CN118565950A - Dry-method dyeing vaginal secretion dyeing liquid and preparation method and dyeing method thereof - Google Patents

Abstract

The invention relates to the technical field of cell staining, in particular to a dry method vaginal secretion staining solution, a preparation method and a staining method thereof, comprising a first staining solution and a second staining solution; the first dyeing liquid comprises blue dye, basic fuchsin, an organic solvent and deionized water; the second staining solution comprises eosin, buffer solution, organic solvent and deionized water. Solves the problems of convenient operation and poor dyeing effect in the dry dyeing of vaginal secretion. Solves the problems of good dyeing effect but poor storage stability of the dyeing liquid in the dyeing of vaginal secretion. Solves the problems of good dyeing effect but batch-to-batch difference of dyeing liquid in the vaginal secretion dyeing.

Description

Dry-method dyeing vaginal secretion dyeing liquid and preparation method and dyeing method thereof

Technical Field

The invention relates to the technical field of cell staining, in particular to a dry-method vaginal secretion staining solution and a preparation method and a staining method thereof.

Background

Vaginal secretions have a composition test that has two modes, a wet test and a dry test. Wet-inspection is also two kinds of direct microscopic observation and post-staining microscopic observation, and post-staining wet-microscopic inspection is helpful for distinguishing white blood cells from infusions. The wet inspection is simple and quick to operate, but in clinical practice, the wet inspection is frequently carried out to miss fungus spores, clue cells and various bacteria.

The dry dyeing inspection of the vaginal secretion is to make a sample into a dry sheet and then observe the sample under a microscope, and because the sample is the dry sheet, an oil mirror which is 100 times of a microscope can be used for observation, and at the moment, various components are clear in form, particularly bacteria and fungi are very easy to distinguish, so that the dry dyeing inspection is the optimal scheme for analyzing the tangible components when the vaginal secretion is subjected to microecology inspection.

Among the dry staining of vaginal secretions, there are three methods of gram staining, jetty staining, and papanicolaou staining. Gram staining requires multiple steps of operations, which are time consuming, complex to operate, and complex to operate clinically. The Giemsa staining method is simple to operate, but the cell nuclei and cytoplasm after staining are blue and purple with cold tone, the color difference is small, the components in vaginal secretion are complex, and the similar tone needs to take longer time to identify each component.

In summary, the existing dry staining methods for vaginal secretions have poor staining results.

In the conventional preparation scheme of the Giemsa staining solution, the Giemsa staining solution and the Giemsa staining solution are required to be ground and dissolved by stirring with methanol, and then cured for 1 month for use. In the conventional preparation scheme, the curing time is long, the solubility of the dye in methanol is poor, and the dyeing effects of different batches of dyeing solutions are different, so that the application of the Jim-Sa-Rui-Shi dyeing solution to automatic film reading equipment is prevented.

Disclosure of Invention

The invention aims to provide a dry-method vaginal secretion dyeing liquid, a preparation method and a dyeing method thereof, and aims to solve the problem that the existing dry-method vaginal secretion dyeing method is poor in dyeing effect.

To achieve the above object, in a first aspect, the present invention provides a dry-method vaginal secretion staining solution comprising a first staining solution and a second staining solution;

The first dyeing liquid comprises blue dye, basic fuchsin, an organic solvent and deionized water;

the second staining solution comprises eosin, buffer solution, organic solvent and deionized water.

Wherein the first dyeing liquid comprises 1-3 parts of blue dye, 1-5 parts of basic fuchsin, 20-60 parts of organic solvent and 32-78 parts of deionized water.

Wherein the second staining solution comprises 1-5 parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water.

Wherein the blue dye comprises one or more of methylene blue, azure II, methyl green, methyl violet and toluidine blue;

the organic solvent comprises one or more of methanol, ethanol, glycol, glycerol, tetrahydrofuran and dimethyl sulfoxide.

