355 results about "Flow cytometry" patented technology

Aspects of the present disclosure include methods for identifying measurement uncertainty associated with light detected from a sample. Methods according to certain embodiments include introducing a sample into a flow cytometer, flowing the introduced sample in a flow stream, irradiating the sample in the flow stream with a light source, detecting light from particles in the sample flowing in the flow stream and, identifying measurement uncertainty associated with the detected light. In some embodiments, measurement uncertainty is identified corresponding to individual particles in the sample. In certain embodiments, measurement uncertainty is identified for individual parameters of detected light for particles in the sample. Methods according to some embodiments further comprise generating a quality score for each particle based on the measurement uncertainty for each particle. Systems, integrated circuit devices (e.g., a field programmable gate array) and non-transitory computer readable storage mediums for practicing the subject methods are also provided.

The invention belongs to the technical field of flow cytometers, and discloses a flow velocity calibration method of a flow cytometer and related equipment, an image sequence of sheath liquid drops is obtained under different sheath liquid pressures, an effective image group is screened out by using a multi-dimensional feature association confirmation method, and then the flow velocity is calculated; on the basis, carrying out screening and characteristic calculation on the flow velocity data, and generating mapping relation information between flow velocity characteristics and sheath fluid pressure; and finally, based on a composite control strategy combining feedforward and feedback, sheath fluid pressure control is carried out in combination with the mapping relation information, so that the flow velocity of the sheath fluid drops is stabilized at the target flow velocity. The method effectively solves the problems of inaccurate flow velocity control and poor stability caused by a complex nonlinear relationship between the sheath fluid pressure and the actual flow velocity of a flow cytometer in the prior art.

The utility model relates to a micro-fluidic flow cytometry fluorescence detector which comprises an excitation light source optical fiber, a light beam shaping module, a focusing and fluorescence collecting module and a micro-fluidic chip which are sequentially arranged along the excitation light path direction of the excitation light source optical fiber, and a multi-channel silicon photomultiplier tube is arranged above the micro-fluidic chip. A plurality of groups of detection channels for receiving fluorescence signals of different wave bands are arranged in the multichannel silicon photomultiplier tube at intervals up and down, and a dichroscope, a band-pass filter and a lens group are sequentially arranged in each group of detection channel along a light path and are gathered on a receiving chip; the micro-fluidic flow cytometry fluorescence detector can efficiently and accurately complete multi-wavelength fluorescence excitation and signal collection, and is suitable for multi-channel and multi-wavelength fluorescence detection requirements in a micro-fluidic chip.

The present invention relates to a system and a method for measuring and analyzing minimal residual disease (MRD) in pediatric B-cell precursor acute lymphoblastic leukemia (B-ALL) using multiparameter flow cytometry (MPFC). The invention finds application in clinical diagnostics and hematology-oncology for quantifying MRD in B-ALL patients with high sensitivity and specificity, needed for risk stratification, monitoring treatment response, and informing therapeutic decisions. The system comprises interconnected subsystems including an acquisition subsystem with an MPFC instrument, a control and file generation subsystem, and an analytical subsystem. The analytical subsystem incorporates modules for sequential data reduction, automated data cleaning, automated unsupervised data clustering, and interactive cluster analysis. Key advantages include high MRD detection sensitivity (e.g., 10⁻⁵ or 0.001%) and high specificity, without reliance on reference samples or supervised machine learning models, making it applicable in laboratories with different measuring equipment and using different panels of antibodies for identification of leukemic cells.

The invention provides a polypeptide for activating cellular immunity in chronic hepatitis B and application of the polypeptide, and belongs to the technical field of biological medicine. A polypeptide library is designed and synthesized on the basis of a preS1 structural domain for coding HBV large HBsAg, a full-length core protein Core, a polymerase protein fragment rich in T cell epitopes and an mRNA-PreS1CPX holoantigen sequence (as shown in SEQ ID NO.1) of full-length X protein HBX, and peptide fragments capable of activating T cell immunity are screened by utilizing ELISPOT and flow cytometry. Experimental results show that the polypeptide sequences as shown in SEQ ID NO.2-15 can promote HBV antigen specific immune response by stimulating CD8 + T lymphocytes to secrete IFN-gamma, so that immune activation treatment of hepatitis B is realized.

