775 results about "Flow cytometry" patented technology

A sample preparation instrument with an integrated device for estimating a concentration of white blood cells in a specimen is described. The sample preparation instrument receives a specimen for which a sample is to be prepared for analysis. The integrated device can implement an optical method, an electrical resistance method, or a flow cytometry method, for example, to estimate the white blood cell concentration in the specimen. For some types of analysis, a sample volume of the specimen is dependent on the white blood cell concentration. Therefore, the sample preparation instrument can automatically determine and adjust the sample volume of the specimen based on the estimated white blood cell concentration prior to adding additional reagents, such as labeling reagents and / or lytic reagents, to prepare the sample.

The invention provides a tumor detection data analysis method based on a peripheral blood immune marker, and belongs to the technical field of immunodetection.The tumor detection data analysis method comprises the steps that firstly, a peripheral blood sample of a patient is collected, and plasma and cell components are separated; analyzing various immune cell phenotypes by using flow cytometry; determining the concentration of key cell factors; analyzing the molecular expression level of the immune checkpoint; establishing an immune cell subset proportion empirical function; analyzing a cell factor expression mode by using an immune lineage transcriptome analysis model; establishing a tumor immune microenvironment score by combining the immune cell subset proportion and a model output result; the accuracy of the scoring system is verified through clinical follow-up visit and iconography examination; a standardized report template is formulated to provide a tumor early detection basis. Wherein the immune lineage transcriptome analytical model adopts a multi-modal attention fusion network architecture, three different immune data streams can be processed and integrated at the same time, and accurate evaluation of tumor-related immune states is realized.

The invention provides a sample pretreatment automation method for flow cytometry, and belongs to the technical field of sample pretreatment for flow cytometry. A biological sample to be detected is collected and placed in a collection tube containing an EDTA-K2 anticoagulant for pretreatment; an automatic pipetting system is used for carrying out incubation reaction on an anticoagulant whole blood sample and an intelligent matching fluorescence labeling antibody mixed solution, and a double-layer game optimization verification system is used for verifying the rationality of an antibody adding proportion and reaction condition parameters; carrying out fluorescence characteristic readability index prediction on the incubated cell suspension by adopting an intelligent dilution parameter regulator model, dynamically adjusting hemolysis parameters and suspension conditions, and adding 1 * hemolysin working solution by using an automatic liquid adding device to carry out hemolysis treatment so as to finish pretreatment; the technical problems that in the pretreatment process of flow cytometry detection samples, parameters are often based on artificial experience, and the reproducibility and standardization degree of detection results are insufficient are solved.

The invention discloses a monoclonal antibody BY017 combined with African swine fever virus P30 protein and application of the monoclonal antibody BY017. The invention provides an IgG antibody which is composed of a light chain and a heavy chain. CDR1, CDR2 and CDR3 in the heavy chain variable region are sequentially shown as the 47 to 54 , the 72 to 79 and the 118 to 126 in SEQ ID NO: 1; cDR1, CDR2 and CDR3 in a light chain variable region are sequentially as shown in the 49 to 60 , the 78 to 80 and the 117 to 124 in SEQ ID NO: 3. The invention also protects the application of any one of the IgG antibodies in preparation of products. The product has the functions that the product is used for being combined with an African swine fever virus P30 protein fragment; the antibody is used for binding African swine fever virus P30 protein; the antibody is used for binding African swine fever virus. The IgG antibody provided by the invention can be used for multiple purposes of western blot, immunohistochemistry, flow cytometry, ELISA (Enzyme-Linked Immunosorbent Assay) and the like, and can be used as an important tool for ASFV detection and structural analysis of P30 protein of ASFV detection.

The invention relates to the technical field of antibodies, and provides an anti-CD45 antibody for detecting a white blood cell common antigen based on flow cytometry and application of the anti-CD45 antibody, the antibody comprises at least one of the following CDRs: a heavy chain variable region CDR: an amino acid sequence of SEQ ID NO: 1-3 or a conservatively modified form thereof; and a light chain variable region CDR: SEQ ID NO: 4-6 or an amino acid sequence in a conservative modification form thereof. The antibody or the antigen binding fragment thereof can efficiently and specifically recognize CD45, and can effectively detect the CD45.

