217 results about "Sorted Cells" patented technology

The invention relates to the field of cell analysis, and provides a multi-mode digital micro-fluidic system and method for cell sorting analysis, the system comprises an active digital micro-fluidic platform, the active digital micro-fluidic platform comprises a micro-fluidic chip, an upper computer control system and an FPGA hardware circuit, an upper computer generates a liquid drop path planning instruction, the FPGA hardware circuit converts the liquid drop path planning instruction into an electrode driving signal, and the electrode driving signal is transmitted to the micro-fluidic chip; the nanoliter-grade liquid drops are driven to change between the electrodes; the optical tweezers platform comprises a laser device, an optical microscope and a CMOS camera, laser beams are generated by the laser device to form a three-dimensional optical potential well, the laser beams are transmitted into the micro-fluidic chip through the optical microscope to realize non-contact control on cells, and the CMOS camera records images and transmits the images back to the upper computer; the objective table clamp is used for realizing cooperative control of the optical tweezers and the active digital microfluidics; and the image recognition system is used for recognizing morphological characteristics of cells in the liquid drops based on a convolutional neural network and screening target cells. According to the invention, integration of high-throughput nondestructive cell sorting and in-situ multi-modal analysis is realized.

The invention discloses an NKT cell culture optimization method, and belongs to the technical field of cell culture.The method comprises the steps that firstly, PBMC is sorted through a spiral micro-fluidic chip integrated with CD3 / CD56 double-labeled magnetic beads, and CD3 + CD56 + cells with the purity larger than or equal to 95% are obtained through the synergistic effect of dielectrophoretic force and a magnetic field; then, in a GIBCO culture solution added with the IL-2 / IL-15 / alpha-GalCer sustained release microspheres, the pH, dissolved oxygen and lactic acid concentration are accurately regulated and controlled through a real-time monitoring system, and temperature-oxygen partial pressure gradient culture is adopted; and finally, three times of centrifugation are combined with NKT4 solution treatment, and the cells are collected in cooperation with a gradient cooling procedure, so that the cell activity maintenance rate is greater than or equal to 95%. Compared with the traditional technology, the method has the advantages that the NKT cell sorting purity and activity are remarkably improved, the culture environment is regulated and optimized through dynamic metabolism, and accurate control over the culture process is achieved.

The present disclosure relates to automated systems and methods adapted to quickly and accurately train a neural network to detect and / or classify cells and / or nuclei. The present disclosure also relates to automated systems and methods for using a trained cell detection and classification engine, such as one including a neural network, to classify cells within an unlabeled image.

The invention provides a flow sorter liquid drop delay adjusting method and system based on real-time cell flow velocity, relates to the field of flow sorting, and ensures that the liquid drop delay time can not be delayed even if factors such as temperature, nozzle state, air pressure and pipelines are changed in the sorting process by monitoring the cell flow velocity in real time and dynamically adjusting the liquid drop delay time. And the charge loading time of the liquid drop can be accurately controlled. Meanwhile, the liquid drop delay time is dynamically adjusted in real time, so that the random distribution of the cell flow velocity during high-speed sample loading can be effectively dealt with, and the cells at different positions can be accurately sorted. The automatic calibration and self-stabilization functions are achieved, a large number of experiment articles and a complex optimization process are not needed, accurate control over liquid drop delay can be completed only through real-time monitoring and adjustment on the basis of liquid drop delay initial value setting, the separation efficiency is improved, the experiment cost and the operation difficulty are reduced, and the separation efficiency is improved. And the cell sorting requirements in various biomedical fields are met.

The invention provides a marker of spinal cord functional neural precursor cells, a detection method and a sorting method, and belongs to the technical field of biological medicine. The invention provides application of a marker detection reagent in preparation of a kit for detecting spinal cord functional neural precursor cells. The invention further provides a sorting-based method for separating the SFNPCs cells, and the cells are sorted through the Marker of the specific SFNPCs, so that the cell purity of the SFNPCs is remarkably improved, the difference between batches is reduced, and a high-purity and homogeneous SFNPCs cell population is obtained. The survival rate of the SFNPCs cells separated and subcultured to the P9 generation by the method disclosed by the invention is more than 85%, and the SFNPCs cells have a good treatment effect on a spinal cord injury model.