Wherein the buffer solution comprises one or more of boric acid buffer solution, acetic acid buffer solution, phosphoric acid buffer solution, na2HPO 4-sodium citrate, citric acid-sodium citrate, potassium hydrogen phthalate-sodium hydroxide, potassium dihydrogen phosphate-sodium hydroxide, good's buffer solution, hank's buffer solution, amino acid buffer solution, TAPS buffer solution, bicine buffer solution, HEPS buffer solution, MES buffer solution and Tris buffer solution.

In a second aspect, the invention provides a method for preparing a dry-process vaginal secretion staining solution, comprising the following steps:

Weighing 1-3 parts of blue dye and 20-60 parts of organic solvent, stirring and dissolving for 3 hours at 20-30 ℃, adding 32-78 deionized water, uniformly mixing, and then weighing 1-5 parts of basic fuchsin, stirring and dissolving for 3 hours at 20-30 ℃ to obtain a first dyeing liquid;

1-5 parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain a second dyeing solution.

In a third aspect, the present invention provides a method of dry staining vaginal secretion staining solution comprising mixed staining and step staining.

Wherein the mixed staining comprises:

taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃;

after uniformly mixing the first staining solution and the second staining solution according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture to cover the dried sample, dyeing for 10-15S, pouring the staining solution, washing with purified water, drying, and observing under a microscope.

Wherein the step-wise staining comprises:

taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃;

Dripping 5-10 drops of the second staining solution to cover the dried sample, staining for 5-15S, pouring the staining solution, cleaning with purified water, dripping 5-10 drops of the first staining solution to cover the dried sample, staining for 5-15S, pouring the staining solution, cleaning with purified water, drying, and observing under a microscope.

The invention relates to a preparation method of a dry-method dyeing vaginal secretion dyeing liquid, which comprises a first dyeing liquid and a second dyeing liquid; the first dyeing liquid comprises blue dye, basic fuchsin, an organic solvent and deionized water; the second staining solution comprises eosin, buffer solution, organic solvent and deionized water.

The beneficial effects are that:

(1) Solves the problems of convenient operation and poor dyeing effect in the dry dyeing of vaginal secretion. The invention provides a preparation method of a dyeing liquid, which is convenient to operate and good in dyeing effect.

The invention uses 1 or more blue dyes and 2 red dyes, the red dyes contain eosin and basic fuchsin, and the eosin helps to strengthen the dyeing contrast of cell nuclei and cytoplasm when being dyed, and the basic fuchsin can enable the cytoplasm to be dyed into pink. The invention adjusts the proportion relation of the dye to make the cytoplasm become pink after staining the cell, the nucleus become blue-purple, the cytoplasm is slightly light in color and the nucleus, at this time, the cytoplasm and the nucleus have the difference of two dimensions of color tone temperature and color depth, so that the contrast ratio of the nucleus and the cytoplasm after staining is high, and the invention is convenient for the human eyes or instrument algorithm to capture and recognize.

(2) Solves the problems of good dyeing effect but poor storage stability of the dyeing liquid in the dyeing of vaginal secretion.

Basic fuchsin, blue dye and eosin can be reasonably matched under a specific pH environment, and can be used for dyeing out the difference of two dimensions of cytoplasm, cell nucleus color category and color depth. And because the dyed sample is a vaginal secretion sample, the pH of the vaginal secretion sample is acidic, so that the dyeing liquid system needs to be provided with a larger buffer capacity to ensure the normal coloring of each sample. However, when basic fuchsin is mixed with a weakly acidic to neutral buffer solution, significant discoloration of basic fuchsin will occur, and the greater the buffer capacity, the faster the rate at which basic fuchsin discoloration failure occurs. The combination of eosin and buffer solution does not influence the effect of strengthening nuclear dyeing, so the method for respectively preparing eosin and basic fuchsin into solution A and solution B can solve the contradictory relation between the dyeing effect and the stability of the dyeing solution.

(3) Solves the problems of good dyeing effect but batch-to-batch difference of dyeing liquid in the vaginal secretion dyeing.

