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Cytological DNA (Deoxyribonucleic Acid) ploidy staining kit and application

A dyeing reagent and cytology technology, applied in the biological field, can solve the problems such as the inability to accurately calculate the DNA ploidy content, the unclear demarcation of the nucleus and nucleolus, and the unclear structure.

Inactive Publication Date: 2018-09-07
南京福怡科技发展股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Pap staining is commonly used in cytology. Hematoxylin stains the nucleus with blue-purple, but the hematoxylin stains the skeleton protein in the nucleus, and it is impossible to accurately calculate the content of DNA ploidy; in the past, hematoxylin staining was used , although basophilic granules in the cytoplasm can be observed, but the structure is not clear, and the boundary between the nucleus and the nucleolus is not clear; when thionin is used instead of hematoxylin staining, the structural features of the nucleus are displayed more clearly and can be quantified accurately analysis, the sensitivity and specificity of detection are significantly improved

Method used

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  • Cytological DNA (Deoxyribonucleic Acid) ploidy staining kit and application
  • Cytological DNA (Deoxyribonucleic Acid) ploidy staining kit and application
  • Cytological DNA (Deoxyribonucleic Acid) ploidy staining kit and application

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Embodiment 1

[0026] In the staining kit of this embodiment, the components of each liter of DNA fragment mounting solution are: methanol 750ml, formaldehyde 200ml, and acetic acid 50ml; the components of each liter of DNA hydrolysis solution are: hydrochloric acid 450ml, Tris salt buffer 100ml, purified Water 450ml; the components of each liter of DNA ploidy staining solution are: thionine 0.9g, dilute sulfuric acid 2ml, sodium bisulfite powder 4g, and purified water 998ml.

[0027] The process of using the dyeing solution in this embodiment is as follows:

[0028] 1) After the cytology slide is completed, wet fix for 15 minutes;

[0029] 2) Wash with running water for 30 seconds, and use the DNA fragment mounting solution for cell fixation for 30 minutes;

[0030] 3) Wash under running water for 30 seconds, and use the DNA hydrolyzate for acid hydrolysis for 45 minutes;

[0031] 4) Wash with running water for 30 seconds, and use the DNA ploidy staining solution to stain the nucleus for 50 minutes;...

Embodiment 2

[0036] In the staining kit in this embodiment, the components of each liter of DNA fragment mounting solution are: methanol 600ml, formaldehyde 300ml, and acetic acid 100ml; the components of each liter of DNA hydrolysis solution are: sulfuric acid 500ml, phosphate buffer 300ml, purification Water 200ml; the components of each liter of DNA ploidy staining solution are: thionine 1.5g, hydrochloric acid 5ml, aluminum potassium sulfate 2g, purified water 995ml.

[0037] The process of using the dyeing solution in this embodiment is as follows:

[0038] 1) After the cytology slide is completed, wet fix for 20 minutes;

[0039] 2) Wash with running water for 30 seconds, and use the DNA fragment mounting solution for cell fixation for 40 minutes;

[0040] 3) Wash with running water for 30 seconds, and use the DNA hydrolyzate to perform acid hydrolysis for 35 minutes;

[0041] 4) Wash with running water for 30 seconds, and use the DNA ploidy staining solution to stain the nucleus for 45 minut...

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Abstract

The invention relates to a cytological DNA (Deoxyribonucleic Acid) ploidy staining kit and application and in particular relates to a staining kit containing a DNA segment sealing and fixing solution,DNA hydrolysate and DNA ploidy staining liquid and application. The staining liquid provided by the invention can meet fixation, hydrolysis and nucleus staining of a cytological DNA detection sample;an application method is simple and convenient, a staining effect is stable and uniform and a background is clean and clear; the staining liquid can be accurately identified through a digitalized scanner; the validity period of the staining liquid at room temperature is one year or more.

Description

Technical field [0001] The present invention relates to the field of biotechnology, in particular to a cytological DNA ploidy staining box and application. Background technique [0002] Analyzing the cell cycle and DNA ploidy of cells has been increasingly used in clinics, providing an important basis for tumor diagnosis, efficacy evaluation and prognosis prediction. It is also increasingly used in cell dynamics and cell regulation in scientific research. Death observation and other aspects. DNA ploidy analysis combined with clinical pathological morphological diagnosis, early diagnosis of some malignant tumors, follow-up follow-up and early treatment, improve the cure rate and survival rate of tumors. Especially for the analysis of cell extracts, body fluids, glandular secretions, and tissue exfoliated cells, the significance is very important. DNA ploidy analysis technology is of great value in the early diagnosis and prognosis of bladder cancer, prostate cancer, ovarian canc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/30
CPCG01N1/30
Inventor 姚斌左露露
Owner 南京福怡科技发展股份有限公司