A staining solution for exfoliated cells and its preparation method and application

Abstract

The present invention discloses a staining solution for exfoliated cells, a preparation method thereof and an application thereof, wherein the staining solution comprises the following components in parts by weight: 10 to 21 parts of a reducing agent, 0.03 to 0.05 parts of folic acid, 0.03 to 0.06 parts of methylene blue, 1 to 3 parts of a solvent, 1.5 to 3 parts of a base, and 75 to 90 parts of water; the pH of the staining solution is 8 to 14. The staining solution in the present invention is an alkaline solution, which has the advantages of not being easily oxidized, good staining effect, long reagent storage time, high detection sensitivity and specificity, etc. In addition, the methylene blue in the staining solution of the present invention has good solubility, solves the problem of methylene blue being difficult to dissolve in the prior art, and can be widely used in special cytological staining of exfoliated cells, such as coarse screening inspection of clinical routine lesions such as cervix, vagina, and vulva.

Description

Technical Field

[0001] The invention relates to the biological field, and in particular to a staining solution for exfoliated cells and a preparation method and application thereof. Background Art

[0002] Folate receptors are less expressed or not expressed in normal cells, but highly expressed on the surface of some diseased cells. Folate receptor-mediated special staining is a new and more convenient detection method that has developed rapidly in recent years. In this method, the main components of the staining solution are folic acid derivatives, reduced methylene blue, etc. Due to the mediation of folic acid receptors, methylene blue has a high affinity for tumor cells and melanin, and its redox property makes methylene blue show different color changes in the redox state of tumor tissue. The reaction principle is: if there is a high degree of folic acid expression on the surface of epithelial tissue exfoliated cells, the folic acid in the staining solution will quickly enter the cell cytoplasm through the folic acid receptors on the cell surface, and the reduced methylene blue will also enter the cell cytoplasm. Folic acid is reduced to tetrahydrofolate and participates in the metabolism of one-carbon units. Tumor cells are in a strong oxidative stress environment, and the reduced methylene blue is oxidized by the reactive oxygen species (ROS) widely present in the cells to become blue oxidized methylene blue. Due to the entry of many biological molecules, the osmotic pressure in the cells increases, causing the oxidized methylene blue to escape from the cells, which can appear blue on the staining solution. The appearance of blue staining indicates that the expression of folate receptors on the surface of the sample cells is increased, the reactive oxygen pressure in the cells is increased, and the ferritin content is high, indicating that the exfoliated cells may have abnormal lesions and need further testing.

[0003] Based on the principle of this color change method, many related special staining solutions are currently available on the market. The special staining solutions available on the market are all acidic solutions. It is generally recognized in the art that under acidic conditions, the staining solution is more likely to bind to the folate receptors on the surface of cancer cells, thereby entering the cancer cells; and under the oxidative stress environment of tumor cells, the reduced colorless methylene blue will be converted into a blue or blue-black oxidized state, thereby achieving the diagnosis of cancer cells. The methylene blue aqueous solution is an alkaline solution. In the prior art, an acidic solution is added when preparing the staining solution. + Acid-base neutralization reaction will occur at high concentrations. When the dyeing liquid mixture is acidic and air is present, the dyeing liquid will be oxidized faster, resulting in poor dyeing effect and short reagent storage time. Therefore, the acidic dyeing liquid in the prior art has the following disadvantages: (1) the dyeing liquid is easily oxidized and discolored during storage, thereby affecting the accuracy of the test results; (2) the dyeing liquid is unstable, has a short storage period, and has poor storage stability; (3) the dyeing effect of the dyeing liquid is not ideal, and the dyeing results are inaccurate. Summary of the invention

[0004] In order to overcome the problem of adding acidic solution to the staining solution in the prior art, one of the purposes of the present invention is to provide a staining solution for exfoliated cells. The staining solution in the present invention is alkaline, and the methylene blue (oxidized state) and reducing agent (reducing sugar and ascorbic acid, etc.) in the staining solution are converted into methylene white (reduced state) under alkaline conditions. The staining solution is not easily oxidized, has a good staining effect, and the reagent has a long shelf life and is more stable.

[0005] A second object of the present invention is to provide a method for preparing a staining solution for exfoliated cells.

[0006] A third object of the present invention is to provide a tumor cell detection kit.

[0007] A fourth object of the present invention is to provide a method for using a staining solution for exfoliated cells.

[0008] A fifth object of the present invention is to provide a staining solution for exfoliated cells for use in tumor cell detection materials.

[0009] In order to achieve the above object, the technical solution adopted by the present invention is:

[0010] The first aspect of the present invention is to provide a staining solution for exfoliated cells, the staining solution comprising the following components in parts by weight: 10 to 21 parts of a reducing agent, 0.03 to 0.05 parts of folic acid, 0.03 to 0.06 parts of methylene blue, 1 to 3 parts of a solvent, 1.5 to 3 parts of an alkali, and 75 to 90 parts of water; the pH of the staining solution is 8 to 14.