According to the sorting control system of the flow cytometry sorting instrument, liquid drop images are shot through a camera, and liquid drop states are quantitatively evaluated and monitored on the basis of image content information. In order to determine the liquid drop breaking state, a liquid drop image stroboscopic shooting mode is designed to observe the liquid drop state, and the STM32H7 provides a synchronous control signal. When the droplet generation state is stable, the droplet breaking image is close to a stationary state. The stroboscope lamp is controlled to be turned on at different moments in a period, so that different states of liquid drop generation can be observed. The time difference between the detection time and the charging time is quantitatively determined through liquid drop delay control, and accurate delay control over sorting is achieved. And finally, a full-automatic algorithm is adopted to calibrate the liquid drop time delay, the liquid drop time delay is continuously tried to be changed, and when the shooting brightness of the sorted CCD camera under a certain liquid drop time delay is the highest, the time delay is considered to be the current liquid drop time delay, so that full-automatic calibration of the liquid drop time delay is realized.

The invention discloses a flow-type Raman sorting method and a high-luminous-flux flow-type Raman sorting device, and the method comprises the following steps: carrying out coincident focusing on Raman laser and near-infrared laser captured by a light trap to form a Raman excitation detection site, and arranging the Raman excitation detection site at the tip of the outer end of a tail end electrode in a chip dielectric focusing electrode; the photo-thermal oscillation near-infrared laser is focused on a tail end electrode in a dielectric focusing electrode of the chip and does not coincide with a Raman excitation detection site; through switching of light trap capture near-infrared laser and photo-thermal oscillation near-infrared laser, a single cell sample is captured and fixed at a Raman excitation detection site through light trap capture force generated by the light trap capture near-infrared laser or bubbles are generated through a photo-thermal effect generated by the photo-thermal oscillation near-infrared laser; the cell sample adhered to the tail end electrode in the dielectric focusing electrode is oscillated and separated through the bubbles. Compared with the prior art, the device has the beneficial effects that the organic combination of optical trap capture and dielectric capture sample oscillation functions can be realized, the switching is convenient, flexible, rapid, simple and reliable in use, the instrument cost is greatly reduced, the application range of the instrument is effectively widened, and the device has very strong universality.

The invention provides an automatic sample distribution method for flow cytometry, which belongs to the technical field of flow cytometry and comprises the following steps: acquiring basic information of a sample; the method comprises the following steps: pre-analyzing a sample through a micro-fluidic chip to obtain sample density, sample viscosity and sample ion concentration; measuring the micro-channel diameter and the micro-channel length of the micro-fluidic chip by adopting a laser ranging system, and recording the characteristic parameters of the micro-channel material; a target distribution volume is set, a high-precision injection pump system is started for pre-injection, and an initial distribution rate is obtained through measurement of a flow sensor; calculating the initial distribution rate by using a fluid dynamic compensation equation set to obtain a corrected distribution rate; controlling the high-precision injection pump system to perform sample distribution according to the corrected distribution rate; performing real-time monitoring on a sample liquid level by using an ultrasonic sensor, and synchronously acquiring a real-time distribution rate; and sample shunting is performed through the micro-fluidic chip, so that multi-channel parallel distribution is realized.

The invention discloses a method for improving the synthesis level of L-tryptophan in escherichia coli based on ARTP mutagenesis high-throughput screening, and belongs to the technical field of bioengineering. According to the invention, the L-tryptophan biosensor pSensor-trp3 is constructed, and the L-tryptophan biosensor pSensor-trp3 is good in response under the condition that the exogenous addition concentration range of L-tryptophan is 0-300mg. L <-1 >. High-throughput screening is performed by using a flow cytometry, a dominant mutant strain YB-2 is obtained through iterative mutagenesis, and the shake flask L-tryptophan yield and the sugar-acid conversion rate of the dominant mutant strain YB-2 are 5.68 g.L <-1 > and 0.103 g.g <-1 > respectively and are increased by 11.4% and 10.7%. Through second-generation whole genome re-sequencing comparative analysis, it is speculated that the genes ynfB and waaO possibly have a promoting effect on synthesis of L-tryptophan. The L-tryptophan biosensor used in the invention also provides reference for high-throughput screening and transformation of other related L-tryptophan production strains.

