阅读说明：本技术 一种液基细胞保存液及其制备方法 (Liquid-based cell preservation solution and preparation method thereof ) 是由 刘益民 卞忠力 徐安双 于 2019-11-27 设计创作，主要内容包括：本发明公开了一种液基细胞保存液,其成分包括：酒精300-350ml,水600-650ml,磷酸二氢钾15g,磷酸氢二钠8g,乙二胺四乙酸二钠1.8g,二硫苏糖醇1.5g,氯化铵0.9g,叠氮钠0.2g；本发明还提供了上述液基细胞保存液的制备方法；该液基细胞保存液够快速固定细胞,稳定细胞的PH值,防止标本中血液凝结,及时裂解标本中的红细胞,溶解标本中粘液的粘蛋白,同时防腐杀菌,使标本可以长期保存。(The invention discloses a liquid-based cell preservation solution, which comprises the following components: 350ml of alcohol, 600ml of water, 650ml of monopotassium phosphate, 8g of disodium hydrogen phosphate, 1.8g of ethylene diamine tetraacetic acid, 1.5g of dithiothreitol, 0.9g of ammonium chloride and 0.2g of sodium azide; the invention also provides a preparation method of the liquid-based cell preservation liquid; the liquid-based cell preservation liquid can quickly fix cells, stabilize the pH value of the cells, prevent blood coagulation in a specimen, timely lyse red blood cells in the specimen, dissolve mucin in the specimen, and simultaneously prevent corrosion and sterilize, so that the specimen can be preserved for a long time.)

1. A liquid-based cell preservation solution, which is characterized in that: the liquid-based cell preservation liquid comprises the following components: 350ml of alcohol 300-.

2. The fluid-based cell preservation fluid according to claim 1, wherein: the pH value of the liquid-based cell preservation solution is 5.2-6.3.

3. The fluid-based cell preservation fluid according to claim 1, wherein: the purity of the alcohol is 99.99%.

4. The fluid-based cell preservation fluid according to claim 1, wherein: the water is deionized water.

5. A preparation method of liquid-based cell preservation solution is characterized by comprising the following steps: the preparation method comprises the following preparation steps:

a. dissolving potassium dihydrogen phosphate and disodium hydrogen phosphate in a formula amount in 100ml of deionized water, and uniformly stirring to obtain a buffer solution a;

b. dissolving the disodium ethylene diamine tetraacetate, dithiothreitol, ammonium chloride and sodium azide in the formula ratio in the buffer solution a, and uniformly stirring to obtain a solution b;

c. slowly adding the rest amount of deionized water into the obtained solution b, and stirring uniformly while adding to obtain a solution c;

d. slowly adding the alcohol with the formula amount into the solution c, and stirring uniformly while adding.

Technical Field

The invention relates to the technical field of pathological examination, in particular to liquid-based cell preservation liquid used in pathological examination of cervical exfoliated cells of a human body and a preparation method thereof.

Background

The liquid-based cytology examination adopts a liquid-based thin-layer cell detection system to detect cervical cells and carry out cytological classification diagnosis, is a more advanced cervical cancer cytology examination technology internationally at present, obviously improves the satisfaction degree of specimens and the abnormal cell detection rate of the cervix compared with the traditional cervical smear examination by scraping and smearing, has the detection rate of the cervical cancer prevention cytology examination on the cervical cancer cells of 100 percent, and can discover partial precancerous lesions and microbial infections such as mould, trichomonas, virus and chlamydia at the same time.

The liquid-based thin-layer cell detection method obviously improves the detection quality of cervical cell samples and becomes a main detection method on the market, liquid-based cell preservation solution is one of the key links of the whole liquid-based thin-layer cell detection method, once cells are separated from the original living environment, the morphological structure and the surface characteristics of the cells can be changed, so that the original characteristics of the cells can not be changed only by a correct cell preservation method, and the variable of cell instability in some important experiments can be eliminated, so that the stability and repeatability of cell-related experiments are greatly improved, the components of exfoliated cells of human cervix are complex and often contain mucus and blood, and the cell preservation solution needs to fix useful components in a specimen and eliminate useless components so as to avoid interference on experimental results.

