CN110742059A - Liquid-based cell preservation solution and preparation method thereof - Google Patents
Liquid-based cell preservation solution and preparation method thereof Download PDFInfo
- Publication number
- CN110742059A CN110742059A CN201911179836.1A CN201911179836A CN110742059A CN 110742059 A CN110742059 A CN 110742059A CN 201911179836 A CN201911179836 A CN 201911179836A CN 110742059 A CN110742059 A CN 110742059A
- Authority
- CN
- China
- Prior art keywords
- liquid
- cell preservation
- based cell
- solution
- specimen
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0226—Physiologically active agents, i.e. substances affecting physiological processes of cells and tissue to be preserved, e.g. anti-oxidants or nutrients
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N1/00—Preservation of bodies of humans or animals, or parts thereof
- A01N1/02—Preservation of living parts
- A01N1/0205—Chemical aspects
- A01N1/021—Preservation or perfusion media, liquids, solids or gases used in the preservation of cells, tissue, organs or bodily fluids
- A01N1/0215—Disinfecting agents, e.g. antimicrobials for preserving living parts
Abstract
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.
Description
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.
Example one
The components of the liquid-based cell preservation solution in the embodiment comprise: 300ml of alcohol, 650ml of water, 15g 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, wherein the purity of the alcohol is 99.99%, the water is deionized water, and the pH value of the liquid-based cell preservation solution is 5.2-6.3.
The preparation method of the 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.
Example two
The components of the liquid-based cell preservation solution in the embodiment comprise: 350ml of alcohol, 600ml of water, 15g 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, wherein the purity of the alcohol is 99.99%, the water is deionized water, and the pH value of the liquid-based cell preservation solution is 5.2-6.3.
The preparation method of the 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.
The cell preservation solution has reasonable component proportion, 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; meanwhile, the preparation cost is low, and the popularization is easy.
Although the present invention has been described in detail with reference to the foregoing examples, it will be apparent to one skilled in the art that various changes, modifications and variations can be made in the embodiments described above, or equivalent arrangements of some features of the embodiments described above.
Claims (5)
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.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911179836.1A CN110742059A (en) | 2019-11-27 | 2019-11-27 | Liquid-based cell preservation solution and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911179836.1A CN110742059A (en) | 2019-11-27 | 2019-11-27 | Liquid-based cell preservation solution and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110742059A true CN110742059A (en) | 2020-02-04 |
Family
ID=69284737
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911179836.1A Pending CN110742059A (en) | 2019-11-27 | 2019-11-27 | Liquid-based cell preservation solution and preparation method thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110742059A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112640888A (en) * | 2020-12-30 | 2021-04-13 | 深路医学科技(武汉)有限公司 | Cell preservation solution, preparation method thereof and cell preservation method |
| CN113575573A (en) * | 2021-09-07 | 2021-11-02 | 易普森生物科技(深圳)有限公司 | Preservation solution for sputum sample and application thereof |
| CN113729009A (en) * | 2021-10-14 | 2021-12-03 | 山东高创医疗器械国家研究院有限公司 | Cervical cell preservation solution and preparation method and preservation method thereof |
| CN114149960A (en) * | 2021-12-03 | 2022-03-08 | 山东高创医疗器械国家研究院有限公司 | Sample density separating medium and cell separating method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105409925A (en) * | 2014-09-19 | 2016-03-23 | 孝感宏翔生物医械技术有限公司 | Liquid-based cell preservation solution |
