DNA quantitative staining method for anti-slide-off cells

A staining method and cell technology, which is applied in the field of DNA quantitative staining of anti-dropping cells, can solve the problems of unresolved influence of anti-dropping glass slides, reduced accuracy, and cures the symptoms rather than the root causes, and achieves high commercial value and guarantees. Completeness and wide applicability

Active Publication Date: 2020-11-27
湖南品胜生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, due to the cumbersome staining process of the traditional method, it is easily affected by the type of tissue fixative, acid hydrolysis time and temperature, and only by enhancing the adhesion to prevent the slide from falling off is not a cure for the symptoms. occur
The influence of cold, heat, acid, alkali and alcohol on the anti-shedding slides in the methodology has not been fundamentally resolved
Due to the low specificity of basic Schiff, the background is prone to co-staining, which affects the judgment of the results and reduces the accuracy

Method used

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  • DNA quantitative staining method for anti-slide-off cells
  • DNA quantitative staining method for anti-slide-off cells
  • DNA quantitative staining method for anti-slide-off cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] A method for quantitative staining of cell DNA in anti-falling sheets, comprising the following steps:

[0064] I, film production: gradient centrifugal sedimentation method, the steps are as follows:

[0065] (1) The sample is confirmed and numbered accordingly.

[0066] (2) Place the sample bottle on the shaker and shake it fully for 5 minutes.

[0067] (3) Add 2.5 ml of separation liquid (Hunan Pinsheng Biotechnology Co., Ltd., sample gradient separation liquid) into the settling chamber along the wall of the settling chamber or the wall of the filter cup.

[0068] (4) Draw 2.5ml of sample-preservation solution (Hunan Pinsheng Biotechnology Co., Ltd., liquid-based cell preservation solution) and add it along the filter cup wall.

[0069] (5) Put the symmetrical balance into a centrifuge (Hunan Pinsheng Biotechnology Co., Ltd., gradient centrifuge 4000A), centrifuge at 1200 rpm for 3 minutes.

[0070] (6) Remove the filter screen, discard the supernatant, dry it ge...

Embodiment 2

[0090] A method for quantitative staining of cell DNA in anti-falling sheets, comprising the following steps:

[0091] I, film production: gradient centrifugal sedimentation method, the steps are as follows:

[0092] (1) The sample is confirmed and numbered accordingly.

[0093] (2) Place the sample bottle on the shaker and shake it fully for 3 minutes.

[0094] (3) Add 2 ml of separation solution (Hunan Pinsheng Biotechnology Co., Ltd., sample gradient separation solution) into the settling chamber along the wall of the settling chamber or the wall of the filter cup.

[0095] (4) Draw 2ml of sample-preservation solution (Hunan Pisen Biotechnology Co., Ltd., liquid-based cell preservation solution) and add it along the wall of the filter cup.

[0096] (5) Put the symmetrical balance into a centrifuge (Hunan Pinsheng Biotechnology Co., Ltd., gradient centrifuge 4000A), 800 rpm, and centrifuge for 5 minutes.

[0097] (6) Remove the filter screen, discard the supernatant, dry ...

Embodiment 3

[0117] A method for quantitative staining of cell DNA in anti-falling sheets, comprising the following steps:

[0118] I, film production: gradient centrifugal sedimentation method, the steps are as follows:

[0119] (1) The sample is confirmed and numbered accordingly.

[0120] (2) Place the sample bottle on the shaker and shake it fully for 4 minutes.

[0121] (3) Add 1 ml of separation solution (Hunan Pinsheng Biotechnology Co., Ltd., sample gradient separation solution) into the settling chamber along the wall of the settling chamber or the wall of the filter cup.

[0122] (4) Draw 1ml of sample-preservation solution (Hunan Pinsheng Biotechnology Co., Ltd., liquid-based cell preservation solution) and add it along the wall of the filter cup.

[0123] (5) Put the symmetrical balance into a centrifuge (Hunan Pinsheng Biotechnology Co., Ltd., gradient centrifuge 4000A), centrifuge at 2000 rpm for 1 min.

[0124] (6) Remove the filter screen, discard the supernatant, dry it g...

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Abstract

The invention provides a DNA quantitative staining method for anti-slide-off cells. The method comprises the following steps: selecting an acidolysis-alkali combination mode, forming bluish violet quinone compounds in situ in a one-to-one correspondence manner, providing the maximum absorption peak in an acidic aqueous solution with the wavelength of 590 +/-15nm, and determining the absorbance toobtain the DNA content or ploidy in the cell nucleus so as to judge the physiological state and pathological change of the cell. Reagent residues are avoided through water washing, and influence of acid-base alcohol on the glass slide adhesion layer is reduced; moisture residue is avoided through baking, and the situation that hydrogen bonds are formed between water and the glass slide to reduce the adsorption capacity of the glass slide to target cells and tissues is avoided; and through operations of decoloration, dehydration and drying, the mutual vibration among molecules is accelerated, the formation of hydrogen bonds, namely the tight adhesion of cells, tissues and the glass slide, is promoted, the glass slide is prevented from falling off, and the completeness of a sampled specimenresult is guaranteed.

Description

technical field [0001] The invention relates to a staining method for quantitative detection of cell DNA, in particular to a quantitative staining method for preventing cell DNA from falling off. Background technique [0002] Deoxyribonucleic acid (DNA) staining methods include Feulgen method, methyl green-pyronin method, acridine orange fluorescence method, etc., the most classic of which is Feulgen method. Feulgen staining is a kind of cell nucleus discovered by Feulgen and Rossenbeck in 1924. IntraDNA staining technique. This method is a classic enzymatic histochemical method. The principle of Leagene Feulgen stain is that after the DNA is hydrolyzed by weak acid (1mol / L HCl), the glycosidic bond between the purine base and the deoxyribose is opened, and the phospholipid bond between the deoxyribose and the phosphoric acid is broken, forming at one end of the deoxyribose free aldehyde groups. The aldehyde group is combined with Schiff (leuco-magenta sulfite solution) r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N1/30C12Q1/68
CPCG01N1/28G01N1/30C12Q1/68G01N2001/302G01N2001/305
Inventor 贺权源刘朝前谢凯
Owner 湖南品胜生物技术有限公司
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