The invention discloses a regulation mechanism research method of depleted precursor CD8T cells, and relates to the field of immunology and molecular biology. Comprising the following steps: detecting the expression level of the ARHGAP9 gene in a chronic virus infection model; constructing a T cell specific ARHGAP9 gene knockout animal model; the influence of ARHGAP9 deletion on the frequency, phenotype and function of the Tpex cell is analyzed; a single cell transcriptome sequencing technology reveals that ARHGAP9 regulates and controls a downstream signal channel of a Tpex cell. By constructing a chronic virus infection model, detecting the expression dynamic state of the ARHGAP9 gene, knocking out an animal model by utilizing T cell specificity ARHGAP9, and combining flow cytometry, qPCR, single cell transcriptome sequencing and other technologies, the invention discloses the effect of the ARHGAP9 as a novel immune checkpoint molecule, and provides a theoretical basis for developing Tpex cell targeting immunotherapy.

The application discloses a flow cytometry sample processing system and belongs to the technical field of automatic processing of biological samples. The flow cytometry sample processing system comprises a machine table, a flow tube block placing structure, a transfer mechanical arm, a shaking module and a centrifuge. The flow tube block placing structure is arranged on the machine table, a flow tube block is arranged on the flow tube block placing structure, a flow tube is arranged on the flow tube block, the transfer mechanical arm provided with a clamping jaw can integrally transfer the flow tube block provided with a clamping hole to the shaking module and the centrifuge, and the whole process automation is truly realized, and the sample centrifugal processing efficiency is improved.

The present application relates to the technical field of biotechnology, in particular to a programmed death receptor ligand 2 monoclonal antibody and application thereof. The present application provides a PD-L2 monoclonal antibody, amino acid sequences of three CDR regions of a heavy chain thereof; amino acid sequences of three CDR regions of a light chain thereof; or amino acid sequences obtained by substitution, deletion or addition of one or more amino acids of the amino acid sequences, or amino acid sequences functionally identical or similar to the amino acid sequences; or amino acid sequences having at least 80% homology with the sequences. The present application widens the possibility of antibody therapy for blocking PD1 / PD1 ligand, and compared with existing PD-L2 monoclonal antibodies, the 8E3 antibody has unique amino acid sequences and CDR region sequences, high affinity and high specificity. The 8E3 antibody can be used in ELISA, Western blot and flow cytometry detection, and has wide application.

The invention discloses a culture method for inducing high expression of umbilical cord mesenchymal stem cells (LRP1) and application, and belongs to the technical field of biological medicines. The technical problem to be solved is to provide a standardized culture method for directionally inducing a specific functional phenotype cell population, so that efficient and accurate induction of LRP1 high-expression UCMSCs is realized, and clinical adaptation is met. According to the key point of the technical scheme, the culture method for inducing the LRP1 high-expression umbilical cord mesenchymal stem cells comprises the following steps: inoculating umbilical cord mesenchymal stem cells into a culture dish at the inoculation density of 5.5 * 10 < 3 > / cm < 2 > to 3.6 * 10 < 4 > / cm < 2 >, and culturing for 24-72 hours by using a special mesenchymal stem cell culture medium without adding a supplement, so that the acquisition efficiency of the LRP1 high-expression UCMSCs is improved, and the yield of the LRP1 high-expression UCMSCs is increased. Antibody labeling and flow sorting are not needed, so that the operation steps are simplified, and the complexity and cost of the culture process are reduced.

Provided are methods for flow cytometry analysis, including the setting and use of static gating thresholds separating positive fluorescence from negative fluorescence for each fluorescence channel in the assay.

This invention provides an automatic sample allocation method for flow cytometry detection, belonging to the field of flow cytometry cell detection technology. The method includes: acquiring basic sample information; pre-analyzing the sample using a microfluidic chip to obtain sample density, viscosity, and ion concentration; measuring the microchannel diameter and length of the microfluidic chip using a laser ranging system and recording the microchannel material properties; setting a target allocation volume, starting a high-precision injection pump system for pre-injection, and obtaining the initial allocation rate by measuring the flow rate using a flow sensor; calculating the corrected allocation rate using fluid dynamics compensation equations; controlling the high-precision injection pump system to allocate the sample according to the corrected allocation rate; monitoring the sample liquid level in real time using an ultrasonic sensor and synchronously acquiring the real-time allocation rate; and performing sample diversion through the microfluidic chip to achieve multi-channel parallel allocation.