[0011] Preferably, the weight proportion of the reducing agent is 10 to 20; further preferably, the weight proportion of the reducing agent is 10 to 18; still further preferably, the weight proportion of the reducing agent is 10 to 16.

[0012] Preferably, the weight portion of folic acid is 0.03 to 0.045 parts.

[0013] Preferably, the amount of the solvent used is 1 to 2 parts.

[0014] Preferably, the amount of water used is 80 to 90 parts; more preferably, the amount of water used is 80 to 88 parts.

[0015] Preferably, the pH of the dyeing solution is 10-13; further preferably, the pH of the dyeing solution is 11.75-13.

[0016] Preferably, the base is selected from at least one of sodium hydroxide, potassium hydroxide, calcium hydroxide and barium hydroxide; more preferably, the base is sodium hydroxide.

[0017] Preferably, the reducing agent is selected from at least one of monosaccharides and ascorbic acid; further preferably, the reducing agent includes monosaccharides and ascorbic acid.

[0018] Preferably, the weight ratio of the monosaccharide to ascorbic acid is (50-75):1.

[0019] Preferably, the monosaccharide is selected from at least one of fructose, glucose, galactose and ribose.

[0020] Preferably, the solvent includes dimethyl sulfoxide. The present invention is an alkaline system, and the solubility of methylene blue under alkaline conditions is very poor. The present invention uses dimethyl sulfoxide as a solvent to achieve that methylene blue still has good solubility under alkaline conditions.

[0021] Preferably, the exfoliated cells include cervical exfoliated cells, vaginal exfoliated cells, and vulvar exfoliated cells.

[0022] The second aspect of the present invention is to provide a method for preparing the staining solution for exfoliated cells provided in the first aspect of the present invention, comprising the following steps:

[0023] The reducing agent, folic acid, solvent and water are mixed, and then mixed with methylene blue and alkali to prepare the dyeing solution.

[0024] Preferably, the preparation method comprises the following steps:

[0025] S1: Weigh the reducing agent, folic acid, methylene blue and solvent respectively;

[0026] S2: After dissolving the reducing agent and folic acid in part of the water, adding the solvent and continuing to mix;

[0027] S3: adding methylene blue to the mixed solution of step S2, and then mixing;

[0028] S4: add alkali to step S3 and stir to mix evenly, add remaining water and mix.

[0029] Preferably, the amount of the partial water added in step S2 accounts for more than 80% of the total amount of water; further preferably, the amount of the partial water added in step S2 accounts for more than 90% of the total amount of water.

[0030] Preferably, the base is selected from at least one of sodium hydroxide, potassium hydroxide, calcium hydroxide and barium hydroxide; more preferably, the base is sodium hydroxide.

[0031] The third aspect of the present invention is to provide a tumor cell detection kit, comprising the staining solution for exfoliated cells provided in the first aspect of the present invention.

[0032] The fourth aspect of the present invention is to provide a method for using the staining solution for exfoliated cells provided in the first aspect of the present invention, wherein a detection material containing the staining solution is mixed with the exfoliated cells, and the color change time and color of the detection material are recorded.

[0033] The fifth aspect of the present invention is to provide the use of the staining solution for exfoliated cells provided in the first aspect of the present invention in tumor cell detection materials.

[0034] Preferably, the tumor cells include epithelial tissue tumor cells.

[0035] The beneficial effects of the present invention are as follows: the staining solution in the present invention is an alkaline solution, which has the advantages of not being easily oxidized, good staining effect, long reagent storage time, high detection sensitivity and specificity, etc. In addition, the methylene blue in the staining solution of the present invention has good solubility, which solves the problem of methylene blue being difficult to dissolve in the prior art, and can be widely used in special cytological staining of exfoliated cells, such as coarse screening examination of clinical routine lesions such as the cervix, vagina, and vulva.

[0036] In addition, the present invention makes the dyeing solution alkaline for the first time, achieving an unexpected effect, promoting the reaction of methylene blue with a reducing agent (reducing sugar, ascorbic acid, etc.) under alkaline conditions to generate methylene white, thereby improving the stability of the dyeing solution. Furthermore, under alkaline conditions, the dyeing solution accelerates the binding of folic acid to the receptor, resulting in a faster dyeing rate, better dyeing effect, and greatly improved dyeing accuracy.

[0037] The preparation method of the dyeing solution of the present invention is simple, easy to operate, has mild reaction conditions, does not require special reaction equipment, and is suitable for industrial mass production. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 This is a picture of the exfoliated cells dipped in the staining solution in Example 3 and reacted for 0 seconds;

[0039] Figure 2 This is a color image of exfoliated cells dipped in the staining solution in Example 3 and reacted for 35 seconds;

[0040] Figure 3 This is a picture of the reaction of 0s with the staining solution in Comparative Example 1 to dip the exfoliated cells;

[0041] Figure 4 This is a color image of exfoliated cells dipped in the staining solution in Comparative Example 1 and reacted for 65 seconds. DETAILED DESCRIPTION

[0042] The specific implementation of the present invention is further described in detail below in conjunction with the accompanying drawings and examples, but the implementation and protection of the present invention are not limited thereto. It should be noted that if there are processes that are not particularly described in detail below, they can be implemented or understood by those skilled in the art with reference to the prior art. If the manufacturer of the reagents or instruments used is not indicated, they are deemed to be conventional products that can be purchased commercially.

