CN107367417B - Rapid fixing agent composition for punctured cells under ultrasonic guidance and preparation method thereof - Google Patents
Rapid fixing agent composition for punctured cells under ultrasonic guidance and preparation method thereof Download PDFInfo
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- CN107367417B CN107367417B CN201710574110.2A CN201710574110A CN107367417B CN 107367417 B CN107367417 B CN 107367417B CN 201710574110 A CN201710574110 A CN 201710574110A CN 107367417 B CN107367417 B CN 107367417B
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- Prior art keywords
- fixing agent
- agent
- cells
- punctured
- hydrochloric acid
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- 239000000203 mixture Substances 0.000 title claims abstract description 17
- 238000002360 preparation method Methods 0.000 title abstract description 5
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 22
- 238000002604 ultrasonography Methods 0.000 claims abstract description 8
- 239000003219 hemolytic agent Substances 0.000 claims abstract description 7
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 38
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 38
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 37
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 36
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 26
- 239000001103 potassium chloride Substances 0.000 claims description 19
- 235000011164 potassium chloride Nutrition 0.000 claims description 19
- 239000011780 sodium chloride Substances 0.000 claims description 19
- 238000003756 stirring Methods 0.000 claims description 14
- 229960000583 acetic acid Drugs 0.000 claims description 13
- 239000012362 glacial acetic acid Substances 0.000 claims description 13
- 239000000843 powder Substances 0.000 claims description 7
- 239000000834 fixative Substances 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 210000004027 cell Anatomy 0.000 abstract description 37
- 238000003745 diagnosis Methods 0.000 abstract description 11
- 210000003743 erythrocyte Anatomy 0.000 abstract description 7
- 230000035699 permeability Effects 0.000 abstract 1
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 238000010186 staining Methods 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- 210000003855 cell nucleus Anatomy 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/305—Fixative compositions
Abstract
The invention belongs to the technical field of cell engineering, and relates to a rapid fixing agent composition for punctured cells under ultrasonic guidance and a preparation method thereof. A composition of a rapid fixing agent for punctured cells under the guidance of ultrasound is characterized by comprising the following components in percentage by weight: cell fixing agent: 60-95%; low permeability agent: 0.2-4%; hemolytic agent: 10 to 25 percent. The composition of the rapid fixing agent for the punctured cells has the advantages of maximally keeping the shape of target cells, and maximally eliminating the covering of the target cells by background red blood cells, so that diagnosis can be made in the shortest time and the diagnosis accuracy is improved.
Description
Technical Field
The invention belongs to the technical field of cell engineering, and relates to a rapid fixing agent composition for punctured cells under ultrasonic guidance and a preparation method thereof.
Background
In the field of punctured cell staining detection technology, 95% ethanol fixation methods that last decades are currently used by most medical institutions. The fixing is carried out for more than 10 minutes according to the routine requirement, and with the rapid development of new clinical techniques, such as puncture cytology diagnosis under the guidance of ultrasonic waves, the clinical requirement is that the information of whether the target cells (tumor cells) are punctured can be obtained immediately on a puncture site, and the quality and the malignancy of the cells can be accurately judged.
Because cell puncture under the guidance of ultrasound needs to be carried out in ultrasound or clinic with professional equipment, a puncture smear is generally required to be sent to be detected in a pathology department or a clinical laboratory for cell diagnosis, and thus, a patient needs to wait for a long time under anesthesia in puncture of some parts, and accidents are easy to happen; in addition, the punctured cells cannot be fixed and stained in time, and the accurate diagnosis can be affected.
Therefore, the known rapid fixing agent for punctured cells has various inconveniences and problems as described above.
Disclosure of Invention
The invention aims to provide a rapid fixing agent composition for punctured cells under the guidance of ultrasound and a preparation method thereof.
Another object of the present invention is to allow rapid and accurate diagnosis in paracentesis cytology.
In order to achieve the purpose, the invention is realized by the following technical scheme:
an ultrasonic-guided punctured cell rapid fixative composition comprises the following components in percentage by weight:
60-95% of cell fixing agent
0.1 to 5 percent of low-permeability agent
5-35% of hemolytic agent.