The blue dye and the dyeing auxiliary eosin slowly react after being mixed to generate insoluble precipitate, and the solvent for dissolving the blue dye is a mixture of an organic solvent and deionized water. In order to ensure the solubility of the blue dye, a part of the organic solvent with volatility needs to be selected, but the solubility of eosin is sensitive to the proportion of the organic solvent, the proportion of the organic solvent is changed after the organic solvent volatilizes, eosin powder is separated out, and at the moment, two types of precipitates appear in the solution: and (3) precipitate and eosin powder precipitate generated after the blue dye reacts with eosin. When the blue dye and eosin are formulated, the reproducibility of the formulation of the solution is poor due to the difference in the volatilization degree of the organic solvent in the solvent at each formulation. The solvent is composed of the organic solvent which is not easy to volatilize and deionized water in the dyeing liquid B, and the proportion of the organic solvent is basically unchanged in the stirring and dissolving process, so that the blue dye and eosin are classified and prepared into the liquid A and the liquid B, and the problem of difference between solution preparation batches can be solved.

On the other hand, blue dyes have poor solubility in a single organic solvent, and basic fuchsin has general solubility in a single water or a single organic solvent. In order to achieve the effect of rapid dyeing, the drug concentrations of the blue dye and basic fuchsin need of preparing a high-concentration solution. In combination with these two points, the use of a single solvent is required to improve the efficiency of the solution A by increasing the dissolution temperature and prolonging the dissolution time. However, the organic solvent has potential safety hazard at high temperature, so that the problem of medicine dissolution of the solution A can be solved only by prolonging the dissolution time, and the preparation work efficiency of the solution A is very low. To increase the efficiency of formulation, it may be considered to increase the surfactant as a co-solvent or to change the solvent combination of the solution to improve drug dissolution. Because the dye designed by the patent is used for dry dyeing, the coloring effect can be poor due to the increase of the auxiliary dissolution of the surfactant, so the problem of the solubility of the solution A is improved by optimizing the solvent type of the solution A and the proportion between different solvents. The solvent of solution a is thus designed to consist of water and an organic solvent, wherein the organic solvent comprises one or more of methanol, ethanol, ethylene glycol, glycerol, tetrahydrofuran and dimethyl sulfoxide.

Drawings

In order to more clearly illustrate the embodiments of the invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic illustration of contrast enhancement of stained cells.

FIG. 2 is a graph showing the effect of mixing the first and second solutions according to the formulation.

FIG. 3 is a graph showing the effect of mixing the dye solutions C and D according to the composition of the medicines, but by exchanging the positions of basic fuchsin and eosin.

Fig. 4 is a schematic diagram of preparing a first dye solution, a second dye solution, and removing eosin to prepare a dye solution E and a dye solution F according to a formula.

Fig. 5 is a schematic diagram of preparing the first and second staining solutions according to the formulation and preparing the staining solutions G and H by removing basic fuchsin.

Fig. 6 is a schematic diagram of the first and second staining solutions, staining solution C and staining solution D.

Fig. 7 is a flow chart of a method for preparing a dry-method vaginal secretion staining solution provided by the invention.

Detailed Description

Embodiments of the present invention are described in detail below, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to like or similar elements or elements having like or similar functions throughout. The embodiments described below by referring to the drawings are illustrative and intended to explain the present invention and should not be construed as limiting the invention.

Please refer to fig. 1 to 7. FIG. 1 is a schematic illustration of contrast enhancement of stained cells. FIG. 2 is a graph showing the effect of mixing the first and second solutions according to the formulation. FIG. 3 is a graph showing the effect of mixing the dye solutions C and D according to the composition of the medicines, but by exchanging the positions of basic fuchsin and eosin. Fig. 4 is a schematic diagram of preparing a first dye solution, a second dye solution, and removing eosin to prepare a dye solution E and a dye solution F according to a formula. Fig. 5 is a schematic diagram of preparing the first and second staining solutions according to the formulation and preparing the staining solutions G and H by removing basic fuchsin. Fig. 6 is a schematic diagram of the first and second staining solutions, staining solution C and staining solution D. Fig. 7 is a flow chart of a method for preparing a dry-method vaginal secretion staining solution provided by the invention.

