the chip, the circulation distance of the sample before being introduced into a small chamber is identical, the sample feeding quantity is identical, and an error caused by the non-uniformity of the reaction volume can be avoided. The experiment cost is reduced. The sample feeding speed is increased. Quantitative detection in real time can be realized. The chip is reasonable in design, simple in structure, simple to operate and low in production cost and can be applied in the real-time fluorescent quantitative nucleic acid amplification, digital nucleic acid amplification, single-cell separation of nucleic acid amplification, SNP detection, detection of single-base mutation, detection of copy number imbalance, research of single-cell gene expression profile, early detection of cancer marker, stem cell differentiation identification, cell separation identification and the like.

An object is to provide a TCR closing system that enables not only bias-free analysis of TCR repertoires, but also collection of antigen-specific TCR α / β cDNA pairs and evaluation of functions thereof. There is provided a method for producing a gene of T cell receptor (TCR) specific to an antigen A, which comprises 1) the step of stimulating a group of T cells including a T cell specific to an antigen A or one T cell specific to an antigen A under a condition effective for amplification of a TCR gene; 2) the step of identifying a T cell specific to an antigen A among the group of T cells including a T cell specific to the antigen A, and sorting one T cell specific to the antigen A into a vessel; and 3) the step of subjecting the one activated T cell specific to the antigen A in the vessel to PCR to amplify a gene of TCR specific to the antigen A. According to the present invention, a target TCR gene can be cloned within a shorter time compared with that repaired by the conventional methods, for example, about ten days. Further, according to the present invention, genes of TCR α chain and β chain can be highly efficiently cloned. Under the conditions of the examples, a pair of a TCR α chain and TCR β chain could be obtained from stimulated T cells sorted as single cells at a ratio of 100%.

The invention provides a plurality of embryonic stem cell-like progenitor cells, which are isolated from a human tissue by a systemic screening of human mesenchymal stromal stem / progenitor cells and a cell sorting by a cell antigen selected from the group consisting of CD34, CD117, CD133, CD201, GloboH and combination thereof, and cultured in a medium supplemented with at least one or more steroids and one or more growth factors. The cells of the invention express CD34 and exhibit sphere-like clonogenicity in early passages and express multipotent embryonic stem cells (ESCs) like characteristics.

The invention discloses polypeptide for specifically targeting a lung cancer cell and an application thereof. The polypeptide for specifically targeting the lung cancer cell has an amino acid sequence shown as SEQ ID NO.1, and is prepared by screening on the surface of the human lung cancer cell by an ex-vivo bacterial random polypeptide library exhibiting method and a flow cell sorting method. The polypeptide for specifically targeting the lung cancer cell can be applied to preparation of lung cancer diagnosing tracer, lung cancer cell detecting reagent and the like. The polypeptide for specifically targeting the lung cancer cell can be combined with a lung cancer A549 cell specifically, is not combined with a normal lung cell and other tumor cells; and the preparation method is simple, feasible and applicable to large-scale industrial production. The polypeptide for specifically targeting the lung cancer cell provides important theoretical and practical basis for early diagnosis, targeting therapy of the lung cancer and the like, and has a wide application prospect.

The cell-delivery unit of a high-throughput electrophysiological testing system is implemented as a reusable movable unit suitable for repetitive delivery of cells to a disposable, multi-aperture patch-clamp tray. An electric field emanating from the patch aperture is used to align the dispenser with the aperture. A set of electrodes in the nozzle of the dispenser is used to detect the electric field and effect the alignment. According to another aspect of the invention, dielectrophoretic fields produced by sets of electrodes in the nozzle form a retaining cage that is used first to suspend a test cell directly above the patch aperture and then to urge the cell toward it. A movable cell sorter may also be coupled to the cell-delivery unit.

The invention discloses a method for rapidly acquiring nano-antibodies. The method comprises the following steps: (1) acquiring a negative control; (2) separating peripheral blood mononuclear cells (PBMC) of an alpaca; (3) carrying out flow cytometry sorting; (4) extracting RNA of positive cells, and carrying out inverse transcription to form cDNA; (5) amplifying VHH segments; (6) linking the VHH segments into an expression vector; (7) carrying out PCR identification on a recombinant prokaryotic expression vector; (8) carrying out expression of pet28a-CD19-VHH; and (9) carrying out ELISA activity identification on the expression vector.

The invention discloses an efficient genetic-modification-free iPSC induced and industrialized monoclonal picking platform and application. The platform can efficiently perform reprogramming and only needs to use a minimum number of reprogramming factors (OCT4, SOX2, E6 and E7). In a monoclonal separation stage, SSEA4 / TRA-1-60 is taken as a selection marker, a large number of single cell clones are obtained through a flow cytometry sorting technology. The platform has the advantages of being high in reprogramming efficiency, high in safety, easy to operate, capable of achieving large-scale production and the like.

