30 results about "Antibody staining" patented technology

The invention belongs to the technical field of kidney histopathology and immunological evaluation, and particularly relates to a kidney pathology detection kit, a detection method and application, the detection kit comprises 37 rare metal element labeled monoclonal antibodies combined with antigens of in-situ tissue cells, and the detection method comprises the following steps: (1) collecting a sample to be detected, preparing a pre-dyed paraffin section; (2) mixing 37 monoclonal antibodies labeled by rare metal elements, and carrying out antibody staining on various antigen targets on the surfaces of cells in the in-situ tissue; (3) carrying out nucleus DNA staining on the stained tissue sample; and (4) detecting the stained tissue sample through an imaging mass spectrum flow system, and collecting original information data through laser spot firing. The method is an analysis method for fine grouping identification and relative quantification of structural cells, immune cells and pathological injury cells in the clinical nephropathy sample in-situ tissue. And auxiliary diagnosis information is provided for diagnosis, grading, treatment and prognosis detection of various clinical nephropathy.

The invention relates to a method for detecting a B cell subset in a lymphatic tissue, a kit comprising a quantum dot-antibody CD19 conjugate, a quantum dot-antibody CD138 conjugate and a quantum dot-antibody CD1d conjugate is used for dyeing a lymphatic tissue section, and different antibodies are marked by quantum dots with different colors, so that the B cell subset in the lymphatic tissue section can be detected, and the B cell subset in the lymphatic tissue section can be detected. Content and positions of plasma cells and regulating B cells are confirmed, so that biological behaviors of the plasma cells and regulating B cells can be determined conveniently. According to the method, the existing multi-color immunofluorescence technology is improved, a secondary antibody dyeing link is omitted by using a quantum dot carrying antibody method, a direct method is adopted for marking, the advantages of quantum dots are exerted, the experiment time is saved, and the efficiency is improved.

The invention relates to a method for detecting Breg cells and Tfh cells in a chest adenoma tissue microenvironment, which comprises the following steps of: coupling quantum dots with a specific antibody, dyeing a chest adenoma tissue slice sample, and specifically connecting the specific antibody with antigens on the Breg cells and / or Tfh cells in the slice sample cells, so as to quantify or position the Breg cells and / or Tfh cells by the quantum dots. According to the method disclosed by the invention, the existing multicolor immunofluorescence technology is improved, and a secondary antibody dyeing link is omitted by using a method of carrying the antibody by the quantum dots, so that the efficiency is improved; the quantum dots are used for replacing fluorescent dye, direct marking of tissue slices can be achieved, local positioning of Breg cells and Tfh cells in a thymoma tumor microenvironment is achieved, and biological behaviors of the Breg cells and the Tfh cells can be determined conveniently; the immune condition can be evaluated by detecting the ratio condition of Breg / Tfh cells in the breast adenoma tumor microenvironment of a patient in vitro.

A method for evaluating a biological material for unassociated virus-size particles having a particular epitope uses a fluorescent antibody stain specific for binding with the epitope and a fluid sample with the virus-size particles and fluorescent antibody stain is subjected to flow cytometry with identification of fluorescent emission detection events indicative of passage through a flow cell of a flow cytometer of unassociated labeled particles of virus size including such a virus-size particle and fluorescent antibody stain.

The invention relates to the technical field of medical examination, in particular to a method for rapidly detecting a neutrophil extracellular trapping net. The method comprises the following steps: (1) taking a human whole blood sample, adding a hemolytic agent, then adding a mixed fluorescent antibody staining solution, shaking and uniformly mixing, and standing and incubating at room temperature; (2) centrifuging after incubation in the step (1), absorbing and discarding a supernatant, washing with a washing solution, centrifuging, absorbing and discarding the supernatant; (3) adding a washing solution to resuspend the cells; and (4) detecting by adopting a flow cytometer. The detection method is good in stability, high in repeatability and high in fluorescence intensity, and can be used for auxiliary diagnosis of diseases.