In a first aspect, the present invention provides a dry-method staining vaginal secretion staining solution comprising a first staining solution and a second staining solution; the first dyeing liquid comprises blue dye, basic fuchsin, an organic solvent and deionized water; the second staining solution comprises eosin, buffer solution, organic solvent and deionized water.

Specifically, the first dyeing liquid comprises 1-3 parts of blue dye, 1-5 parts of basic fuchsin, 20-60 parts of organic solvent and 32-78 parts of deionized water. The second staining solution comprises 1-5 parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water. The blue dye comprises one or more of methylene blue, azure II, methyl green, methyl violet and toluidine blue; the organic solvent comprises one or more of methanol, ethanol, glycol, glycerol, tetrahydrofuran and dimethyl sulfoxide. The buffer solution comprises one or more of boric acid buffer solution, acetic acid buffer solution, phosphoric acid buffer solution, na2HPO 4-sodium citrate, citric acid-sodium citrate, potassium hydrogen phthalate-sodium hydroxide, potassium dihydrogen phosphate-sodium hydroxide, good's buffer solution, hank's buffer solution, amino acid buffer solution, TAPS buffer solution, bicine buffer solution, HEPS buffer solution, MES buffer solution and Tris buffer solution.

In a second aspect, the invention provides a method for preparing a dry-process vaginal secretion staining solution, comprising the following steps:

S1, weighing 1-3 parts of blue dye and 20-60 parts of organic solvent, stirring and dissolving for 3 hours at 20-30 ℃, adding 32-78 deionized water, uniformly mixing, and then weighing 1-5 parts of basic fuchsin, stirring and dissolving for 3 hours at 20-30 ℃ to obtain a first dyeing liquid;

S2, 1-5 parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain a second dyeing solution.

In a third aspect, the present invention provides a method of dry staining vaginal secretion staining solution comprising mixed staining and step staining.

The mixed staining includes:

s101, taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃;

S102, uniformly mixing the first staining solution and the second staining solution according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture to cover the dried sample, dyeing for 10-15S, pouring the staining solution, washing with purified water, drying, and observing under a microscope.

The step-wise staining comprises:

S201, taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃;

S202, dripping 5-10 drops of the second staining solution to cover the dried sample, staining for 5-15S, pouring the staining solution, cleaning with purified water, dripping 5-10 drops of the first staining solution to cover the dried sample, staining for 5-15S, pouring the staining solution, cleaning with purified water, drying, and observing under a microscope.

The beneficial effects are that:

(1) Solves the problems of convenient operation and poor dyeing effect in the dry dyeing of vaginal secretion. The invention provides a preparation method of a dyeing liquid, which is convenient to operate and good in dyeing effect.

The invention uses 1 or more blue dyes and 2 red dyes, the red dyes contain eosin and basic fuchsin, and the eosin helps to strengthen the dyeing contrast of cell nuclei and cytoplasm when being dyed, and the basic fuchsin can enable the cytoplasm to be dyed into pink. The invention adjusts the proportion relation of the dye to make the cytoplasm become pink after staining the cell, the nucleus become blue-purple, the cytoplasm is slightly light in color and the nucleus, at this time, the cytoplasm and the nucleus have the difference of two dimensions of color tone temperature and color depth, so that the contrast ratio of the nucleus and the cytoplasm after staining is high, and the invention is convenient for the human eyes or instrument algorithm to capture and recognize.

(2) Solves the problems of good dyeing effect but poor storage stability of the dyeing liquid in the dyeing of vaginal secretion.