| WO2017002861A1 (en) * | 2015-06-30 | 2017-01-05 | シスメックス株式会社 | Cell preservative solution, use of same, and method for producing cell preservative solution |
| CN107410287A (en) * | 2017-08-17 | 2017-12-01 | 广州鸿琪光学仪器科技有限公司 | Cell-preservation liquid and its application |
| CN109682971A (en) * | 2018-12-18 | 2019-04-26 | 杭州海世嘉生物科技有限公司 | A kind of preservation liquid and its preparation process, application method |
-
2019
- 2019-11-27 CN CN201911179836.1A patent/CN110742059A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105409925A (en) * | 2014-09-19 | 2016-03-23 | 孝感宏翔生物医械技术有限公司 | Liquid-based cell preservation solution |
| WO2017002861A1 (en) * | 2015-06-30 | 2017-01-05 | シスメックス株式会社 | Cell preservative solution, use of same, and method for producing cell preservative solution |
| CN107410287A (en) * | 2017-08-17 | 2017-12-01 | 广州鸿琪光学仪器科技有限公司 | Cell-preservation liquid and its application |
| CN109682971A (en) * | 2018-12-18 | 2019-04-26 | 杭州海世嘉生物科技有限公司 | A kind of preservation liquid and its preparation process, application method |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112640888A (en) * | 2020-12-30 | 2021-04-13 | 深路医学科技(武汉)有限公司 | Cell preservation solution, preparation method thereof and cell preservation method |
| CN112640888B (en) * | 2020-12-30 | 2022-05-03 | 深路医学科技(武汉)有限公司 | Cell preservation solution, preparation method thereof and cell preservation method |
| CN113575573A (en) * | 2021-09-07 | 2021-11-02 | 易普森生物科技(深圳)有限公司 | Preservation solution for sputum sample and application thereof |
| CN113729009A (en) * | 2021-10-14 | 2021-12-03 | 山东高创医疗器械国家研究院有限公司 | Cervical cell preservation solution and preparation method and preservation method thereof |
| CN114149960A (en) * | 2021-12-03 | 2022-03-08 | 山东高创医疗器械国家研究院有限公司 | Sample density separating medium and cell separating method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110742059A (en) | Liquid-based cell preservation solution and preparation method thereof | |
| CN113383769B (en) | Cell preservation solution and preparation method and application thereof | |
| CN107509722B (en) | Specimen preservation solution | |
| US7666620B2 (en) | Rapid tissue processing system | |
| CN103120153A (en) | Exfoliative cells preserving fluid | |
| CN107410287A (en) | Cell-preservation liquid and its application | |
| CN108402033A (en) | One kind having preprocessing function cervical exfoliated cell preservative fluid and application | |
| JP6142102B2 (en) | Cell preservation solution and use thereof, and method for producing cell preservation solution | |
| EP0511430A2 (en) | Cell preservative solution | |
| CN109644985B (en) | Cell preservation solution and application thereof | |
| CN113729009B (en) | Cervical cell preservation solution and preparation method and preservation method thereof | |
| CN112640888B (en) | Cell preservation solution, preparation method thereof and cell preservation method | |
| CN108849850A (en) | A kind of human body cervical exfoliated cell preservative fluid | |
| CN113966737B (en) | Cell preservation solution for in-vitro analysis and detection and preparation method thereof | |
| KR101924401B1 (en) | preserving solution for oral epithelium cell and manufacturing method thereof | |
| CN113575573A (en) | Preservation solution for sputum sample and application thereof | |
| CN115176798A (en) | Liquid-based cell preservation solution and preparation method and application thereof | |
| KR100987949B1 (en) | Cell-preserving solution | |
| CN110037011A (en) | A kind of preparation method and applications of soybean lecithin freeze-extender | |
| JP2023510252A (en) | Nucleic acid and cell preservative compositions and methods of use | |
| CN114568423A (en) | Preparation and application of multipurpose novel cell preservation solution | |
| CN107271246A (en) | A kind of immune p16 for Thinprep pap testINK4aAntigen preserves liquid and preparation method thereof | |
| CN112741080B (en) | Liquid-based cell preservation treatment liquid and preparation method and application thereof | |
| Rawlins | Cytological examination of sputum in relation to its macroscopic purulence | |
| KR101133997B1 (en) | A composite for the use of cytological analysis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200204 |
|
| WD01 | Invention patent application deemed withdrawn after publication |