Cell staining reagent and preparation method and application thereof in cell staining

Abstract

The invention provides a cell staining reagent, comprising B.S liquid, staining liquid, rinsing liquid, and EA50 staining liquid; wherein per 400ml of B.S liquid consists of 320.0ml of methanol, 60.0ml of formol and 20.0ml of acetic acid glacial; per 210ml of staining liquid comprises 100.00mg of thiohydrazine, 87.0mg of tert-butanol liquid, 5.2ml of hydrochloric acid with the concentration of 5mol / L, 1800mg of sodium bisulfite, and the balance of distilled water; per 200ml of rising liquid comprises 1.0g of sodium metabisulfite, 2.0ml of hydrochloric acid with the concentration of 5mol / L, and 198.0ml of distilled water; per 600ml of EA50 staining liquid comprises 15ml of purified water, 0.3g of bright green, 2g of water soluble eosin Y, 450ml of ethanol with volume percent of 95 percent, 125ml of pure methanol, 10ml of glacial acetic acid, and 1g of phosphotungstic acid. The invention further provides the preparation method and the application of the cell staining reagent in cell staining.

Description

technical field [0001] The invention relates to a cell staining reagent capable of simultaneously staining cell nuclei and cytoplasm with different colors, its preparation method and application in cell staining. technical background [0002] For nearly a century, with the development of medicine and the deepening of pathological research, the qualitative and quantitative detection of cells has become a common method in medicine. [0003] Cell morphology analysis is a method of qualitative detection of cells, by observing the morphological characteristics of the exfoliated cells under a microscope to qualitatively analyze the cells, so as to provide reference for further pathological analysis. In order to observe the cells, it is usually necessary to stain the whole cell first, so that the cell membrane, cytoplasm and nucleus can be easily observed under a microscope. Staining method, Shaw staining method, Maggie staining and so on. There are only a few descriptive cellula...

AI Technical Summary

Problems solved by technology

However, this method is limited by the quality of DNA fixation, the depth of staining, and the quality of the measurement system (scanning microscope, digital camera, application software, etc.), and this method does not use cell morphology information, which is It has a great effect on the diagnosis and prediction of many cases

[0006] Therefore, in general, these two analysis methods (qualitative and quantitative) have the following problems: 1. The method of qualitative analysis of cell morphology is only based on the morphological characteristics of cells, and relies on the experience and skills of cytologists, and the misdiagnosis rate is very high

2. DNA quantitative analysis methods are based on DNA-specific staining methods, which depend on the characteristics of the nucleus and cannot provide any information about cell morphology

[0007] So far, there is no method that can simultaneously perform qualitative analysis of morphological characteristics of cells and quantitative analysis of DNA. However, in the process of diagnosis of many diseases, in order to improve the accuracy, it is necessary to perform both qualitative and quantitative detection at the same time, so it is necessary to Slicing and staining twice is not only very troublesome, but also increases the cost burden of patients. Moreover, the results of qualitative and quantitative testing cannot be one-to-one. Cells, when you want to obtain the DNA quantitative analysis data of the cells for control confirmation, it is not possible, because the corresponding cells cannot be found in the section for DNA quantitative analysis for analysis

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