The invention relates to the field of plant breeding, and particularly provides a method for fixing rice heterosis by using an OsZFPP gene. The method comprises the following steps: firstly, constructing an OsZFPP1 gene or OsZFPP2 gene expression cassette driven by an OsECA1 promoter; then, an expression box of rice with three target points of OsPAIR1, OsREC8 and OsOSD1 being knocked out of CRISPR / Cas9 is constructed; integrating the expression cassette into the same vector to transform hybrid rice; screening three-gene homozygous mutation and OsZFPP positive plants, and identifying diploid cloned offspring through flow cytometry and genome sequencing. According to the method, the rice OsZFPP gene is combined with MiMe to provide a novel method for fixing the heterosis of the rice through apomixis, the heterosis of the rice can be successfully fixed through the method, an apomixis system with the high maturing rate can be obtained, and a novel solution is provided for fixing the heterosis of the rice through the apomixis.

The invention discloses a high-throughput antigen-specific T cell detection method based on mass spectrum flow cytometry. The invention belongs to the technical field of biology, and particularly relates to a high-throughput antigen-specific T cell detection method based on mass spectrum flow cytometry. According to the present invention, the activation induction marker (AIM) technology and the intracellular cytokine staining (ICS) are integrated, and the CyTOF platform is combined to achieve gt; 20 parameters are synchronously detected, and multiple activation-induction marker double positive markers, multifunctional cytokine spectrums (IFN-gamma, TNF-alpha, GZMB, IL-2 and the like) and immune memory phenotypes (CD45RA / CCR7) are covered. The standardized detection scheme provides a high-dimensional and quantifiable biomarker combination for vaccine protection efficacy evaluation, and has important clinical application value.

The invention discloses an auxiliary diagnosis system for renal transplantation rejection and typing thereof based on a peripheral blood immune cell map. Comprising a peripheral blood single cell acquisition unit, a mass spectrum flow cytometry unit, an original data preprocessing unit, a data conversion and normalization processing unit, a peripheral blood immune cell grouping acquisition unit, a peripheral blood immune cell map construction unit, a feature vector acquisition unit, a rejection prediction unit and a rejection typing prediction unit. According to the invention, a kidney transplantation two-stage diagnosis architecture is created for the first time; rejection detection and typing identification are realized through classification of an independently trained random forest binary classifier, and different TCMR / ABMR immune mechanisms are accurately matched; the traditional biopsy limitation is broken through, an auxiliary diagnosis scheme with high precision, low damage, moderate cost and typing identification capability is provided for the kidney transplantation rejection, and the method has important significance on timely discovery of the rejection of a kidney transplantation patient and improvement of prognosis.

The utility model relates to the technical field of cytometers, and discloses an automatic sampling mechanism of a flow cytometer, which comprises a bottom plate, a translation plate capable of horizontally moving is connected onto the bottom plate through a translation component, and a bearing component capable of rotating around the axis of the bearing component is connected onto the translation plate through a rotating component. A material loading part for clamping a test tube is mounted on the bearing part, and a uniform mixing assembly for uniformly mixing a sample in the test tube is also arranged on the bottom plate; the rotating assembly is mounted on the translation plate and can ensure that the bearing part can translate along a specified direction and can rotate around the axis of the bearing part, so that the material loading part can be automatically conveyed to the lower part of a corresponding sampling needle after clamping a test tube, and the interior of the test tube is mixed through the uniform mixing assembly; the operation precision and efficiency of the mechanism are both higher than those of manual operation, the labor intensity can be reduced, the tedious manual operation is liberated, the sample detection efficiency of flow cytometry is improved, and rapid sample loading and detection of the flow cytometry on samples are realized.

The invention discloses an anti-CD3D antibody and application thereof.The antibody comprises a heavy chain variable region and a light chain variable region, the amino acid sequence of CDR-H1 of the heavy chain variable region is shown as SEQ ID NO: 4, the amino acid sequence of CDR-H2 of the heavy chain variable region is shown as SEQ ID NO: 5, and the amino acid sequence of CDR-H3 of the heavy chain variable region is GDL; the amino acid sequence of the CDR-L1 of the light chain variable region is as shown in SEQ ID NO: 6, the amino acid sequence of the CDR-L2 of the light chain variable region is as shown in SEQ ID NO: 7, and the amino acid sequence of the CDR-L3 of the light chain variable region is as shown in SEQ ID NO: 8. The anti-CD3D antibody is a rabbit monoclonal antibody, the obtained antibody is high in titer, clear in specific positioning in immunohistochemical staining and free of non-specific background staining, the antibody is wide in application, and the antibody can be used in pathological immunohistochemical staining experiments and can also be used in Western blot and flow cytometry experiments.