Basic fuchsin, blue dye and eosin can be reasonably matched under a specific pH environment, and can be used for dyeing out the difference of two dimensions of cytoplasm, cell nucleus color category and color depth. And because the dyed sample is a vaginal secretion sample, the pH of the vaginal secretion sample is acidic, so that the dyeing liquid system needs to be provided with a larger buffer capacity to ensure the normal coloring of each sample. However, when basic fuchsin is mixed with a weakly acidic to neutral buffer solution, significant discoloration of basic fuchsin will occur, and the greater the buffer capacity, the faster the rate at which basic fuchsin discoloration failure occurs. The combination of eosin and buffer solution does not influence the effect of strengthening nuclear dyeing, so the method for respectively preparing eosin and basic fuchsin into solution A and solution B can solve the contradictory relation between the dyeing effect and the stability of the dyeing solution.

(3) Solves the problems of good dyeing effect but batch-to-batch difference of dyeing liquid in the vaginal secretion dyeing.

The blue dye and the dyeing auxiliary eosin slowly react after being mixed to generate insoluble precipitate, and the solvent for dissolving the blue dye is a mixture of an organic solvent and deionized water. In order to ensure the solubility of the blue dye, a part of the organic solvent with volatility needs to be selected, but the solubility of eosin is sensitive to the proportion of the organic solvent, the proportion of the organic solvent is changed after the organic solvent volatilizes, eosin powder is separated out, and at the moment, two types of precipitates appear in the solution: and (3) precipitate and eosin powder precipitate generated after the blue dye reacts with eosin. When the blue dye and eosin are formulated, the reproducibility of the formulation of the solution is poor due to the difference in the volatilization degree of the organic solvent in the solvent at each formulation. The solvent is composed of the organic solvent which is not easy to volatilize and deionized water in the dyeing liquid B, and the proportion of the organic solvent is basically unchanged in the stirring and dissolving process, so that the blue dye and eosin are classified and prepared into the liquid A and the liquid B, and the problem of difference between solution preparation batches can be solved.

Experimental results:

the contrast of the stained cells was enhanced as shown in fig. 1.

Balancing the stability and dyeing effect of the dyeing liquid:

examples:

2.1 preparing a first staining solution, a second staining solution according to a scheme and staining a sample:

According to parts by weight, 1-3 parts of blue dye, 1-5 parts of basic fuchsin, 20-60 parts of organic solvent and 32-78 parts of deionized water are respectively weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain a first dyeing liquid.

1-5 Parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are respectively weighed according to parts by weight, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain a second dyeing solution. The buffer capacity of the second staining solution is 0.05 mol/L-0.08 mol/L, and the pH value is 5.0-7.5.

The dyeing experiment is carried out by adopting a mixed dyeing mode:

Taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃. After uniformly mixing the first staining solution and the second staining solution according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture to cover the dried sample, dyeing for 10-15S, pouring the staining solution, washing with purified water, drying, and observing under a microscope.

3 Cases of clinical samples are prepared, each case is sampled and dried into 2 dry slices, and the dry slices of the samples are placed at 15-25 ℃ for sealing and storage. And before dying the dried sample, drying the sample at 40-60 ℃ for 10-15S, and then carrying out a dying experiment.

1 Sample was dyed immediately after the completion of the preparation of the staining solution, and another 1 sample was dyed 30 days after the preparation of the staining solution, and the dyeing effect was as shown in fig. 2.

2.2 Exchange basic fuchsin and eosin in solutions a, B according to the drug composition based on the protocol:

According to parts by weight, 1-3 parts of blue dye, 1-5 parts of eosin, 20-60 parts of organic solvent and 32-78 parts of deionized water are respectively weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid C.

1-5 Parts of basic fuchsin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are respectively weighed according to parts by weight, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid D. The buffer capacity of the dyeing liquid D is 0.05mol/L to 0.08mol/L, and the pH value is 5.0 to 7.5.

The dyeing experiment is carried out by adopting a mixed dyeing mode:

Taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃. After evenly mixing the dyeing liquid C and the dyeing liquid D according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture to cover the dried sample, dyeing for 10-15S, pouring the dyeing liquid, washing with purified water, drying and observing under a microscope.

3 Cases of clinical samples are prepared, each case is sampled and dried into 2 dry slices, and the dry slices of the samples are placed at 15-25 ℃ for sealing and storage. And before dying the dried sample, drying the sample at 40-60 ℃ for 10-15S, and then carrying out a dying experiment.