The embodiment of the invention discloses a magnetic separation chip for cell separation and a preparation method thereof.The magnetic separation chip structurally comprises a sample layer, a functional layer, a clamp set and a magnetic field source, the sample layer is composed of a flow channel layer and a sealing film, and the flow channel layer is provided with a sheath fluid inlet, a sample inlet and a sample outlet; the sealing film is attached to the open surface of the runner layer to seal the runner; the functional layer is located below the sample layer and comprises a patterned substrate and an amorphous alloy soft magnetic strip material, the patterned substrate is provided with a counter bore groove, the amorphous alloy soft magnetic strip material is embedded into the counter bore groove, and the upper surface of the amorphous alloy soft magnetic strip material is flush with the upper surface of the substrate, right faces the sealing film and is arranged in the extending direction of the flow channel; the clamp group comprises an upper clamping plate and a lower clamping plate, the upper clamping plate is arranged above the sample layer, the lower clamping plate is arranged below the functional layer, the upper clamping plate is provided with runner holes corresponding to the inlets and the outlets, and the two are connected through bolts to clamp and fix the sample layer and the functional layer; the magnetic field source is arranged at the end where the sample outlet is located and on one side opposite to the functional layer, so that a gradient magnetic field which is gradually enhanced from the inlet to the outlet is formed in the flow channel.

The invention relates to a Raman activated cell sorting system coupled with micro-fluidic and Raman optical tweezers and a method thereof, and relates to the technical field of cell sorting, the system comprises a micro-fluidic chip with low Raman and fluorescence background noise and a pneumatic micro-valve; in the micro-fluidic chip, a sample channel and a micro-cavity are connected with a main channel, and a flow guide channel is connected with the micro-cavity; the flow guide channel and the collecting channel are connected through a pneumatic micro valve. After the pneumatic micro valve is closed, the fluid in the micro cavity is static, and a narrow sample flow is formed in the main channel. And when the Raman collection time is less than the time difference of two adjacent cells flowing through the capturing point of the optical tweezers in the narrow sample flow, high-flux Raman detection can be performed in the narrow sample flow. On the contrary, the target cells can be moved out of the narrow sample flow, and high-precision Raman detection in a long Raman collection scene can be realized. After sorting is completed, fluid flowing of the main channel is stopped, the pneumatic micro valve is opened, the cells stored in the micro cavity are released and collected in a unified mode, the cell suspension volume is more stable and uniform, and follow-up experiment operation is facilitated.

The invention provides a cell magnetic sorting column and a preparation method thereof, and belongs to the technical field of cell magnetic sorting, the method comprises the following steps: S1, pretreating balls; s2, coating the surface of the ball with a thermoplastic high polymer material layer by adopting an electro-deposition process to form magnetic matrix particles; s3, sequentially putting a lower-layer filter screen, magnetic matrix particles and an upper-layer filter screen into the bottom shell of the separation column; s4, the upper sealing cover and the bottom shell are connected through ultrasonic welding; s5, centrifuging the sorting column subjected to ultrasonic welding; and S6, curing the centrifuged separation column. The cell magnetic separation column and the preparation method thereof are particularly suitable for magnetic separation of nanoscale magnetic beads, the problems that a traditional separation column is high in magnetic bead wastage rate, uneven in magnetic field, prone to blockage, prone to cell damage and the like can be solved, a high-purity and high-activity cell source is provided for cell therapy such as CAR-T, and the cell magnetic separation column is suitable for magnetic separation of nanoscale magnetic beads. And the process is simple and suitable for large-scale standardized production.