The invention discloses an anti-soluble transferrin receptor monoclonal antibody as well as a preparation method and application thereof, and belongs to the technical field of antibody detection. According to the antibody, an L101-F760 amino acid sequence of human TfR protein is used as an immunogen, specific screening of B cells, reverse RNA (Ribonucleic Acid) into cDNA (Complementary Deoxyribose Nucleic Acid) through RT-PCR (Reverse Transcription-Polymerase Chain Reaction), then homologous recombination is performed to construct heavy and light chain vectors, and finally multiple rounds of screening are performed to obtain the purified sTfR monoclonal antibody, so that the sTfR protein can be specifically recognized, and the antibody has the advantages of high specificity and high sensitivity and can be widely applied to the field of immunotherapy. The natural sTfR protein in human serum can be recognized. The kit is applied to the detection and screening fields of immunohistochemistry, immunofluorescence chemistry, western blot, co-immunoprecipitation, indirect ELISA, flow cytometry and the like, especially has low background and no interference of non-specific binding impure protein during immunohistochemistry detection, can obtain more accurate detection and evaluation results, avoids interference of false positive and false negative, and has high detection accuracy. The method has great application value in the aspects of basic scientific research and clinical pathology detection.

The invention provides a multi-focus excitation confocal fluorescence microscopic imaging flow cytometry screening method. The method is characterized in that a laser beam generates three laser beams through a spatial light modulator, and the three laser beams are coherently superposed in a focal plane of a microscope objective to generate a two-dimensional optical lattice light field. And after the other laser beam is shaped by the cylindrical lens, linear laser optical tweezers are realized through the microscope objective, cells to be detected in an imaging detection area of the micro-flow chip are captured, and accurate active optical control on the rotation angle of the cells to be detected is realized. In a cell rotation process, a two-dimensional optical lattice light field rapidly scans in a focal plane at different angles, and a fluorophore in the cell is excited to generate a fluorescence signal. The confocal pinhole array installed at the position conjugate with the focal plane eliminates background fluorescence, a high-spatial-resolution fluorescence image of a three-dimensional structure in the cell is obtained, and cell screening is achieved. The method provided by the invention has the characteristics of high detection rate, high accuracy and the like, and has wide application prospects in the research fields of biology, medicine and life science.

SNAIL provides cost-efficient detection of specific nucleic acids in single cells, and may be combined with flow cytometry to simultaneously analyze large numbers of cells for a plurality of nucleic acids, e.g. at least one, to up to 5, up to 10, up to 15, up to 20 or more transcripts can be simultaneously analyzed, at a rate of up to about 50, 100, 250, 500 or more cells / second. The methods require only two primers for amplification, and may further include a detection primer.

The invention relates to a stroke prognosis prediction model training method based on quantum computing and artificial intelligence, and relates to the technical field of prognosis effect prediction. According to the method, leukocyte community parameters are introduced into an acute ischemic stroke prognosis model for the first time, morphological function characteristics of leukocytes are detected through a flow cytometry, and compared with traditional inflammation indexes, the leukocyte prognosis model has higher sensitivity and objectivity and becomes an important index for predicting neurological function impairment; the indexes closely related to cerebral apoplexy prognosis are screened out through quantum calculation for the first time, and a new thought is provided for high-dimensional data feature screening; meanwhile, by comparing 14 traditional machine learning, integrated learning and deep learning models, it is judged that the LightGBM model is an acute ischemic stroke prognosis prediction model, and compared with the prior art, the LightGBM model is remarkably improved.

The invention belongs to the field of intelligent equipment, and particularly relates to a system for automatically distinguishing leukemia types based on flow cytometry. The invention aims to solve the problem of low leukemia identification accuracy in the prior art. The invention provides a system for automatically distinguishing leukemia types based on flow cytometry. The system comprises a data acquisition module, a feature analysis module and a prediction and diagnosis module, the data acquisition module is used for processing bone marrow puncture fluid of a to-be-detected person to obtain a cell event data set of the to-be-detected person; the feature analysis module is used for processing a cell event data set of a to-be-detected person to obtain extraction features; the prediction and diagnosis module is used for processing extracted feature prediction to obtain a diagnosis conclusion; according to the system, through a standardized data preprocessing process, the analysis duration can be compressed to be within 20 minutes, the diagnosis consistency is improved to 98% or above, and the detection sensitivity reaches the 0.001% level.

