Liquid phase staining identification method used for cells and needing extremely small cell quantity

Abstract

The invention provides a liquid phase staining identification method used for cells and needing extremely small cell quantity. The method comprises the following steps: (1) adding a cell immobilizingsolution into cells in a non-dry state for cell immobilization; (2) after cleaning the cells, adding a cell staining solution for cell staining, wherein the cell staining solution comprises a fluorescent antibody staining solution; (3) dyeing the cells by utilizing bright-field light dyeing liquid; and (4) after cleaning the cells, suspending the cells, and observing the cells under fluorescence and bright-field light via a microscope so as to simultaneously obtaining various fluorescence and bright-field light images. According to the invention, on the premise that cells are not dried or sealed, staining and imaging via dual light sources of fluorescence antibody and bright field optical cell morphology can be carried out, and high-quality fluorescence markers and bright-field optical morphological images of the cells can be obtained; and the method disclosed by the invention is simple and feasible in process, low in cost and beneficial for cell classification and subsequent molecular-level analysis, and more comprehensive cell marker information can be obtained.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a cell liquid phase staining identification method with a very small amount of cells. Background technique [0002] In recent years, with its advantages in tumor detection and analysis, liquid biopsy technology has solved many limitations of surgical pathology and tissue puncture pathology, improved the sensitivity of early detection and advanced the time of early detection by 3-5 years. The fields of pharmaceuticals and biotechnology play an increasingly important role. As one of the most important markers in liquid biopsy, the detection of circulating tumor cells plays an important role in early detection, counting and gene analysis because of its advantages as a liquid biopsy marker that can best represent the information of the primary tumor and diffuse foci. Important and essential functions such as diagnosis, disease tracking, and recurrence detection. However, ci...

AI Technical Summary

Problems solved by technology

The problem is that with this method, it is impossible to ensure the integrity of a single cell collected by this method. Then, in the subsequent genetic analysis, it is due to the gene deletion of the original tumor cells, or the lack of cell integrity during the collection of cells. If there are many cells, it is easy to lose the main gene components when performing subsequent DNA / RNA extraction, which will cause the results to lose accuracy and cause a lot of confusion in clinical applications.

Most of the existing staining methods on the market rely on staining on glass slides after drying, which is difficult to implement in liquid phase, and there is no method that can simultaneously stain single-cell fluorescence and bright-field light at the same time. In the case of fluorescent staining, there is still the difficulty of insufficient signal, so there is no suitable technology and product on the market

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