The invention relates to a kit for detecting plasma cells and related antibody levels in a thymus microenvironment and application of the kit. The kit comprises a quantum dot labeled antibody CD138, a quantum dot labeled antibody CD21, a quantum dot labeled AchR, Musk and / or LRP4 receptor, an antigen repair solution and a DNA fluorescent dye solution, wherein the quantum dot labeled antibody CD138, the quantum dot labeled antibody CD21, the quantum dot labeled AchR, the quantum dot labeled Musk and / or the quantum dot labeled LRP4 receptor, the antigen repair solution and the DNA fluorescent dye solution; through the kit, one or more of plasma cells, follicular DC and related antibodies in a thymus (tumor) sample can be detected, so that the immune level of a tissue slice sample is obtained. A method of carrying the antibody by using the quantum dots is adopted, so that a secondary antibody dyeing link in the existing multicolor immunofluorescence technology is omitted, and the efficiency is improved; the quantum dots are used for replacing fluorescent dye, direct marking of tissue slices can be achieved, local positioning of plasma cells and follicular DCs in the thymoma tumor microenvironment is achieved, and both quantity detection and position detection can be achieved.

The present invention relates to the field of medical testing technology, and in particular to a method for rapidly detecting neutrophil extracellular traps. The method comprises the following steps: (1) taking a human whole blood sample, adding a hemolytic agent, and then adding a mixed fluorescent antibody staining solution, shaking and mixing, and incubating at room temperature; (2) centrifuging after incubation in step (1), discarding the supernatant, washing with a washing solution, and then centrifuging, discarding the supernatant; (3) adding a washing solution to resuspend the cells; (4) using a flow cytometer for detection. The detection method has good stability, strong repeatability, and high fluorescence intensity, and can be used for auxiliary diagnosis of diseases.

The application discloses a method for separating rat intestinal nerve cells, takes the intestinal muscle layer of a rat intestine and digests it into single cells, and then purifies the rat intestinal nerve cells by performing flow antibody staining and flow cytometry sorting on cell surface molecules such as CD9 and CD45, and comprises the following general steps: firstly, single cell preparation, secondly, single cell staining, and finally, flow cytometry separation. The application has a short time consumption and can realize real-time separation; the cell surface molecule markers for the rat intestinal nerve cells are determined to be CD45 negative and CD9 positive, and the rat intestinal nerve cells can be rapidly separated in real time for scientific research and clinical use.

The application relates to the technical field of immunodetection, in particular to a non-crystalline MOF-mediated cascade antibody directional probe and a preparation method and application thereof. The application specifically comprises the following steps: (1) mixing a second antibody solution with a coomassie brilliant blue solution to obtain a second antibody staining solution; (2) mixing a Zn(OAc)2.2H2O solution, an HmIM solution and the second antibody staining solution to obtain aR-ZIF-Ab2 nanocomposite material; and (3) mixing the aR-ZIF-Ab2 nanocomposite material with a first antibody and incubating to obtain aR-ZIF-Ab2-mAb1 cascade antibody directional probe. The CAbD strategy based on aMOF can be used as a promising antibody directional and probe construction means in immunochromatographic analysis, and provides a reference innovation direction for the design and development of an immunobiosensor.

Embodiments of the present invention encompass systems and methods for determining detection limits for various antibody-dye conjugates for flow cytometry. Exemplary techniques involve a linear superpositioning approach of spillover-induced enlargements of normally distributed measurement errors.

A method for evaluating a biological material for unassociated virus-size particles having a particular epitope uses a fluorescent antibody stain specific for binding with the epitope and a fluid sample with the virus-size particles and fluorescent antibody stain is subjected to flow cytometry with identification of fluorescent emission detection events indicative of passage through a flow cell of a flow cytometer of unassociated labeled particles of virus size including such a virus-size particle and fluorescent antibody stain.

The invention discloses an analysis method for detecting the H3K27me3 level of each immune cell population in human whole blood by using flow cytometry, which comprises the following steps of: by a pretreatment mode of splitting red blood cells in a human whole blood sample by using a one-step method and fixing white blood cells, dyeing the split and fixed white blood cells by using fluorescently labeled antibodies (BV785-CD3, BV650-CD19, BV605-CD14, AF488-HLA-DR, AF647-H3K27 and Pacilic Blue-H3); and carrying out gate analysis on the sample after antibody staining by using a flow cytometer, and calculating the H3K27me3 / H3 level in each immune cell population, namely T cells (CD3 + CD19-), B-cell (CD3-CD19 +), mononuclear cells (CD14 + HLADR +) and granulocytes (CD14-HLADR-) through a formula. The whole blood sample can be preserved for 5 days under the condition of-60 to-90 DEG C after being subjected to one-step cracking and curing treatment. The method has good accuracy, reliability and stability.