The invention discloses a cell sorting micro-fluidic chip and application, the micro-fluidic chip is obliquely arranged on an inclined plane, the micro-fluidic chip comprises a substrate, the substrate is provided with a slender groove with a triangular prism profile, and a micro-fluidic channel is embedded in the slender groove; the upper end and the lower end of the inclined surface respectively face the center and the edge of the turntable; one side of the microfluidic channel back to the substrate is parallel to the upper surface of the substrate, one end of the microfluidic channel is communicated with the fluid buffer cavity, the other end is communicated with the sorting channel, and the other end of the sorting channel is communicated with the plurality of sorting sub-channels; a soft magnet is further arranged on the microfluidic channel, and a permanent magnet is arranged on one side, back on to the microfluidic channel, of the soft magnet. According to the invention, two-parameter sorting of preferential separation of magnetic microspheres according to density and accurate positioning according to size can be realized, high-flux and high-precision two-parameter sorting is realized, the resolution is greatly improved, the energy consumption is low, and the device is suitable for portable equipment.

The invention discloses a method for screening and identifying ribosome entry sites in a short chain, which comprises the following steps of: (1) screening a short chain IRES sequence with the length of less than 100nt from an IRES database, constructing a screening library and synthesizing; (2) constructing a screening vector containing double reporter genes, cloning the short-chain IRES into a linearized screening vector, and generating circular RNA which only depends on the short-chain IRES sequence to recruit ribosome expression of the reporter genes in cells by the screening vector cloned with the short-chain IRES sequence; and (3) carrying out cell transfection on the screening vector cloned with the short-chain IRES sequence, screening out cells expressing reporter genes through cell sorting, and sequencing through a ribosome-newborn peptide chain compound to obtain the short-chain IRES sequence capable of recruiting ribosome to start circular RNA translation in the cells. According to the invention, a short-chain IRES sequence library is constructed, and short IRES capable of starting circular RNA translation is screened, so that transfection limitation caused by overlarge molecular weight of the current IRES sequence is overcome.

The application provides a preparation method of a multi-layer sandwich type epoxy group modified magnetic microsphere and application thereof. The multi-layer sandwich type epoxy magnetic microsphere prepared by the preparation method provided in the application can realize efficient and accurate cell sorting through coupling of an antibody, is suitable for cell sorting related scenes such as stem cell separation, tumor cell detection, immune cell treatment (such as CAR-T cell preparation) and pathogenic microorganism enrichment, and provides key material support for cell sorting technology in life science research and clinical diagnosis and treatment.

In alternative embodiments, provided are self-activating Förster resonance energy transfer (saFRET) biosensors, and methods for making and using them. In alternative embodiments, provided are self-activating FRET (saFRET) biosensors, and methods that couple FRET and sequencing (FRET-Seq) to integrate random mutagenesis, fluorescence-activated cell sorting (FACS), and next-generation sequencing (NGS) to screen and identify sensitive biosensors from large-scale libraries directly in mammalian cells, utilizing the design of saFRET biosensors as provided herein.

The application relates to the technical field of cell sorting, and specifically provides a feature fusion method and system for high-throughput cell sorting. The method comprises the following steps: acquiring real-time image feature distribution deviation according to real-time image feature distribution and reference image feature distribution, and acquiring real-time spectral feature distribution deviation according to real-time spectral feature distribution and reference spectral feature distribution; calibrating all real-time image features according to the real-time image feature distribution deviation, and calibrating all real-time spectral features according to the real-time spectral feature distribution deviation; for each cell to be sorted, fusing real-time image features and real-time spectral features to obtain a fusion feature vector, and then classifying the cell to be sorted according to the fusion feature vector; the method can effectively improve the long-term stable operation capability, performance repeatability, reliability in standardized application scenarios and reduce the complexity and time consumption of system calibration and troubleshooting of a high-throughput cell sorting system.

The invention relates to the technical field of cell sorting, and particularly provides a feature fusion method and system for high-throughput cell sorting, and the method comprises the steps: obtaining a real-time image feature distribution deviation according to real-time image feature distribution and reference image feature distribution, acquiring a real-time spectral feature distribution deviation according to the real-time spectral feature distribution and the reference spectral feature distribution; calibrating all real-time image features according to the real-time image feature distribution deviation, and calibrating all real-time spectral features according to the real-time spectral feature distribution deviation; for each to-be-sorted cell, fusing the real-time image feature and the real-time spectral feature to obtain a fused feature vector, and then classifying the to-be-sorted cells according to the fused feature vector; according to the method, the long-term stable operation capability, the performance repeatability and the reliability in a standardized application scene of the high-throughput cell sorting system can be effectively improved, and the complexity and time consumption of system calibration and troubleshooting are reduced.