The invention relates to a binding protein B capable of binding with klebsiella pneumoniae and application of the binding protein B, and belongs to the technical field of molecular biology and protein engineering. The binding protein B provided by the invention can be specifically bound with an extramembrane part of a specific protein P24017. OMPAKLEPN of klebsiella pneumoniae, and experiments such as bimolecular fluorescence complementation and flow cytometry prove that the binding protein B has good recognition and stable binding capacity on the klebsiella pneumoniae, can be used for identifying and screening the klebsiella pneumoniae, and has good application prospects. And a key tool is provided for targeted drug development aiming at the bacterium.

The invention discloses an application of Northeast tiger interleukin-15 in preparation of an antiviral drug. According to the invention, IL-2 family genes (IL-2, IL-4, IL-9, IL-15 and IL-21) and IL-13 genes of the northeast tiger are cloned and successfully expressed. Functional experiments show that all cytokines can promote lymphocyte proliferation, F81 cells are pretreated with IL-15 and then exposed to feline panleucopenia virus (FPV) derived from northeast tiger, and different exposure time is set for subsequent experimental research. The consistency of flow cytometry, protein expression analysis and ROS staining results shows that IL-15 can significantly weaken oxidative stress and apoptosis induced by FPV infection. In conclusion, the molecular and functional characteristics of the northeast tiger IL-2 cell factor family are clarified, and IL-15 is highlighted as a cell factor with a potential antiviral protection effect. The results provide technical means for cytokine-based immunological research and potential treatment strategy development to support health and protection of northeast tigers in the future.

The application discloses a method for establishing a mouse model of MHC haplotype compatible allogeneic hematopoietic cell transplantation, and belongs to the technical field of animal experiment model construction. The technical problem to be solved by the application is how to construct a mouse model which can be used to study the immune reconstruction mechanism and treatment effect evaluation of MHC haplotype compatibility and MHC full compatibility in bone marrow cell transplantation recipient mice. To solve the technical problem, the application provides a method for constructing a mouse model of MHC haplotype compatible allogeneic hematopoietic cell transplantation, which comprises transplanting bone marrow cells of a donor mouse into a recipient mouse, and the MHC haplotype of the donor mouse and the recipient mouse is compatible. The application sets a modeling condition, and the model establishment condition is evaluated by survival condition, graft-versus-host disease and rejection reaction observation, flow cytometry and histopathological section, so as to prove that the model is reliable and stable.

The invention belongs to the field of immunotoxicology, and particularly relates to a method for immunotyping bone marrow cells by flow cytometry. The flow cytometry immune typing method for bone marrow cells comprises the following steps: collecting macaca fascicularis bone marrow fluid, and preparing the macaca fascicularis bone marrow fluid into single-cell suspension; dyeing the obtained cells by using an antibody combination containing a plurality of fluorescent labels; then, a flow cytometer is used for collecting cell fluorescence signals, a standardized gate setting strategy is executed, and based on collected data, the percentage of each cell subset in CD45 + living white blood cells is calculated to complete immune typing. According to the flow cytometry immune typing method for the bone marrow cells, the problems of low efficiency, low repeatability, low sensitivity and poor specificity of related technologies are solved, and more accurate and reliable data support is provided for early discovery, risk assessment and mechanism research of drug immunotoxicity.

The invention provides a respiratory tract infection pathogen antibody spectrum detection method and a respiratory tract infection pathogen antibody spectrum detection system, and relates to the technical field of biomass energy industries. If yes, performing gain dynamic adjustment based on the instrument stability dynamic evaluation result and the data quality parameters, performing antibody spectrum detection by using the instrument subjected to gain dynamic adjustment, and if the instrument stability dynamic evaluation result is a normal instrument, directly performing antibody spectrum detection by using the instrument; performing flow velocity rationality dynamic evaluation based on the flux efficiency demand and the sample core parameter, and executing liquid flow dynamic adjustment based on the liquid flow quality control parameter according to a flow velocity rationality dynamic evaluation result; and dynamically generating the antibody spectrum of the respiratory tract infection pathogen based on the microsphere coding information and the fluorescence intensity signal output by the flow cytometry after the liquid flow dynamic regulation.