This invention discloses a three-dimensional imaging method for biological tissue samples based on CUBIC clearing antibody staining, comprising: placing mouse brain tissue in CUBIC-1 solution for degreasing and dehydration; subjecting the degreased and dehydrated mouse brain tissue to sequential cell membrane permeation and blocking treatments; incubating the blocked mouse brain tissue with diluted primary antibody GFAP and secondary antibody Alexa488; washing the mouse brain tissue after two antibody incubations; placing the washed mouse brain tissue in CUBIC-2 solution for matching, and then embedding the mouse brain tissue in resin; placing the resin-embedded sample in a buffer bath, and acquiring images using a TDI-fMOST camera. This invention achieves overall staining of large biological tissue samples using CUBIC clearing antibody staining.

The present invention provides a method for cell analysis, comprising: preparing a blood sample comprising nucleated cells having surface, cytoplasmic or nuclear antigens (markers); antibody staining the cell markers; fixing and permeabilising the cells; FISH probe hybridising to chromosomes in the cells; performing imaging flow cytometry on the cells; analysing data obtained from performing imaging flow cytometry; and diagnosing, prognosing or monitoring a medical condition based on the data analysis.

The present invention relates to a method for detecting B cell subsets in lymphoid tissue, wherein lymphoid tissue sections are stained by a kit including quantum dot-antibody CD19 conjugate, quantum dot-antibody CD138 conjugate and quantum dot-antibody CD1d conjugate, and different antibodies are marked by quantum dots of different colors to realize the confirmation of the content and position of B cells, plasma cells and regulatory B cells in tissue sections, so as to facilitate the clarification of their biological behaviors. The method of the present invention improves the existing multicolor immunofluorescence technology, uses a method of carrying antibodies with quantum dots, eliminates the secondary antibody staining link, adopts direct labeling, and gives full play to the advantages of quantum dots themselves, saves experimental time, and improves efficiency.

The present invention discloses a method for cell immunofluorescence staining capable of amplifying signals, belonging to the field of biotechnology. In the present invention, the aptamer HCR primer is anchored to the cell surface target, and an HCR reaction is carried out to extend a fluorescent HCR product chain, thereby realizing cell immunofluorescence staining. Compared with the traditional antibody staining technology, the fluorescence signal intensity of the present invention is increased by 1.5 times.

This invention discloses an analytical method for detecting the H3K27me3 level of various immune cell populations in human whole blood using flow cytometry. The method involves a one-step pretreatment process of lysing erythrocytes and simultaneously fixing leukocytes in a human whole blood sample. Fluorescently labeled antibodies (BV785-CD3, BV650-CD19, BV605-CD14, AF488-HLA-DR, AF647-H3K27, Pacific Blue-H3) are used to stain the lysed and fixed leukocytes. After antibody staining, the sample is subjected to flow cytometry for gating analysis. The H3K27me3 / H3 level in each immune cell population—T cells (CD3+CD19-), B cells (CD3-CD19+), monocytes (CD14+HLADR+), and granulocytes (CD14-HLADR-)—is calculated using a formula. The whole blood sample, after the one-step lysing and fixing process, can be stored at -60 to -90°C for 5 days. The method exhibits good accuracy, reliability, and stability.

The invention provides a method for removing background fluorescence of a tissue sample, which comprises the following steps: mixing the tissue sample with a metal hydride reducing agent, and illuminating the tissue sample under white light and a light source with the wavelength of 475-485 nm. According to the method, the background fluorescence of the tissue sample can be obviously removed, the background is cleaner after the treated tissue is subjected to antibody dyeing, the strength and expression quantity of an antibody labeling signal are not influenced, the signal-to-noise ratio of a specific fluorescence labeling signal is high, the removal effect is thorough, and the background fluorescence cannot be recovered. Meanwhile, the method disclosed by the invention is low in cost and simple and convenient to operate, and has a wide application prospect in technologies such as immunofluorescence analysis and the like.