Further, the rapid fixing agent composition for the punctured cells under the guidance of the ultrasound comprises the following components in percentage by weight:
70-95% of cell fixing agent
0.2 to 4 percent of low-permeability agent
10-25% of hemolytic agent.
The cell fixing agent is ethanol, and the concentration of the ethanol is 15-98%.
The low-permeability agent is a mixture of sodium chloride and potassium chloride, and the weight ratio of the sodium chloride to the potassium chloride is (1-20): 1 to 20.
The hemolytic agent is one or a mixture of hydrochloric acid and glacial acetic acid, and the content of the hydrochloric acid is 36-38%.
The purity of the sodium chloride is 99.5-100%, and the purity of the potassium chloride is 99.7-100%.
A method for preparing a rapid fixative composition for punctured cells under the guidance of ultrasound, comprising the following steps:
(1) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(2) and (3) adding hydrochloric acid and glacial acetic acid into the solution obtained in the step (1), and uniformly stirring.
The invention has the beneficial effects that:
(1) the composition of the rapid fixing agent for punctured cells has the advantage of maximally keeping the shape of target cells.
(2) The invention can achieve rapidness and accuracy in puncture cytology diagnosis.
(3) The invention can eliminate the red blood cell background on the smear while fixing.
Detailed Description
The invention will now be further illustrated by reference to the following examples:
example 1
(1) Weighing 80 ml of 50% ethanol, 20 ml of 38% hydrochloric acid, 0.3 g of sodium chloride and 0.5 g of potassium chloride;
(2) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(3) and (3) adding hydrochloric acid into the solution obtained in the step (2), and uniformly stirring to obtain a reagent 1.
Example 2
(1) Weighing 80 ml of 50% ethanol, 20 ml of 38% hydrochloric acid, 0.3 g of sodium chloride and 0.5 g of potassium chloride;
(2) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(3) and (3) adding hydrochloric acid into the solution obtained in the step (2), and uniformly stirring to obtain a reagent 2.
Example 3
(1) Weighing 80 ml of 50% ethanol, 15 ml of 38% hydrochloric acid, 5 ml of glacial acetic acid, 0.3 g of sodium chloride and 0.5 g of potassium chloride;
(2) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(3) and (3) adding hydrochloric acid and glacial acetic acid into the solution obtained in the step (2), and uniformly stirring to obtain a reagent 3.
Example 4
(1) Weighing 80 ml of 50% ethanol, 20 ml of glacial acetic acid, 0.3 g of sodium chloride and 0.5 g of potassium chloride;
(2) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(3) adding glacial acetic acid into the solution obtained in the step 2, and uniformly stirring to obtain a reagent 4.
Example 5
(1) Weighing 80 ml of 50% ethanol, 5 ml of 38% hydrochloric acid, 15 ml of glacial acetic acid, 0.3 g of sodium chloride and 0.5 g of potassium chloride;
(2) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(3) and (3) adding hydrochloric acid into the solution obtained in the step (2), and uniformly stirring to obtain a reagent 5.
And (3) carrying out an ultrasonic guided puncture cell fixation experiment on the obtained reagents 1-5, and using a push sheet method and carrying out fixed staining according to the currently universal 95% alcohol as a control group. The specific test results are shown in table 1 below.
TABLE 1
In the cell morphology in the above table:
0 shows essentially no cell structure;
+ cells do not shrink significantly;
+ cells had visible contraction;
the cells contract and deform.
In the staining results:
the cell nucleus and the serous staining are not clear;
the nucleus and the pulp dyeing can be distinguished;
the cell nucleus and pulp staining are bright and clear, and the contrast is clear.
In smear background:
almost no coverage of the formed red blood cells in the background, the cells of interest can be found quickly and the diagnosis is made unequivocal.
+ a small number of formed erythrocytes in the background, no target cells covered, and a clear diagnosis.
The positive and negative background is covered by formed red blood cells, which can cover the target cells to a certain extent and affect the diagnosis.
A large number of shaped red blood cells are accumulated in the background of +++, covering the target cells, and affecting the diagnosis.
