Micro-immunofluorescence detection method without cell loss

Abstract

The invention discloses a micro-immunofluorescence detection method without cell loss, which includes the following steps: sterilizing 0.22um Spin-X Centrifuge Tube; transferring cells to the sterilized 0.22um Spin-X Centrifuge Tube to directly perform subsequent steps. Or perform subsequent steps after culture; subsequent steps include: PBS buffer washing, 4% paraformaldehyde fixation, methanol treatment on ice, 0.3M glycine blocking solution treatment, 4% BSA blocking solution treatment, fluorescent primary antibody incubation at room temperature in the dark, Stain with DAPI staining solution, place the filter membrane on a glass slide, seal it, and observe it under a laser scanning confocal microscope. This method makes up for the shortcomings of immunofluorescence technology that is difficult to accurately screen specific cells, and provides a simple, accurate and visual technology for screening specific cells in mixed cell populations.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a micro-immunofluorescence detection technique without cell loss. Background technique [0002] Immunofluorescence technique (immunofluorescence technique) is a method of combining immunological methods (antigen-antibody specific binding) with fluorescent labeling technology to study the distribution of specific protein antigens in cells. Its principle: first mark known antigens or antibodies with fluorescein to make fluorescent markers, and then use this fluorescent antibody (or antigen) as a molecular probe to check the corresponding antigen (or antibody) in cells or tissues. The antigen-antibody complexes formed in cells or tissues contain fluorescein. Use a fluorescence microscope to observe the specimen. The fluorescein emits bright fluorescence (yellow-green or orange-red) under the irradiation of excitation light, and the cells or tissues where the fluorescence is ...

AI Technical Summary

Problems solved by technology

At the same time, in the process of cell labeling and fluorescence, it has to undergo multiple washings, which cannot avoid the loss of cells during the test, and it is difficult to identify a small amount of special cells in the sample, thus limiting the application of immunofluorescence technology in cell screening.

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