At present, a plurality of commercially available liquid-based cell preservation solutions are clinically applied, represented by new cupra (Cytye, Boxborough, MA, currently called Hologic) approved by FDA in the usa in 1996 and Autocyte Prep (currently called SurePath, TripPath Imaging, Burlington, NC, currently called BD diagnosis company) approved in 1999, the two imported cell preservation solutions are expensive to prepare and are used together with a matched machine, and it is difficult for some small and medium hospitals to have use conditions, and the cells after the cell preservation solution is stored have different preservation solution components, different PH values, different protein treatments, cell adhesion to an adhesive slide, staining of staining solution, influence preservation of cell solution, and related to subsequent specimen preparation, and the preservation solution appearing in the market often has no requirement for subsequent specimen treatment.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide the liquid-based cell preservation solution and the preparation method thereof, which can better preserve cervical exfoliated cells, protect the integrity and the fixity of the cells, avoid the solid shrinkage, the dissolution and the expansion of the cells, decompose red blood cells, disperse mucus, remove interference components and provide a good foundation for subsequent slice-making and dyeing.

In order to achieve the purpose, the invention provides the following technical scheme: the liquid-based cell preservation solution is characterized by comprising the following components: 350ml of alcohol 300-.

Further, the pH value of the liquid-based cell preservation solution is 5.2-6.3.

Further, the purity of alcohol was 99.99%.

Further, the water is deionized water.

A preparation method of liquid-based cell preservation solution comprises the following steps:

a. dissolving potassium dihydrogen phosphate and disodium hydrogen phosphate in a formula amount in 100ml of deionized water, and uniformly stirring to obtain a buffer solution a;

b. dissolving the disodium ethylene diamine tetraacetate, dithiothreitol, ammonium chloride and sodium azide in the formula ratio in the buffer solution a, and uniformly stirring to obtain a solution b;

c. slowly adding the rest amount of deionized water into the obtained solution b, and stirring uniformly while adding to obtain a solution c;

d. slowly adding the alcohol with the formula amount into the solution c while stirring uniformly;

all the steps are carried out at normal temperature and normal pressure.

Compared with the prior art, the invention has the beneficial effects that: the liquid-based cell preservation solution provided by the invention has comprehensive functions, and adopts alcohol as a fixing agent of the liquid-based cell preservation solution to quickly and effectively fix the liquid-based cells in the specimen; potassium dihydrogen phosphate and disodium hydrogen phosphate are used as acid-base buffer solution of the liquid-based cell preservation solution, when the pH value of the liquid-based cell preservation solution is too high, redundant alkali and dihydrogen phosphate ions form hydrogen phosphate ions, the pH value of the solution is reduced, and when the pH value is too low, the hydrogen phosphate ions and hydrogen ions in acid form dihydrogen phosphate ions, so that the pH value of the solution is improved, the constant pH value of the solution is favorably maintained, and cells cannot be broken due to pH change in cell fixation; disodium ethylene diamine tetraacetate is used as an anticoagulant of the liquid-based cell preservation solution, so that blood in a specimen is prevented from being coagulated into blocks when the liquid-based cell preservation solution fixes cells; adopting dithiothreitol as a mucolytic agent of the liquid-based cell preservation solution to dissolve mucin in the specimen; adopting ammonium chloride as hemolytic agent of liquid-based cell preservation solution to lyse erythrocytes in the specimen; sodium azide is used as a preservative of the liquid-based cell preservation solution to prevent the liquid-based cell preservation solution from deteriorating; the liquid-based cell preservation solution provided by the invention has complete functions, is multipurpose, can quickly fix cells, stabilize the pH value of the cells, prevent blood in a specimen from coagulating, immediately crack red blood cells in the specimen, dissolve mucin in mucus in the specimen, and simultaneously carry out antiseptic sterilization so that the specimen can be preserved for a long time.

Detailed Description

In the following, embodiments of the present invention will be described clearly and completely, and it is to be understood that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by those skilled in the art without any inventive work based on the embodiments of the present invention belong to the scope of the present invention.