1 Sample was dyed immediately after the completion of the preparation of the staining solution, and another 1 sample was dyed 30 days after the preparation of the staining solution, and the dyeing effect was as shown in fig. 3. After 30 days of standing basic fuchsin in combination with the buffer solution, basic fuchsin was significantly discolored.

2.3 In the invention, the main function of the eosin component is to enhance the dyeing contrast of cell nucleus and cytoplasm, and the eosin is removed and then is dyed by preparing the dyeing liquid, and the dyeing effect is compared:

According to parts by weight, 1-3 parts of blue dye, 1-5 parts of basic fuchsin, 20-60 parts of organic solvent and 32-78 parts of deionized water are respectively weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid E.

1-5 Parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are respectively weighed according to parts by weight, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid F. The buffer capacity of the dyeing liquid F is 0.05mol/L to 0.08mol/L, and the pH value is 5.0 to 7.5.

The dyeing experiment is carried out by adopting a mixed dyeing mode:

Taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃. After evenly mixing the dyeing liquid E and the dyeing liquid F according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture to cover the dried sample, dyeing for 10-15S, pouring the dyeing liquid, washing with purified water, drying and observing under a microscope.

And (3) taking the first dyeing liquid and the second dyeing liquid to dye in the same mode, and comparing the dyeing effect after eosin removal. As shown in fig. 4.

As can be seen from a comparison of the same samples, the cytoplasm of both experimental groups was pink and the nucleus blue, but the samples with eosin deleted had poor contrast of the nucleus cytoplasm. Eosin thus has a co-dyeing effect in the first dyeing liquor, dyeing B of the present invention.

2.4 In the present invention, the cytoplasm was pink using basic fuchsin as the main cytoplasmic staining agent, and the comparative example was performed in the group 2 in which basic fuchsin was not added and the staining solution of the present invention:

according to the weight portions, 1 to 3 portions of blue dye, 20 to 60 portions of organic solvent and 32 to 78 portions of deionized water are respectively weighed and evenly mixed at the temperature of 20 to 30 ℃ and stirred and dissolved for 3 hours to obtain dyeing liquid G.

1-5 Parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are respectively weighed according to parts by weight, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid H. The buffer capacity of the dyeing liquid F is 0.05mol/L to 0.08mol/L, and the pH value is 5.0 to 7.5.

The dyeing experiment is carried out by adopting a mixed dyeing mode:

Taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃. Mixing the dyeing solution I and the dyeing solution J according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture after uniform mixing, covering and drying the sample, dyeing for 10-15S, pouring the dyeing solution, cleaning with purified water, drying and observing under a microscope.

Basic fuchsin can make the cell value obviously pink, and improve the cell number and cell contrast. As shown in fig. 5.

2.5 In the present invention, after separation of eosin from blue dye, reproducibility of dye solution preparation was improved, and the following is a test example:

According to parts by weight, 1-3 parts of blue dye and 20-60 parts of organic solvent are respectively weighed, stirred and dissolved for 3 hours at 20-30 ℃, deionized water 32-78 is added and uniformly mixed, and then 1-5 parts of basic fuchsin are weighed, stirred and dissolved for 3 hours at 20-30 ℃ to obtain a first dyeing liquid.

1-5 Parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are respectively weighed according to parts by weight, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain a second dyeing solution. The buffer capacity of the second staining solution is 0.05 mol/L-0.08 mol/L, and the pH value is 5.0-7.5.

According to parts by weight, 1-3 parts of blue dye, 1-5 parts of eosin, 20-60 parts of organic solvent and 32-78 parts of deionized water are respectively weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid C.

1-5 Parts of basic fuchsin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are respectively weighed according to parts by weight, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain dyeing liquid D. The buffer capacity of the dyeing liquid D is 0.05mol/L to 0.08mol/L, and the pH value is 5.0 to 7.5.