The invention discloses a micro-fluidic chip detecting system and method for sorting and focusing of integrated cells. The micro-fluidic chip detecting system comprises a viscoelastic fluid sampling device, a sample sampling device, a waste liquid collecting device, a high-speed image acquiring device, a micro-fluidic chip, a sample collecting device and a microcomputer, wherein the micro-fluidicchip is manufactured by encapsulating of a flow channel layer and a basal layer; and a spiral flow channel, a branch flow channel, a lower division flow channel, an upper division flow channel, a viscoelastic fluid sampling flow channel, a spiral mixing flow channel and a focusing main flow channel are arranged in the flow channel layer. Through the adoption of the micro-fluidic chip detecting system disclosed by the invention, cells are sorted according to the dimension characteristics, the viscoelastic bearing fluid is mixed and blended, and the sorted rare cells are focused in single equilibrium positions at the centers of the sections of the flow channels, so that the accuracy of counting and detection is improved.

The invention relates to a preparation method of CIK (Cytokine Induced Killer) cells with high proliferation capacity, high cytotoxic activity and high survival rate. The preparation method comprises the steps of: (1) sorting and removing CD4<+>CD25<+>Treg cells of peripheral blood mononuclear cell to obtain CIK pre-cells; (2) cultivating the CIK pre-cells in a cell culture fluid containing 100ng / ml of PHA, 100ng / ml of IL-6 and 10ng / ml of PGE2 for 24h; (3) transferring the CIK pre-cells to a cell culture bottle coated with 1microgramme / ml of CD3 monoclonal antibody, and adding 1000U / ml of IFN-gamma for cultivating for 48h; (4) adding 1000U / ml of IL-2 and 100ng / ml of IL-1alpha for cultivating for 4 days; and (5) adding 1microgramme / ml of insulin to continuously cultivate for 7-14 days. The invention further provides associated CIK cells, a method for inhibiting the peripheral blood mononuclear cell to be differentiated to the CD4<+>CD25<+>Treg cells and a method for promoting the proliferation of the CIK cells. The preparation method of the CIK cells provided by the invention is skillful in design, and the tumor killing cells-CIK cells prepared have stronger proliferation capacity, higher cytotoxic activity and better tumor killing efficiency, so that the clinical efficacy is improved, and the preparation method is appropriate for wide application in a large scale.

The invention provides a novel high throughput screening model for hepatitis C virus (HCV) resistant drugs. The high throughput screening model can be applied to research and drug screening of human HCV infection mechanisms, and is a reporting system carrying out efficient response aiming at HCV infection. After the HCV invades cells, protease NS3 / 4A expressed by the HCV is about to cut and release a tetracycline-regulated transactivator (rtTA) anchored on a mitochondrial outer membrane by an MAVS (mitochondrial antiviral signaling) protein, expression of a reporter gene is activated in the presence of doxycycline or tetracycline, and obvious fluorescence signals are observed in a certain time; or cell death is caused in the presence of GCV (gancilovir). The system is simple to operate, has short experimental period, high efficiency and strong stability, can be widely applied to real-time quantitative observation of HCV infection, flow cell sorting, high throughput compound screening, high throughput host cell target gene screening and molecular mechanism research of HCV infection.

The invention relates to a dielectrophoresis micro-fluidic chip for selecting and focusing cells as well as an alignment-free microprocessing method thereof, which belongs to the technical field of micro-fluidic chip. The dielectrophoresis micro-fluidic chip comprises a substrate layer, an integrated runner-electrode functional layer and a top layer, whereinthe integrated runner-electrode functional layer comprises a main body electrode, an interdigital electrode extending into a main runner from a main electrode and perpendicularly crossing the main runner, a main runner inlet, a main runneroutlet, parallel branch runners formed by dividing the main runner by virtue of non-uniform side wall structures of the interdigital electrode and an insulation channel. High-performance dielectrophoresis forces in the horizontal direction and vertical direction are generated by virtue of the interdigital electrode formed by multiple layers of electrodes and having the non-uniform side walls so asto improve the separation purity, the cell focusing can be realizedat the same time, and the flux can be improved by virtue of the parallel runners formed by the non-uniform side walls; and an SU-8 mold with a multilayer structure is filled in one step by using a homogenous conductive polymer, and the integrated runner-electrode structure can be formed in one step by reversing the mold.