The invention discloses a portable multi-stage cell sorting device and a sorting method thereof, and belongs to the technical field of cell sorting. A pull plate and a rotating rod drive a deflection seat to rotate, so that the deflection seat pulls a sliding plate through a hinge rod, and the sliding plate drives a clamping strip to be separated from a clamping groove, so that a centrifugal rotor is conveniently detached; the centrifugal rotor is connected to the top of the butt joint base in a sleeving mode, after the clamping base is completely clamped into the rectangular groove, the deflection base is driven to rotate through the torsion effect of the torsion spring, the deflection base pushes the sliding plate to move through the hinge rod, the sliding plate drives the clamping strip to be clamped into the clamping groove, and the purpose of locking the centrifugal rotor and the butt joint base is achieved. A traditional mode of adopting thread locking or bolt fixing is abandoned, auxiliary calibration is carried out on the centrifugal rotor through the butt joint base, the central axis of the centrifugal rotor and the central axis of the butt joint base are concentric, the situation that dynamic balance is affected by deviation during installation is prevented, the centrifugal rotor is disassembled and assembled through a clamping structure, and operation convenience and maintenance efficiency are improved.

The application discloses a micro-fluidic chip for sorting sub-micron scale cells, which is composed of three components of an upper cover plate, a cell sorting micro-channel and a lower bottom plate by layer bonding. The upper cover plate is provided with an outside inlet, an inside inlet, an outside outlet and an inside outlet. The cell sorting micro-channel is composed of three regions of an inlet structure adjusting region, a spiral channel region and an outlet region. The lower bottom plate is provided with a convex wedge-shaped structure, and three kinds of inlet structures of an outside-narrow-inside-wide shape, an outside-inside-equivalent shape and an outside-wide-inside-narrow shape can be obtained by adjusting the position of the wedge-shaped structure. The sub-micron scale cells are sorted by the cooperation of the cell sorting micro-channel and the wedge-shaped structure. The spiral micro-channel is introduced, the cell transverse migration starting point is controlled by adjusting the spiral channel inlet structure and cooperating with suitable flow conditions, and the high-precision sorting of the sub-micron scale cells is finally completed. The cell sorting diversity and adaptability are improved by flexibly adjusting according to specific sorting requirements.

The utility model relates to the technical field of magnetic frames, and discloses a cell sorting magnetic frame which comprises a base, the top of the base is fixedly connected with a fixing plate, the top of the fixing plate is provided with a top plate, the top of the top plate is provided with a placing groove, the top of the base is provided with a positioning groove, and the side face of the fixing plate is provided with an installation groove. Mounting grooves are formed in the side faces of the fixing plates, magnetic strips are mounted in the mounting grooves, connecting grooves are formed in the side faces of the fixing plates, mounting plates are mounted in the connecting grooves, and bolts are in threaded connection with the surfaces of the mounting plates. According to the cell sorting magnetic frame, the base and the fixing plate are used in cooperation, meanwhile, the mounting grooves are utilized, so that magnetic strips are conveniently mounted in the mounting grooves, when the magnetic strips are fixed, the mounting plates can be mounted in the connecting grooves, meanwhile, bolts are rotated, the positions of the mounting plates can be fixed, and therefore the magnetic strips can be fixed.

Disclosed herein is a method for training a network to detect and classify abnormal pathologies in images if cells. Specifically, the network uses a deep framework optimally trained to detect and classify abnormal cervical cells in pap smear images.

The invention discloses a method for high-throughput screening of high-activity lactic acid bacteria based on multi-index fusion and application, and belongs to the field of lactic acid bacteria screening, the method comprises the steps of bacterial cell enrichment, single-cell suspension preparation, physiological status and activity evaluation of cells, flow cytometry analysis, cell sorting and strain identification; in the step of enrichment of the bacterial cells, a sample is activated and enriched by using an improved liquid culture medium and an improved solid culture medium, and activated and enriched bacterial suspension is obtained. The invention provides the method for detecting the activity of the fermentation product and the lactic acid bacteria stored for a long time, the method is convenient and rapid, the screening time is greatly shortened, and the screening efficiency is improved.