The invention provides an antibody screening method for flow cytometry, and belongs to the technical field of flow cytometry. Antibody purity analysis and concentration determination are carried out through high performance liquid chromatography; the binding characteristic of the antibody and the target cell is researched by adopting an equal-ratio dilution method; analyzing the combined signal by using an improved Hilbert-Huang decomposition algorithm; evaluating the combination stability through covariance matrix decomposition; collecting cell event data by using a flow cytometer; constructing a fluorescence intensity contribution matrix and performing signal optimization; calculating a signal stability contribution value to determine an effective detection area; a 0-1 knapsack optimization method is adopted to carry out multiple detection combination optimization; finally, repeatability verification is carried out through a bootstrap resampling method, the stable-performance antibody for flow detection is screened out, and the technical problems that in the prior art, the signal stability of antibody screening for flow detection is poor, and specificity is insufficient are solved.

The invention discloses a group of blocking monoclonal antibodies of an SH1 homologous cluster fully human thyrotropin receptor (TSHR) (a TSH receptor, TSHR) as well as a preparation method and application of the blocking monoclonal antibodies of the SH1 homologous cluster fully human thyrotropin receptor (TSHR). The method comprises the following steps: sorting plasma cells and memory single B cells for specifically recognizing TSHR in peripheral blood of a patient with high TSH receptor blocking antibody (TBAb) titer by using flow cytometry, cloning antibody light and heavy chains in vitro and performing recombinant expression, and performing antibody property screening verification by using hTSHR-CHO cells. The blocking monoclonal antibody specifically targeting the human TSHR is obtained. The fully human TSH receptor blocking monoclonal antibody disclosed by the invention can be specifically combined with TSHR, and can effectively block signal transduction after the TSH is combined with the receptor; the synthesis and secretion of thyroid hormone are inhibited; expression and fibrosis of effector cell orbital fibroblasts TSHR for inhibiting thyroid-related eye diseases are remarkably relieved, and wide application prospects are achieved in treatment of Graves' diseases and diseases such as thyroid eye diseases caused by hyperthyroidism.

According to the exosome multi-target detection method based on the nano-frame multi-satellite mode, a high-stability tetrahedron is used as a core skeleton, multiple high-protein-affinity nucleic acid aptamer tongs are assembled and matched according to needs to construct a high-reproducibility biomarker recognition element, a molecular form with multiple satellites wrapping exosome is constructed, and the exosome multi-target detection method based on the nano-frame multi-satellite mode is obtained. According to the method, a'free satellite 'element is formed, and flow cytometry is utilized to detect the particle size and output signals with ultralow background interference, so that multiple detection requirements of different tumors on different expression levels of different markers in different development processes can be met, and an identification-separation-detection integrated high-specificity exosome detection strategy is realized. Clinical examination and diagnosis requirements serve as fundamental guidance, detection potential is excavated on an existing basic flow platform, instrument detection particle size limitation is turned into background killer, a new instrument detection thought is widened, detection performance is improved, exosome encrypted information decoding is accelerated, and the transformation progress of clinical disease prevention and treatment and tumor diagnosis and treatment is promoted and guided.

The invention discloses a TMPRSS2 protein inhibitor efficacy prediction calculation method. A scoring function is constructed through a logP value, a virtual docking score and living cell inhibitor effect data to predict a drug inhibition effect. The method specifically comprises the following steps: constructing a TMPRSS2 overexpression cell model to screen drugs; the cell viability of various potential inhibitors is detected, and high-toxicity inhibitors are eliminated through a CCK-8 experiment; using flow cytometry to detect the cell average fluorescence intensity through a TMPRSS2 fluorescent probe to evaluate the enzyme activity influence of the inhibitor, and calculating the IC30 value; measuring the logP value of an effective inhibitor, and predicting the binding capacity of the effective inhibitor and TMPRSS2 by using Schrodinger software to obtain a virtual docking score; and analyzing the relationship among the IC30 value, the logP value and the virtual docking score, and constructing a scoring function to predict the drug effect. According to the method, the drug effect prediction can be simply, conveniently and quickly carried out, the drug screening consumption is reduced, and the drug screening period is shortened.