The invention discloses an optical flow control cell sorting chip and method. The chip comprises a substrate, a microfluidic channel and an optical waveguide. The microfluidic channel is disposed in the substrate, and the optical waveguide is movably disposed on the substrate. The microfluidic channel comprises at least a first channel and a second channel which are mutually communicated, the first channel and the second channel are both used for accommodating microfluidics, and the first channel is further used for accommodating at least two kinds of cells. The optical waveguide comprises anoptical signal input end and a capture end. The optical signal input end is used for accessing an optical signal, the capture end is used for adsorbing one of at least two kinds of cells, and the capture end is used for releasing the adsorbed cells to the second channel. The chip is simple in structure and manufacturing method, low in cost, capable of sorting micro-nano cells accurately efficiently and flexibly by using the cone waveguide and capable of separating unmarked biological cells.

The invention discloses a lithium battery cell sorting device. The lithium battery cell sorting device comprises a discharge mechanism, an electrical performance testing mechanism and a sorting mechanism. The lithium battery cell sorting device is characterized in that the discharge mechanism comprises a discharge hopper and a conveyor belt; a vertical discharge tube is formed on the lower portionof the discharge hopper; a screw rod is vertically arranged in the discharge tube; a rotary disk is arranged at the middle end of the conveyor belt; the electrical performance testing mechanism comprises a base, a first feeler pin and a second feeler pin; a battery locating groove is formed in the base; the sorting mechanism comprises a sorting tube; the sorting tube vertically communicates witha plurality of sorting channels; a material receiving plate is slidably arranged on the upper portion of each sorting channel; the lower portion of each sorting channel is provided with a material collection assembly; each material collection assembly comprises a material collection platform, a material box rack, a material box, a material limiting plate and a material baffle; and a pushing cylinder is arranged on each material collection platform, and the output end of the pushing cylinder is provided with a pushing plate opposite to the corresponding material limiting plate. The lithium battery cell sorting device provided by the invention can complete sorting operation of lithium battery cells and effectively improve the production efficiency.

The invention discloses a double-layer microfluidic chip for tumor cell sorting, comprising a first chip and a second chip positioned below the first chip; the surface of the first chip has a sample passage, the surface of the first chip facing the sample passage has a buffer passage, and the sample passage is cross-communicated with the buffer passage. Compared with CellSearch detection method, the technique of using the double-layer microfluidic chip to analyze tumor cells can be finished continuously in one step with no need for multiple steps and is simple to perform, detection speed is high, the purity of enriched CTCs is higher, no antibody is used, and CTCs obtained by the technique have high cell activity and may be better used in subsequent detection and tumor specificity research.

The invention discloses a humanized anti-H7N9 avian influenza virus high-affinity antibody 10K filtered and obtained based on a single cell separation technology, the amino acid sequences of light chain and heavy chain variable regions of the antibody are shown as SEQ ID No. 2 and SEQ ID No. 5 respectively. The high-affinity specificity of the antibody is combined with H7N9 avian influenza virus 7 type hemagglutinin protein, and can mediate the kill and wound (ADCC) of effector cells using NK cells as main parts for H7N9 influenza virus infected cells. The antibody 10K can be used for therapeutic development of highly pathogenic avian influenza infection, and also can be used for development of H7N9 influenza virus antigen dectection reagents.

The invention relates to a B cell screening method and application of the B cell screening method in preparing a monoclonal antibody. The B cell screening method comprises the steps of S11, adding a signal peptide sequence of secretory expression at the N end of a Protein A protein gene, and adding a membrane-spanning gene sequence at the C end to obtain a fused exogenous gene; S12, using the fused exogenous gene to conduct transfection on a B cell of which immunity is completed, and adopting the B cell as a cell to be screened; S13, adding an antigen for labeling fluorescent into the cell to be screened, using a flow cytometry to sort and collect cell carrying fluorescent to obtain a positive B cell. According to the B cell screening method and application of the B cell screening method in preparing the monoclonal antibody, antibody molecules are gathered on the surface of the cell, through a fast and precise sorting function of the flow cytometry, a cell to be fused is screened in advance, thus the success rate and accuracy rate of a traditional mode are sharply increased, and the problem of detection of a monoclonal antibody cell strain in a later period is solved at the same time.

Provided herein are improved multi-way cell sorter systems and methods. For example, provided are systems and methods for the collection of cells that are sorted in multiple directions. The systems and methods allow for the construction of a multi-way sorter (e.g., a ten-way sorter in the space that currently only allows four-way sorting). In addition the device may actively sense the arrival of drops (with cells of interest) at a sample tube, and trigger an alarm when the drops' arrivals deviate from an expected pattern.

