8 results about "Antibody staining" patented technology

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.

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.

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 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 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 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.