One approach to the exploration method includes calculating a TAD score representing transcriptional activity for each TAD present in the genome and selecting TADs based on the TAD scores. Another approach includes analyzing a genome obtained from cells expressing a foreign gene and selecting the boundaries of regions containing the foreign gene. Cell preparation involves inserting the target gene into the regions identified by the exploration method. Cell sorting involves calculating a TAD score representing transcriptional activity for each TAD present in the genome, selecting TADs based on the TAD scores, and selecting cells containing the target gene in the selected TADs. The cells are derived from Chinese hamsters and are cells in which the target gene has been inserted into at least one specified region. The method for manufacturing the cell product includes culturing the cells to express the target gene.

A cellular mechanical property measurement and sorting method comprises: cells to be sorted being arranged in a straight line after passing through a standing wave acoustic field generated by a first interdigital transducer; the cells flowing through a focused acoustic field generated by a second interdigital transducer, and the focused acoustic field generating a radiation force on the cells, such that the cells are deformed; performing calculation according to information of the radiation force and deformation, so as to obtain elastic moduli of the cells; determining the positions of the cells, and selectively driving, with designed delay time, interdigital transducers in a cell sorting unit according to the moving speeds of the cells and the distances between the cells and the sorting unit, and triggering, according to the elastic moduli, a corresponding interdigital transducer to generate a planar acoustic field, so as to sort the cells.

The present application relates to a kind of nanometer magnetic beads for cell sorting and its preparation method.The preparation method of nanometer magnetic beads of the present application includes the following steps: (1) under ice bath condition, prepare aqueous solution containing trivalent iron and divalent iron;(2) using alkaline substance to adjust the pH value of the aqueous solution containing trivalent iron and divalent iron to 10-12;(3) reaction is obtained under 65-90 DEG C aqueous solution containing the nanometer magnetic beads.The preparation method of the present application can control the median particle size of nanometer magnetic beads in 50-200nm, and good particle size uniformity, while guaranteeing the sorting efficiency of cell, the function of target cell is little, and biocompatibility is good, has good application prospect in the field of biological medicine.In addition, the preparation method of the present application is simple and easy to operate, and is suitable for large-scale industrial production.

The utility model discloses a soil sample cell sorting pretreatment device, which belongs to the technical field of soil sample pretreatment and comprises an outer cylinder and an inner cylinder, the inner cylinder is rotatably mounted in the outer cylinder, a top cover is fixedly mounted at the top of the outer cylinder, a first motor is fixedly mounted at the top of the top cover, and a second motor is fixedly mounted at the bottom of the inner cylinder. The output end of the first motor is fixedly connected with a first stirring rod, a second motor is fixedly mounted on one side of the outer cylinder, the output end of the second motor is fixedly connected with a bevel gear, and the outer surface of the inner cylinder is fixedly sleeved with a bevel gear ring meshed with the bevel gear; three annularly-distributed second stirring rods are rotatably mounted on the inner bottom wall of the inner barrel, driven gears are fixedly mounted at the upper ends of the second stirring rods, and a fixed gear ring is fixedly connected to the bottom of the top cover. A second motor, a bevel gear, a bevel gear ring, a second stirring rod, a driven gear and a fixed gear ring are arranged, so that the soil sample can be fully stirred, and the mixing efficiency is improved.

The utility model relates to the technical field of cell sorting and detection, in particular to a micro-fluidic chip. The micro-fluidic chip comprises a substrate, a micro-fluidic chip, a micro-fluidic chip and a micro-fluidic chip, wherein a metal film layer is arranged on the substrate; the cover plate is at least provided with a micro-channel; the intermediate bonding layer is located between the substrate and the cover plate and covers the metal film layer; wherein the cover plate is a PDMS (Polydimethylsiloxane) prepolymer cover plate; the intermediate bonding layer is a silicon dioxide layer. According to the utility model, the problem that the PDMS prepolymer cover plate and the metal film layer substrate are difficult to directly bond can be solved.