The invention belongs to the technical field of biochemical separation and analysis, and particularly relates to a micro-fluidic chip, a system and a method for sorting and enriching cells in cerebrospinal fluid. The chip can avoid pretreatment operations such as sample pretreatment and cell labeling and realize fast and efficient sorting and enriching functions for a small quantity of cells in the cerebrospinal fluid and further can realize in-situ counting and fast distinguishing of the small quantity of cells in the cerebrospinal fluid in combination with an in-situ microtechnique. The system integrates fast and efficient separation and enrichment of the small quantity of cells in the cerebrospinal fluid and in-situ microscopic observation, can realize sorting, enrichment, counting, observation and detection for the small quantity of cells in the cerebrospinal fluid, is high in functionality and high in efficiency, and can be used for clinical fast and efficient detection for cerebrospinal fluid cells.

The invention provides a method for rapidly acquiring a nano antibody by using a high-flux sequencing technique. With the combination of a flow cytometry separation technique and a high-flux sequencing technique, the method comprises the following steps: acquiring alpaca peripheral blood mononuclear cells (PBMC) which are immunized or unimmunized by using a specific protein W; acquiring PBMCs which are specified for the specific protein W by using a flow cytometry separation method; extracting total RNA (Ribonucleic Acid) of unimmunized, immunized and separated PBMCs, and performing inverse transcription into cDNA (Complementary Deoxyribonucleic Acid); by taking the cDNA as a template, performing two-step method PCR (Polymerase Chain Reaction) amplification so as to obtain DNA fragments of a specific protein W resistant nano antibody; establishing a DNA library of the DNA fragments of the nano antibody, and performing high-flux sequencing analysis; and analyzing the abundance of the DNA fragments of the nano antibody; establishing nucleotide sequences of the DNA fragments of the nano antibody into an expression vector, and expressing the DNA fragments of the nano antibody in appropriate hosts.

The invention discloses a cancer cell or other pathologic cell detection diagnostic device which is composed of a laser source system, a beam shaping system, a laser confocal system, a beam splitting system, a microscope system, a cell flow system or a cell acquisition system, a fluorescence focusing system and a cell analysis system. The laser source system emits laser with different wavelengths and the laser is shaped by the beam shaping system; the focuses are gathered on the same plane by the laser confocal system after the laser is shaped; then the laser enters the microscope system through the beam splitting system and is irradiated on stained cells in the cell flow system or the cell acquisition system; the fluorescence generated by the stained cells after irradiation of the laser is amplified by the microscope system and then enters the beam splitting system; and the amplified fluorescence is reflected to the fluorescence focusing system by the beam splitting system and thereafter enters the cell analysis system for cell separation to obtain separation results. The cancer cell or other pathologic cell detection diagnostic device disclosed by the invention is high in detection speed and high in precision, and can be applied to early diagnosis, early treatment and prognosis of cancer cells and other pathologic cells.

The invention discloses a preparation method of an HLA-A0201 (Human Leukocyte Antigen-A0201) restrictive anti-MAGE (Micro Array and Gene Expression) antigenic specificity CTL (Cytotoxic T Lymphocyte). The method comprises the following steps of: carrying out hemapheresis to collect peripheral blood mononuclear cells; enriching and purifying CD8+T lymphocytes; utilizing a mature dendritic cell loaded with an HLA-A0201 restrictive anti-MAGE antigen polypeptide to stimulate the CD8+T lymphocytes; utilizing rhIL-2 and rhIL-7 to accelerate the T cells to grow; utilizing a Tetramer marking method and a flow type cell separation technology to purify the target CTL; utilizing an anti-human CD3 monoclonal antibody covered by a solid phase and an IL-2 to stimulate the target CTL to grow; adding an autologous PBMC (Peripheral Blood Mononuclear Cell) radiated by gamma rays to enhance the activation of the target CTL; adding rhIL-15 to cultivate and reproduce; and collecting and identifying. The prepared target CTL has high purity, high reproduction capability, high killing activity and high ratio CTL-CM, and can be used for immune treatment of tumors and the like.

The invention discloses a detection method of the titer of a lentivirus carrier, which particularly comprises the following steps: detecting the titer of a control virus by a flow cytometric sorting method, allowing the virus to be detected and the control virus to infect target cells with the same amount under the same condition, extracting total DNA of the target cells infected by the virus to be detected and the control virus respectively, obtaining a ratio R of the relative contents of the genomes of the virus to be detected and the control virus in the genomes of the target cells by relative quantification PCR detection; multiplying the obtained titer of the control virus with the ratio R of the relative contents of the virus to be detected and the control virus to obtain the titer of the virus to be detected which is the titer of the recombinant lentivirus. The invention is applicable to the titer test of a recombinant lentivirus without marker genes, and can accurately determine the titer of the recombinant lentivirus against target cells which has less amounts and can not be largely cultured and propagated in vitro.