The present invention discloses a method for preparing a cell smear, which comprises the steps of preparing a cell suspension, fixing cells, staining cells and drying cells, wherein the cell drying is to culture the cell smear at 50°C under aeration conditions for 40-60 minutes. The present invention also discloses the use of the cell smear in the preparation of a drug for preventing and treating a disease. The preparation method of the present invention can maintain or improve the cell recovery rate, retain the complete cell nucleus type and not affect the membrane protein antibody staining, and can also reduce the drying time and speed up the overall experimental progress.

The present invention relates to a method for detecting Breg cells and Tfh cells in a thymoma tissue microenvironment, wherein quantum dots are coupled with specific antibodies, and thymoma tissue slice samples are stained. After the specific antibodies are specifically connected to antigens on Breg cells and / or Tfh cells in the slice sample cells, quantum dots are used to quantify or locate Breg cells and / or Tfh cells. The method of the present invention improves the existing multicolor immunofluorescence technology, and uses a method of carrying antibodies with quantum dots, which eliminates the secondary antibody staining link and improves efficiency; by replacing fluorescent dyes with quantum dots, direct labeling of tissue slices can be achieved, and local positioning of Breg cells and Tfh cells in the thymoma tumor microenvironment is achieved, which is convenient for clarifying their biological behaviors; and the immune status can be evaluated by in vitro detecting the ratio of Breg / Tfh cells in the thymoma tumor microenvironment of the patient.

The invention discloses a rat intestinal nerve cell separation method, which comprises the following steps: taking a rat intestinal muscle layer, digesting the rat intestinal muscle layer into single cells, carrying out flow antibody staining and flow cytometry sorting on cell surface molecules such as CD9 and CD45, and purifying the rat intestinal nerve cells, and comprises the following steps: firstly, preparing the single cells, secondly, staining the single cells, and finally, separating the rat intestinal nerve cells. And finally, separating by flow cytometry. The method is short in time consumption, and real-time separation can be realized; the cell surface molecular tags aiming at the rat intestinal nerve cells are determined to be CD45 negative and CD9 positive, and the rat intestinal nerve cells can be rapidly separated in real time for scientific research and clinical application.

The invention relates to a use method and application of a mitochondrial membrane potential probe. The use method comprises the following steps: (a) separating human peripheral blood mononuclear cells by adopting a Ficoll method; (b) carrying out flow antibody staining; (c) centrifugally collecting the cells, cleaning with a PBS buffer solution, and adding a cell culture medium to resuspend the cells; (d) adding a fluorescent probe JC-1 into the culture medium, incubating in a 5% CO2 cell incubator at 37 DEG C for 15-30 minutes, centrifuging, precipitating cells, and discarding the supernatant; adding resuspended cells, centrifuging, precipitating the cells, and discarding the supernatant; and (e) resuspending the cells in a precooled PBS buffer solution, and measuring and analyzing the membrane potential in a flow cytometer. The progress of the liver disease can be analyzed according to the significant difference, the accuracy and specificity of diagnosis are significantly improved, and a reliable basis is provided for clinical diagnosis.

The present invention belongs to the technical field of biochemical detection, and specifically relates to a kit and application for multiple immunoassays. It can be used for multi-target detection of circulating tumor cells in different body fluids such as blood, or for projects involving multi-target detection in tissue sections, cell slides and other detection processes. It is particularly suitable for multi-target staining experiments of antibodies of the same species but different subtypes. By digesting the anti-mouse IgG antibodies obtained by goat immunization with papain, purifying the Fab fragments and labeling them with dyes, a GM Fab secondary antibody with subtype recognition ability is prepared, which is used for specific blocking operations after multiple rounds of primary antibody staining, significantly reducing cross-interference between different primary antibodies. After multiple staining verification in tumor tissues and circulating tumor cells after blood enrichment, the results show that the signal distribution of each target is clear, the positioning is accurate, and the background interference is low, which is suitable for tumor immune microenvironment analysis and immunotherapy evaluation.

Fluidic systems, devices and methods are provided for separating and sequestering particles from a fluid flow within traps in a fluidic chip is provided. The fluidic platform is particularly suited for parallel live zebrafish embryo studies providing automated zebrafish embryo trapping and flow through culture as well as whole mount zebrafish antibody staining functions. The zebrafish on a chip testing platform uses a chaotic hydrodynamic trapping process to trap and retain zebrafish embryos in a consistent body orientation (i.e., head pointed inward) without any external adjustments. The system and apparatus can also be adapted to be a multifunctional concentration gradient generator (CGG) that can be used to automatically immobilize dechorionated zebrafish embryos and generate chemical gradients for acute fish embryo toxicity (FET) tests.