The application belongs to the technical field of flow cytometry signal processing, and particularly relates to a flow cytometer signal background dynamic acquisition method based on window sliding screening, which comprises the following steps: S1. selecting a signal; S2. setting a screening parameter: setting an initial screening window size as N units, and setting an initial reasonable standard deviation of the initial screening condition signal as σ0; S3. finding a first reasonable window: slidingly finding with the screening window with a size of N units, calculating a signal value mean μ1 and a signal value standard deviation σ1 of the signal Vt in the screening window; when the standard deviation σ1 meets the condition of being less than the reasonable standard deviation σ0, the window is the first reasonable window; S4. dynamic adjustment of the screening condition: after the nth reasonable window is found, the screening condition is updated as μ n - 3σ n ≤ the signal value of Vt ≤ μ n + 3σ n , and the (n+1)th reasonable window is found with the condition, the signal value of the signal Vt in the (n+1)th reasonable window meets the condition of μ n - 3σ n ≤ the signal value of Vt ≤ μ n + 3σ n ; and S6. real-time extraction of a background value.

A sample preparation instrument includes a pod-arm module and a reagent module that includes a housing defining an interior cavity and a lid attached to the housing. The sample preparation instrument receives an instruction to prepare a cocktail having predefined volumes of reagents, and automatically generates the reagent cocktail by controlling the pod-arm module to move one or more probes to aspirate the predefined volumes of the reagents and to dispense the predefined volumes of the reagents into at least one designated container while maintaining a refrigerated and dark environment inside the reagent module.

The invention provides a multicolor flow-based synaptic heterogeneity analysis device and analysis method and application thereof, the analysis device comprises: (1) a scattering-resolution pretreatment module: adopting a BD FACS fusion double laser platform, firstly positioning an FSC-SSC coordinate by using a standard 0.5 m polystyrene microsphere, and then loading a synaptic body; the adopted sheath fluid comprises HPMC (Hydroxymethyl Methacrylate)-3cp and 1 * PBS (Phosphate Buffer Solution) without Ca < 2 + > / Mg < 2 + >; an adopted carrier buffer solution comprises EDTA (Ethylene Diamine Tetraacetic Acid), a 1 * protease inhibitor mixture and 1 * PBS (Phosphate Buffer Solution) without Ca < 2 + > / Mg < 2 + >; and (2) a multi-color flow type synchronous marking-sorting module which is used for detecting two kinds of code fluorescent marks in parallel on double laser platforms of 488 nm and 561 nm, eliminating cross color by a logic gate-double exponential decay compensation algorithm, and realizing transmitter and function sorting.

A robotic system for automated sample preparation is provided. A system of interest includes: a plurality of sample processing modules; a plurality of robotic components integrated with the sample processing module; a processor including a memory operably coupled to the processor, where the memory includes instructions stored thereon that, when executed by the processor, cause the processor to control the sample processing module and the robotic assembly, to: operate the sample processing module and robotic assembly to prepare a plurality of samples for flow cytometry analysis; and an operable connection between the processor, the sample processing module, and the plurality of robotic components. Systems according to certain embodiments also include a flow cytometer, where the plurality of robotic components are further integrated with the flow cytometer, where the memory further includes instructions stored thereon, the instructions, when executed by the processor, cause the processor to control the sample processing module, the robotic assembly, and the flow cytometer to: load each prepared sample of the plurality of samples into the flow cytometer; and operating the flow cytometer to analyze each prepared sample of the plurality of samples, wherein the operable connection operably connects the processor, the sample processing module, the flow cytometer, and the plurality of robotic components together. Methods of use and configurations or designs of the system are also provided.