The application discloses a cell sorting method and system based on an image acoustic flow control cell sorting model and a computer readable storage medium, the image acoustic flow control cell sorting model comprises a cell image recognition module and a cell ejection module, and the cell sorting method comprises the following steps: acquiring an original cell image set and a predetermined sample cell image set; inputting the sample cell image set into the cell image recognition module for feature extraction to obtain image category information of the cell image; inputting the image category information and the sample image set into the image acoustic flow control cell sorting model for training; inputting the original cell image into the trained image acoustic flow control cell sorting model for image prediction to determine a target image corresponding to the image category; and ejecting a cell corresponding to the target image to a preset collection area based on the cell ejection module. In the embodiment of the application, the automatic classification of cells can be performed by using a label-free method, so that the purification and collection of target cells are realized.

The invention relates to a cell sorting system and method based on a micro-fluidic chip and micro-nano control, and relates to the technical field of cell sorting, the system comprises the micro-fluidic chip composed of a top layer, a middle layer and a bottom layer which are attached in sequence; a main channel and a diversion channel are arranged at the boundary of the top layer and the middle layer, the diversion channel is communicated with the side wall of the main channel, and the middle layer is provided with a sampling hole located in the bottom wall of the main channel; a sample channel is arranged at the boundary of the middle layer and the bottom layer; the sample introduction hole only allows single cells to pass through in sequence; under the limitation of the size of the sample injection hole, a sample flow in the sample channel enters the main channel to form a narrow sample flow. When the single cell data acquisition time of the cell detection module is less than the time difference of two adjacent cells flowing through the capture point of the micro-nano control module in the narrow sample flow, high-throughput cell detection can be carried out in the narrow sample flow.

This specification describes a framework that includes one or more feature extractors (models) and a cluster component for clustering. The framework supports (1) offline, image-based, unsupervised clustering, which replaces time-consuming manual gating, and (2) online, image-based single-cell sorting. During training, one or more cell image datasets, with or without ground truth, are used to train the one or more feature extractors. The feature extractors are based on neural networks containing several convolutional layers. Once trained, the feature extractors are used to extract cell image features for unsupervised cell clustering and sorting. Additionally, after the feature extractors are trained, they can be further refined using additional datasets.

Aspects of the present disclosure include methods for adjusting a particle classification index in response to one or more saturated data signals based on detected light from particles in a fluid stream. Methods according to certain embodiments include detecting light from particles in a fluid stream, generating a plurality of data signals based on the detected light, identifying one or more saturated data signals, generating a saturation signal index corresponding to the identified saturated data signals, and applying the saturation signal index to a particle classification index to generate an adjusted particle classification index. In some embodiments, the methods include determining one or more parameters of a particle (e.g., for a particle sorting decision) by calculating an adjusted spectral unmixing matrix for fluorescence of the particle that excludes the one or more saturated data signals. Systems and integrated circuit devices (e.g., field programmable gate arrays) for implementing the subject methods are also provided. Non-transitory computer readable storage media are also described.

The present invention discloses a magnetically driven portable multi-cell separation device. According to one embodiment of the present invention, the magnetically driven portable multi-cell separation device includes: A microscope capable of nano-scale observation, A chip unit, positioned below the microscope, enabling selective cell sorting and capture, A magnetic field generator, which is adjacent to the chip unit and can control the magnetic field via a magnetic member, and A control unit, which controls the operation of the imaging unit, the chip unit, and the magnetic field generator.

This invention discloses an APLN monoclonal antibody, its preparation method, and its applications. The APLN monoclonal antibody includes a light chain variable region and a heavy chain variable region. The amino acid sequence of the light chain variable region is shown in SEQ ID No:1, and the amino acid sequence of the heavy chain variable region is shown in SEQ ID No:2. The APLN monoclonal antibody is obtained by immunizing mice with APLN protein, followed by single-cell B-cell sorting and single-cell PCR. Utilizing advanced single-cell B-cell cloning technology combined with single-cell PCR, a novel APLN monoclonal antibody has been developed. The obtained antibody sequence is the result of natural affinity maturation screening by the in vivo immune system, requiring no additional humanization or in vitro affinity maturation steps. It retains the natural high affinity and specificity, and can directly obtain all-natural antibodies with high affinity and high specificity.