The experimental results show that the embodiment of the invention can obviously reduce the existence of red blood cells in the background, and has small influence on the cells, the reagent No. 5 contains 80 ml of 50% ethanol, 5 ml of 38% hydrochloric acid, 15 ml of glacial acetic acid, 0.3 g of sodium chloride and 0.5 g of potassium chloride, and the effect is optimal, which indicates that the fixative achieves better effect when hydrochloric acid and glacial acetic acid exist in the fixative at the same time and the content of the glacial acetic acid is greater than that of the hydrochloric acid.
The above embodiments are provided only for illustrating the present invention and not for limiting the present invention, and those skilled in the art can make various changes or modifications without departing from the spirit and scope of the present invention. Accordingly, all equivalents are intended to fall within the scope of the invention, which is defined in the claims.
Claims (1)
1. The rapid fixing agent composition for the punctured cells under the guidance of ultrasound is characterized by comprising the following components in percentage by weight: 70-95% of cell fixing agent
0.2 to 4 percent of low-permeability agent
10-25% of hemolytic agent;
the cell fixing agent is ethanol, and the concentration of the ethanol is 15-98%;
the low-permeability agent is a mixture of sodium chloride and potassium chloride, and the weight ratio of the sodium chloride to the potassium chloride is (1-20): 1-20;
the hemolytic agent is a mixture of hydrochloric acid and glacial acetic acid, and the concentration of the hydrochloric acid is 36-38%; the content of glacial acetic acid in the hemolytic agent is greater than the content of hydrochloric acid;
the purity of the sodium chloride is 99.5-100%, and the purity of the potassium chloride is 99.7-100%;
a method for puncturing a cell rapid fixative composition under the guidance of ultrasound, comprising the steps of:
(1) adding sodium chloride and potassium chloride powder into prepared ethanol with concentration under stirring for dissolving;
(2) and (3) adding hydrochloric acid and glacial acetic acid into the solution obtained in the step (1), and uniformly stirring.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710574110.2A CN107367417B (en) | 2017-07-14 | 2017-07-14 | Rapid fixing agent composition for punctured cells under ultrasonic guidance and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710574110.2A CN107367417B (en) | 2017-07-14 | 2017-07-14 | Rapid fixing agent composition for punctured cells under ultrasonic guidance and preparation method thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107367417A CN107367417A (en) | 2017-11-21 |
| CN107367417B true CN107367417B (en) | 2021-06-22 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710574110.2A Active CN107367417B (en) | 2017-07-14 | 2017-07-14 | Rapid fixing agent composition for punctured cells under ultrasonic guidance and preparation method thereof |
Country Status (1)
| Country | Link |
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| CN (1) | CN107367417B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182506A (en) * | 2007-11-07 | 2008-05-21 | 曾思恩 | Fixation fluid for saving cast-off cells and manufacturing method of cell shallow layer smear |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| PT1924252E (en) * | 2005-07-28 | 2011-12-16 | Id Fish Technology Inc | Method for improving cell permeability to foreign particles |
| CN101246098B (en) * | 2008-03-14 | 2010-08-18 | 成都大学 | Horse bean root tip cell microkernel flaking method |
| CN102313662A (en) * | 2010-06-30 | 2012-01-11 | 上海市静安区青少年活动中心 | Application of Roselle pigment in dyeing of chromosome |
| CN102147417B (en) * | 2011-01-14 | 2013-11-06 | 中国农业大学 | Method for positioning immune tissues of growth hormone for malus plants and application thereof |
| PT2705345E (en) * | 2011-05-02 | 2015-06-17 | Lapenna José Carlos | Use of halogeno-cyanoacetamides for fixation and preservation of biological samples |
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- 2017-07-14 CN CN201710574110.2A patent/CN107367417B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182506A (en) * | 2007-11-07 | 2008-05-21 | 曾思恩 | Fixation fluid for saving cast-off cells and manufacturing method of cell shallow layer smear |
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| CN107367417A (en) | 2017-11-21 |
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Inventor after: Qiu Chengmin Inventor after: Zhang Li Inventor after: Ren Dabin Inventor after: Gu Minmin Inventor after: Qian Xiuhong Inventor before: Qiu Chengmin Inventor before: Zhang Li |
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