Systems and methods for analyzing flow cytometry particles. A flow cytometry system is configured to direct a fluid stream of particles through an interrogation location, and includes a laser configured to emit light toward the interrogation location to produce light signals from the particles, and one or more detectors configured convert the light signals to waveform data. The flow cytometry system includes a waveform acquisition device to continuously digitize the waveform data, and a graphics processing unit configured to apply one or more adjustable threshold voltages to the digitized waveform data to extract event data for the particles.

The invention relates to a quality control substance for flow cytometry HLA-B27 analysis as well as a preparation method and application of the quality control substance. The quality control substance comprises a negative quality control substance and a positive quality control substance; the negative quality control substance comprises human lymphocyte negative quality control substance simulation particles and human peripheral blood cells or simulation particles; an HLA-B27 antigen is not expressed on the surface of the human lymphocyte negative quality control substance mimic particle; the positive quality control substance comprises human lymphocyte positive quality control substance simulation particles and human peripheral blood cells or simulation particles; an HLA-B27 antigen is expressed on the surface of the human lymphocyte positive quality control substance simulation particle; cD3 antigens and CD45 antigens are expressed on the surfaces of the human lymphocyte positive quality control substance simulation particles and the human lymphocyte negative quality control substance simulation particles. The technical problem to be solved is how to provide a quality control substance which is good in stability and can be applied to flow cytometry HLA-B27 analysis.

The invention provides methods for classifying fluorescent flow cytometry data. In some cases, the methods include processing the flow cytometry data with a supervised algorithm configured to cluster the fluorescent flow cytometry data into different populations based on the relationship of data points to associated thresholds. In embodiments, the methods include calculating and combining spill-over diffusion coefficients in a spill-over diffusion matrix to determine the extent of spill-over diffusion. In some embodiments, the fluorescent flow cytometry data populations are adjusted to reduce the extent of spill-over diffusion. In embodiments, the spill-over diffusion adjusted populations are partitioned after evaluating potential partitions relative to the thresholds. In embodiments, the partitioned fluorescent flow cytometry data populations are classified (i.e., phenotyped) according to a hierarchy. The invention also provides systems and computer readable media for classifying fluorescent flow cytometry data.

Provided are a set of fully human TSH receptor (TSHR)-blocking monoclonal antibodies targeting a CHK36 homolog cluster, a preparation method therefor, and an application thereof. The method comprises the following steps: using flow cytometry to sort plasma cells and single memory B cells that specifically recognize TSHR in peripheral blood of a patient with a high titer of TSH-blocking antibodies (TBAb), performing in vitro cloning of antibody light and heavy chains and performing recombinant expression, and using hTSHR-CHO cells to perform screening and validation of antibody properties to obtain a blocking monoclonal antibody that specifically targets human TSHR. The fully human TSH receptor-blocking monoclonal antibodies specifically bind to a TSHR, and effectively block signal transduction after a TSH binds to a receptor, inhibit the synthetic secretion of thyroid hormones, and significantly reduce and inhibit TSHR expression and fibrosis in orbital fibroblasts of effector cells associated with thyroid-related ocular diseases. The fully human TSH receptor-blocking monoclonal antibodies have broad application prospects in the treatment of Graves' disease (GD) and other diseases caused by hyperthyroidism, such as thyroid eye disease (TED).

Disclosed is an efficient and non-genetically modified iPSC-induced, industrialized single clone selection platform, and a use. The platform can efficiently perform reprogramming, and only requires the use of a minimal number of reprogramming factors (OCT4, SOX2, E6, E7). During the single clone separation stage of the present invention, SSEA4 / TRA-1-60 is used as a screening marker, and a large number of single cell clones are obtained by means of flow cytometry. The platform described in the present invention has advantages such as high reprogramming efficiency, high safety, easy operation, and large-scale production.

The invention provides a detection method for reducing the off-target risk of a targeting drug, and relates to the technical field of immunodetection. The detection method comprises the following steps: screening the optimum working concentration of a test substance, screening the human plasma membrane protein having a binding risk with the test substance, and screening out a target protein having a specific binding relationship with the test substance; according to the detection method provided by the invention, the expression of the fluorescent protein can indicate the expression condition of the human plasma membrane protein, positive control, quality control control and negative control are set, and the target protein having a specific binding relationship with a test substance is obtained through chemiluminescence detection and flow cytometry screening, so that the risk of a false negative result is reduced, and the detection accuracy is improved. And the accuracy of predicting the off-target risk of the targeted drug is improved.

