CN114047050A - Sample diluent for cell embedding machine and preparation method thereof - Google Patents

Abstract

The invention belongs to the technical field of pathological cell embedding, and particularly relates to a sample diluent for a cell embedding machine and a preparation method thereof, which are characterized by comprising a composition of a PH buffering agent, agar and a preservative, wherein the PBS buffering agent is 100ml, the agar is 1-4g, and the preservative is 1-3 ml. The pH of the PBS buffer was 6.86. The content of the preservative is 40% of formaldehyde solution. The cell type classification method has extremely high encapsulation performance, enables cells to be tightly gathered into cell blocks, has clean slicing background and no impurities, is tightly gathered and not dispersed, is visual and convenient for a pathologist to judge the properties of the cells, can well classify the cell types when searching the tumor cells, and provides more reference basis for the standard treatment of tumors.

Description

Sample diluent for cell embedding machine and preparation method thereof

Technical Field

The invention belongs to the technical field of pathological cell embedding, and particularly relates to a sample diluent for a cell embedding machine and a preparation method thereof.

Background

In pathological research, the cell wax block is mainly used for long-term preservation of cytological samples, including alveolar lavage fluid, cerebrospinal fluid, hydrothorax and ascites, pericardial fluid, urine, various puncture samples and the like, and can be used for subsequent diagnosis and verification and development of cytological immunohistochemical experiments. At present, the manual preparation of the cell wax block needs repeated pre-centrifugation and takes long time, and the obtained cell sediment is easy to disperse or lose in the dehydration process. For samples with small cell amount such as alveolar lavage fluid, cerebrospinal fluid, urine, puncture and the like, most of the common methods cannot be used for preparing the samples meeting the clinical requirements.

The cell embedding machine equipment can gather trace cells, the cells are tightly gathered into cell blocks, the background of the slice is clean and free of impurities, the cells are tightly gathered and are not dispersed, so that the cell embedding machine equipment is visually convenient for a pathologist to judge the properties of the cells, and when the tumor cells are searched, the cell types can be well classified, so that more reference bases are provided for the standard treatment of tumors. Compared with the prior ordinary smear of the exfoliative cytology, the technology has higher accuracy, more convenient storage and more definite diagnosis of diseases. And the kit can be continuously used for detection projects such as immunohistochemistry, FISH and the like, and has clinical value for later-stage research analysis and preservation. After the cell embedding preparation, N antibodies can be used for performing immunohistochemical work on N types of cells.

Disclosure of Invention

In order to make up for the defects of the prior art, the invention provides a sample diluent for a cell embedding machine and a preparation method thereof.

The sample diluent for the cell embedding machine is characterized by comprising a composition of a pH buffer, an osmotic pressure maintaining agent and a preservative, wherein the pH buffer is 100ml, the agar is 1-4g, and the preservative is 1-3 ml.

The sample diluent for the cell embedding machine is characterized in that the pH value of the pH buffer is 6.86.

The sample diluent for the cell embedding medium is characterized in that the pH buffer is a PBS solution.

The sample diluent for the cell embedding machine is characterized in that the preservative is formaldehyde solution with 35-40% of formaldehyde content.

A preparation method of a sample diluent for a cell embedding machine is characterized by comprising the following steps:

s1 adding agar 1-4g into pH buffer (100 ml) with pH of 6.86 at room temperature;

s2, stirring at constant speed continuously until the agar is melted;

s3 finally adding a preservative.

The preparation method of the sample diluent for the cell embedding medium is characterized in that in the step S1, the PH buffer is a normal-temperature PBS solution;

in the step S2, stirring is performed by using a stirring rod, wherein the stirring speed is 120-150 rpm;

the preservative in step S3 is a 40% formaldehyde solution.

Compared with the prior art, the invention has the following advantages:

the sample diluent for the cell embedding machine has extremely high encapsulation property in the cell block manufacturing process, enables cells to be tightly aggregated into cell blocks, has clean slicing background and no impurities, is tightly aggregated and not dispersed, is visual and convenient for a pathologist to judge the properties of the cells, can well classify the cell types when searching the tumor cells, and provides more reference basis for the standard treatment of tumors. The qualification rate of cell block preparation is high, and the accuracy of pathology research can be improved. Meanwhile, the energy of medical staff or scientific research personnel in the process of manufacturing the cell block can be reduced.

Detailed Description

A sample diluent for a cell embedding machine comprises a PBS buffer solution, an osmotic pressure maintaining agent and a preservative, wherein the pH buffer solution is 100ml, the agar is 1-4g, and the preservative is 1-3 ml.

The pH value of the pH buffering agent is 6.86, and the pH buffering agent is PBS solution.