And (3) preparing the solution A, the solution B, the solution C and the solution D for 1 time respectively at the same room temperature according to the same preparation process, mixing and dyeing the same sample by taking the solution A and the solution B as a group and the solution C and the solution D as a group after the dyeing dark night after the preparation is finished. The staining results are shown in fig. 6.

After the eosin is separated from the blue dye according to the method, the preparation repeatability of the solution is improved.

The above disclosure is only illustrative of a dry method for dyeing vaginal secretion, and the preparation method and dyeing method thereof, but is not limited thereto, and those skilled in the art will appreciate that all or part of the procedures for implementing the above embodiments are equivalent and still fall within the scope of the invention.

Claims (9)

1. A dry-method vaginal secretion staining solution is characterized in that,

Comprises a first staining solution and a second staining solution;

The first dyeing liquid comprises blue dye, basic fuchsin, an organic solvent and deionized water;

the second staining solution comprises eosin, buffer solution, organic solvent and deionized water.

2. The dry-process vaginal secretion dye according to claim 1, wherein,

The first dyeing liquid comprises 1-3 parts of blue dye, 1-5 parts of basic fuchsin, 20-60 parts of organic solvent and 32-78 parts of deionized water.

3. The dry-process vaginal secretion dye according to claim 2, wherein,

The second staining solution comprises 1-5 parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water.

4. The dry-process vaginal secretion dye according to claim 3, wherein,

The blue dye comprises one or more of methylene blue, azure II, methyl green, methyl violet and toluidine blue;

the organic solvent comprises one or more of methanol, ethanol, glycol, glycerol, tetrahydrofuran and dimethyl sulfoxide.

5. The dry-process vaginal secretion dye according to claim 4, wherein,

The buffer solution comprises one or more of boric acid buffer solution, acetic acid buffer solution, phosphoric acid buffer solution, na2HPO 4-sodium citrate, citric acid-sodium citrate, potassium hydrogen phthalate-sodium hydroxide, potassium dihydrogen phosphate-sodium hydroxide, good's buffer solution, hank's buffer solution, amino acid buffer solution, TAPS buffer solution, bicine buffer solution, HEPS buffer solution, MES buffer solution and Tris buffer solution.

6. A method for preparing a dry-dyed vaginal secretion dye, according to claim 5, comprising the steps of:

Weighing 1-3 parts of blue dye and 20-60 parts of organic solvent, stirring and dissolving for 3 hours at 20-30 ℃, adding 32-78 deionized water, uniformly mixing, and then weighing 1-5 parts of basic fuchsin, stirring and dissolving for 3 hours at 20-30 ℃ to obtain a first dyeing liquid;

1-5 parts of eosin, 1-5 parts of buffer solution, 10-20 parts of organic solvent and 70-88 parts of deionized water are weighed, uniformly mixed at 20-30 ℃, and stirred and dissolved for 3 hours to obtain a second dyeing solution.

7. A method for dry-dyeing vaginal discharge staining solution, comprising the step of dry-dyeing the vaginal discharge staining solution according to claim 5,

Including mixed staining and step staining.

8. The method for dry-dyeing vaginal secretion dye according to claim 7, wherein,

The mixed staining includes:

taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃;

after uniformly mixing the first staining solution and the second staining solution according to the proportion of 1:1-1.5, dripping 5-10 drops of the mixture to cover the dried sample, dyeing for 10-15S, pouring the staining solution, washing with purified water, drying, and observing under a microscope.

9. The method for dry-dyeing vaginal secretion dye according to claim 7, wherein,

The step-wise staining comprises:

taking a vaginal secretion sample, adding 1-3 mL of physiological saline for eluting, taking 1-2 drops of the eluted solution on a glass slide, and drying the sample at the temperature of 45-60 ℃;

Dripping 5-10 drops of the second staining solution to cover the dried sample, staining for 5-15S, pouring the staining solution, cleaning with purified water, dripping 5-10 drops of the first staining solution to cover the dried sample, staining for 5-15S, pouring the staining solution, cleaning with purified water, drying, and observing under a microscope.