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 invention discloses a melatonin detection method which comprises the following steps: S1, coupling an anti-melatonin antibody with a PE fluorescent label with carboxyl fluorescent magnetic beads to construct antibody coupled magnetic beads MBs (at) Ab; s2, preparing a melatonin analogue FITC-MT by using the 5-MT and the FITC-NHS (N-hydroxysuccinimide); s3, preparing MBs (at) Ab and FITC-MT into a reaction system, reacting with melatonin standard substances with different concentrations or a to-be-detected sample, detecting by using a flow cytometer, processing a detection result of the flow cytometer by using a machine learning algorithm, and calculating the melatonin concentration in the to-be-detected sample. MT in a to-be-detected sample can compete for FITC-MT combined with MBs (at) Ab in a short time to form a stable reaction system, the multi-dimensional features of MBs (at) Ab are subjected to flow detection to directly remove impurities, agglomeration and the like in MBs (at) Ab, different fluorescence features of an optimal magnetic bead group in the system are searched and analyzed in combination with a machine learning algorithm, and the content of MT can be accurately detected; the reaction system constructed by the invention is simple to operate and extremely low in cost, and meanwhile, the result accuracy can be ensured.

The invention discloses a group of blocking monoclonal antibodies of a CHK36 homologous cluster fully human TSH receptor (TSHR) as well as a preparation method and application of the blocking monoclonal antibodies. The method comprises the following steps: sorting plasma cells and memory single B cells for specifically recognizing TSHR in peripheral blood of a patient with high TSH-blocking antibody (TBAb) titer by using flow cytometry, cloning antibody light and heavy chains in vitro and performing recombinant expression, and performing antibody property screening verification by using hTSHR-CHO cells, so as to obtain the TSHR-TBAb-TSHR-TBAb. The blocking monoclonal antibody specifically targeting the human TSHR is obtained. The fully human TSH receptor blocking monoclonal antibody disclosed by the invention can be specifically combined with TSHR, and can effectively block signal transduction after the TSH is combined with the receptor; the synthesis and secretion of thyroid hormone are inhibited; according to the present invention, with the application of the compound, the expression and the fibrosis of the effector cell orbit fibroblast TSHR inhibiting the thyroid-related eye disease can be significantly reduced, and the wide application prospect is provided in the treatment of Graves' disease (Graves' disease, GD) and diseases caused by hyperthyroidism, such as thyroid eye disease (thyroid eye disease, TED), and the like.

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 invention relates to the technical field of insulin resistance, in particular to an insulin resistance method for promoting a signal channel based on CTRP9. The method comprises the following steps: constructing a human hepatocyte line plasmid; carrying out cell culture and transfection; detecting the influence of CTRP9 on LO human hepatocyte glucose uptake, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; detecting the influence of CTRP9 on glucose uptake of an LO human hepatocyte insulin resistance model, PGC-1a gene expression and the phosphorylation level of insulin signal channel related protein; and carrying out SirT1 / PGC-1a protein expression detection. According to the insulin resistance method for promoting the signal channel based on the CTRP9, a human hepatocyte line is taken as an in-vitro research object, an in-vivo research environment is simulated, a corresponding insulin resistance model is established, and the insulin resistance can be obtained by using technologies such as flow cytometry analysis, RT-PCR (Reverse Transcription-Polymerase Chain Reaction), western blotting and signal channel inhibition. And the treatment effect of CTRP9 on hepatic cell insulin resistance and the specific action mechanism are researched.

The invention provides an application of a reagent for detecting an effector CD4 + T cell in peripheral blood of a subject in preparation of a product for diagnosing membranous nephropathy, lupus nephritis or ANCA-related vasculitis. The invention also provides an application of the effector CD4 + T cell in peripheral blood in preparation of a diagnostic model for diagnosing membranous nephropathy, lupus nephritis or ANCA-related vasculitis. The marker is detected through flow cytometry, and the result shows that the Ejector CD4 + T cell is remarkably increased in a patient with membranous nephropathy, can effectively distinguish membranous nephropathy from healthy control (verification set AUC 0.8098), can assist in identifying membranous nephropathy, lupus nephritis (verification set AUC 0.8686) and ANCA-related vasculitis (verification set AUC 0.8968), and the level of the Ejector CD4 + T cell is related to whether membranous nephropathy is difficult to treat or not.