The formaldehyde content of the preservative is 35-40% of formaldehyde aqueous solution.

Example 1

A sample diluent for a cell embedding machine is a combination of 100ml of pH buffer with pH 6.86, 1g of agar and 1ml of preservative, the preservative is 35% formaldehyde solution, and the pH buffer is PBS solution.

Example 2

A sample diluent for a cell embedding machine is a combination of 100ml of pH buffer with pH 6.86, 2.5g of agar and 2ml of preservative, the preservative is 40% formaldehyde solution, and the pH buffer is PBS solution.

Example 3

A sample diluent for a cell embedding machine is a combination of 100ml of pH buffer with pH 6.86, 4g of agar and 2ml of preservative, the preservative is 40% formaldehyde solution, and the pH buffer is PBS solution.

At normal temperature, the sample diluent is a jelly-like solid, when a cell block for pathological research is manufactured, the sample diluent needs to be heated firstly, cells are added into a centrifugal tube containing the sample diluent after being melted, centrifugation is carried out after the cells are fully and uniformly mixed, the cells are completely wrapped by the sample diluent, the cells are deposited at the bottom of the sample diluent, and the cells are taken out after being cooled and then are subjected to next embedding treatment.

A preparation method of a sample diluent for a cell embedding machine comprises the following steps: adding agar 1-4g into pH buffer with pH of 6.86 at room temperature, wherein the pH buffer is 100 ml; then stirring at constant speed continuously until the agar is melted; finally, a preservative is added.

The pH buffer is normal temperature PBS solution. During stirring, the stirring was continued at a stirring speed of 120-. After stirring was complete, 40% formaldehyde solution was added.

The conventional sample diluent and the sample diluent according to each set of examples of the present invention were used for the cell mass preparation process, and the test results are shown in the following table. When the cell mass is qualified, the cells aggregate at the lowest end and there are no discrete cells in the sample diluent.

Experimental group	Number of the standard example	Number of qualified products	Percent of pass
				Traditional sample diluent	100	90	80％
Example 1	100	99	99％
				Example 2	100	100	100％
Example 3	100	100	100％

The experimental data in the table are combined to show that the sample diluent can be better fused with cells compared with the traditional sample diluent, the cell morphology is protected, and the cell block manufacturing yield is high.

The sample diluent of the present invention was used to perform the cell block preparation experiment in different units, and the experimental results are shown in the following table.

According to the experimental data surface, the sample diluent for the cell embedding machine has extremely high wrapping property in the cell block manufacturing process, so that cells are tightly gathered into a cell block, the background of a section is clean and free of impurities, the cells are tightly gathered and are not dispersed, the property of the cells can be visually judged by a pathologist conveniently, when tumor cells are found, the cell types can be well classified, and more reference basis is provided for the standard treatment of tumors. The qualification rate of cell block preparation is high, and the accuracy of pathology research can be improved. Meanwhile, the energy of medical staff or scientific research personnel in the process of manufacturing the cell block can be reduced.

Finally, it should be noted that: the above embodiments are only used to illustrate the technical solution of the present invention, and not to limit the same; while the invention has been described in detail and with reference to the foregoing embodiments, it will be understood by those skilled in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some or all of the technical features may be equivalently replaced; and the modifications or the substitutions do not make the essence of the corresponding technical solutions depart from the scope of the technical solutions of the embodiments of the present invention.

Claims (6)

1. A sample diluent for a cell embedding machine, characterized by comprising a composition of a pH buffer, an osmotic pressure maintaining agent and a preservative, wherein the pH buffer is 100ml, the agar is 1-4g, and the preservative is 1-3 ml.

2. The specimen dilution liquid for a cell-embedding machine according to claim 1, wherein the pH buffer has a pH of 6.86.

3. The sample diluent for a cell embedding medium according to claim 2, characterized in that the PH buffer is a PBS solution.

4. The specimen dilution liquid for a cell embedding machine according to claim 1, characterized in that the preservative is a formaldehyde solution having a formaldehyde content of 35% to 40%.

5. A method for preparing a sample diluent for a cell embedding machine according to any one of claims 1 to 4, characterized by comprising the steps of:

s1 adding agar 1-4g into pH buffer (100 ml) with pH of 6.86 at room temperature;

s2, stirring at constant speed continuously until the agar is melted;

s3 finally adding a preservative.

6. The method for preparing a sample diluent for a cell embedding medium according to claim 5, wherein in the step S1, the pH buffer is a normal temperature PBS solution;

in the step S2, stirring is performed by using a stirring rod, wherein the stirring speed is 120-150 rpm;

the preservative in step S3 is a 40% formaldehyde solution.