The present application adopts the way of plant explant callus culture, adds carbon source containing fluorescent group in the culture medium, induces callus with fluorescent group, thin cell wall, easy to be single enzymolysis, uniform cell morphology and size, the cell after single enzymolysis is detected by flow cytometry, the purity and activity of the prepared leaf single cell are 98.5% and 99.95 respectively, the agglomeration rate is only 1.5%, the cell purity and activity are increased by 32.6% and 21.05 respectively compared with the traditional technical scheme, and the agglomeration rate is reduced by 31.0%; the rice seed is used as the explant, and the seed single cell with cell purity and activity of 99.5% and 100% is obtained, which breaks through the limitation of traditional enzymolysis method that cannot prepare seed single cell; in addition, through the common induction of safflower leaf and rice seed, the cell purity and activity prepared are as high as 99.98% and 100.0, which overcomes the limitation of single cell preparation caused by interspecific heterogeneity.

This invention discloses an absolute quantification method for pathogens and drug resistance genes in atmospheric bioaerosols and its application. Addressing the bottleneck of achieving absolute quantification of low-concentration samples while maintaining cell integrity and high-concentration nucleic acid extraction, this invention constructs a collection carrier by immobilizing a hydrophilic mixed cellulose ester microporous membrane on a technical agar plate. This is combined with a flow rate ≤30 L / min impactor sampler, ensuring structural integrity while capturing microorganisms, laying the foundation for flow cytometry quantification. This front-end collection system is deeply coupled with a dedicated cascade lysis and concentration process, enabling the acquisition of high-quality DNA suitable for metagenomic library construction and relative abundance analysis even under short-term, low-flow-rate collection. Simultaneously, the total microbial count is absolutely quantified using flow cytometry on the collected samples, and this result is fused with the relative abundance results. Through synergistic optimization of the entire chain, quantitative analysis of species-level pathogens and drug resistance genes in atmospheric bioaerosols is ultimately achieved.

The application discloses a kit for predicting the curative effect of immunotherapy on prostate cancer patients, which comprises reagent A, reagent B, reagent C, reagent D, reagent E and reagent F; the reagent A is a PBS buffer solution; the reagent B is a lymphocyte separation medium; the reagent C is a red blood cell lysis solution; the reagent D is a cell surface CD3 flow antibody; the reagent E is a cell surface CD19 flow antibody; and the reagent F is a 1% polyformaldehyde solution in terms of mass percentage concentration. The application further discloses an application method of the kit, which comprises the following steps: collecting venous blood of prostate cancer patients; extracting peripheral blood mononuclear cells (PBMC) in the venous blood of the prostate cancer patients; incubating the flow antibody; and performing flow cytometry analysis. The kit and the application thereof are simple and fast, non-invasive, repeatable, low in cost and short in detection time.

Inventors have first investigated the impact of PIK3CA inhibition in NZBWF1 / J mice a model of lymphoproliferative disorders. They randomly assigned 30 females aged of 24 weeks to receive either vehicle (n=15) or alpelisib (n=15) during 4 weeks. At the time of sacrifice, alpelisib treated mice demonstrated significantly reduced spleen size. Flow cytometry analysis revealed that B cells were significantly reduced in alpelisib treated mice and CD8 cells count corrected. They then decided to explore the relevance of alpelisib in MRL / MpJ-Faslpr / J mice (referred here as MRL-lpr), another mouse model of lymphoproliferative disorder. These mice with homozygous Fas mutation usually develop severe lymphadenoproliferation. At the time of sacrifice. MRL-lpr mice treated with alpelisib demonstrated a reduction on their spleen and lymph node sizes. Flow cytometry analysis showed correction of B cells. T cells and other immune cells in peripheral blood mononuclear cells (PBMC), lymph nodes and spleen. The invention relates to a method for treating lymphoproliferative disorder in a subject in need thereof comprising a step of administering the subject with a therapeutically effective amount of a PIK3CA inhibitor.