The invention belongs to the technical field of biology, and particularly relates to a dyeing method and application thereof. The staining method comprises the following steps: placing a staining reagent or one or two of the staining reagent and an intermediate on a biological sample in the form of a liquid film; compared with a traditional dyeing method and a microfluidic dyeing method, the method can greatly improve the dyeing efficiency, shorten the dyeing time and save the dyeing reagent on the premise of keeping the dyeing effect. The method has wide potential application value, including fluorescent antibody dyeing, nucleotide coupled antibody dyeing, in-situ hybridization, aptamer dyeing and the like. The method can be used for animal and plant cell distribution, construction of tumor and disease maps, and screening and discovery of digital pathology and biomarkers.

The invention relates to a device for removing background fluorescence of a tissue sample, the device comprises a first shell, a light source module, a sample module, a cooling module and a control module, the light source module and the cooling module are both mounted in an inner cavity of the first shell; the sample module can mix a tissue sample with a reducing agent and then fix the mixture in an inner cavity of the first shell, and the reducing agent comprises a metal hydride reducing agent; the light source module can emit a light source for irradiating the sample module, and the light source and the reducing agent act on the tissue sample to remove background fluorescence of the tissue sample; the cooling module is used for cooling the inner cavity of the first shell; the control module is connected with the cooling module and the light source module. According to the scheme, background fluorescence of a tissue sample can be obviously removed, the background is cleaner after the treated tissue is subjected to antibody staining, and the signal fidelity is improved; the removal effect is thorough, background fluorescence cannot be recovered, and the stability is high; the method basically has no influence on tissue morphology, is basically suitable for all tissues of animals and plants, is not limited by types, and is high in universality.

The present invention relates to a kit for detecting plasma cells and related antibody levels in a thymic microenvironment and its application. The kit includes quantum dot-labeled antibody CD138, quantum dot-labeled antibody CD21, quantum dot-labeled AchR, Musk and / or LRP4 receptors, antigen repair solution and DNA fluorescent dye; the kit can detect one or more of plasma cells, follicular DC and related antibodies in a thymic (tumor) sample, so as to obtain the immune level of a tissue section sample. The method of using quantum dots to carry antibodies eliminates the secondary antibody staining link in the existing multi-color immunofluorescence technology and improves efficiency; by replacing fluorescent dyes with quantum dots, direct labeling of tissue sections can be achieved, and local positioning of plasma cells and follicular DC in the thymoma tumor microenvironment can be achieved, which can achieve both quantity detection and position detection.

The invention belongs to the technical field of biochemical detection, and particularly relates to a kit for multiple immunodetection and application thereof, and the kit can be used for multi-target detection of circulating tumor cells in different body fluids such as blood, or items related to multi-target detection in tissue slices, cell slide and other detection processes. And the method is particularly suitable for multi-target staining experiments of different subtype antibodies of the same species. An anti-mouse IgG antibody obtained through goat immunization is subjected to papain digestion, Fab fragment purification and dye labeling, a GM Fab secondary antibody with subtype recognition capacity is prepared and used for specific closing operation after multiple rounds of primary antibody staining, and cross interference among different primary antibodies is remarkably reduced. Multiple staining verification is carried out on circulating tumor cells in tumor tissues and after blood enrichment, and the result shows that each target signal is clear in distribution, accurate in positioning and low in background interference, and is suitable for tumor immune microenvironment analysis and immunotherapy evaluation.

The invention relates to a method for detecting immune reconstruction of AIDS patients through full-spectrum flow cytometry, belongs to the technical field of immune cell detection, and is used for detecting various immune cells of peripheral blood mononuclear cells of healthy individuals, immune responders and immune non-responders on the basis of full-spectrum flow cytometry. The method comprises the following steps: carrying out antibody staining labeled by 28 fluorescein in a cell sample, distinguishing 25 immune cell subgroups including NK cells, B cells and T cells, detecting immune function markers of the immune cell subgroups, and analyzing the quantity proportion and functional states of the immune cell subgroups of HIV patients with different immune reconstruction degrees.