[0043] Example 1

[0044] The staining solution for exfoliated cells in this example is made of the following components in weight percentage: 10% monosaccharide, 0.2% ascorbic acid, 0.03% folic acid, 0.03% methylene blue, 1% dimethyl sulfoxide, 1.5% sodium hydroxide, and 87.24% purified water; wherein the monosaccharide is a mixture of glucose and fructose in a weight ratio of 1:1, and the pH value of the staining solution in this example is 11.75.

[0045] The staining solution for exfoliated cells in this example is prepared by the following preparation method, which specifically includes the following steps:

[0046] (1) According to a preset weight percentage, the monosaccharide is stirred with a portion of water until it is completely dissolved and then mixed;

[0047] (2) adding ascorbic acid to the solution obtained in step (1) according to a preset weight percentage, and stirring until it is completely dissolved and mixed;

[0048] (3) adding folic acid to the solution obtained in step (2) according to a preset weight percentage, and stirring until completely dissolved and mixed;

[0049] (4) adding dimethyl sulfoxide to the solution obtained in step (3) according to a preset weight percentage, and stirring to mix;

[0050] (5) adding methylene blue to the solution obtained in step (4) according to a preset weight percentage, and stirring for 5-10 min until it is completely dissolved and mixed;

[0051] (6) Sodium hydroxide was added to the solution obtained in step (5) and stirred until completely dissolved and mixed, and the remaining water was added to prepare the staining solution for exfoliated cells in this example. The pH value of the staining solution was measured by a pH meter to be 11.75.

[0052] Example 2

[0053] The staining solution for exfoliated cells in this example is made of the following components in weight percentage: 15% monosaccharide, 0.2% ascorbic acid, 0.03% folic acid, 0.03% methylene blue, 1% dimethyl sulfoxide, 2% sodium hydroxide, and 81.74% purified water; wherein the monosaccharide is a mixture of glucose and fructose in a weight ratio of 2:1, and the pH value of the staining solution in this example is 12.25.

[0054] The staining solution for exfoliated cells in this example is prepared by the following preparation method, which specifically includes the following steps:

[0055] (1) According to a preset weight percentage, the monosaccharide is stirred with a portion of water until it is completely dissolved and then mixed;

[0056] (2) adding ascorbic acid to the solution obtained in step (1) according to a preset weight percentage, and stirring until it is completely dissolved and mixed;

[0057] (3) adding folic acid to the solution obtained in step (2) according to a preset weight percentage, and stirring until completely dissolved and mixed;

[0058] (4) adding dimethyl sulfoxide to the solution obtained in step (3) according to a preset weight percentage, and stirring to mix;

[0059] (5) adding methylene blue to the solution obtained in step (4) according to a preset weight percentage, and stirring for 5-10 min until it is completely dissolved and mixed;

[0060] (6) Sodium hydroxide was added to the solution obtained in step (5) and stirred until completely dissolved and mixed, and the remaining water was added to prepare the staining solution for exfoliated cells in this example. The pH value of the staining solution was measured by a pH meter to be 12.25.

[0061] Example 3

[0062] The staining solution for exfoliated cells in this example is made of the following components in weight percentage: 15% monosaccharide, 0.3% ascorbic acid, 0.045% folic acid, 0.06% methylene blue, 1% dimethyl sulfoxide, 2.25% sodium hydroxide, and 81.345% purified water; wherein the monosaccharide is a mixture of glucose and fructose in a weight ratio of 2:1, and the pH value of the staining solution in this example is 12.50.

[0063] The staining solution for exfoliated cells in this example is prepared by the following preparation method, which specifically includes the following steps:

[0064] (1) According to a preset weight percentage, the monosaccharide is stirred with a portion of water until it is completely dissolved and then mixed;

[0065] (2) adding ascorbic acid to the solution obtained in step (1) according to a preset weight percentage, and stirring until it is completely dissolved and mixed;

[0066] (3) adding folic acid to the solution obtained in step (2) according to a preset weight percentage, and stirring until completely dissolved and mixed;

[0067] (4) adding dimethyl sulfoxide to the solution obtained in step (3) according to a preset weight percentage, and stirring to mix;

[0068] (5) adding methylene blue to the solution obtained in step (4) according to a preset weight percentage, and stirring for 5-10 min until it is completely dissolved and mixed;

[0069] (6) Sodium hydroxide was added to the solution obtained in step (5) and stirred until completely dissolved and mixed, and the remaining water was added to obtain the staining solution for exfoliated cells in this example. The pH value of the staining solution was measured by a pH meter to be 12.50.

[0070] Example 4

[0071] The staining solution for exfoliated cells in this example is made of the following components in weight percentage: 15% monosaccharide, 0.2% ascorbic acid, 0.045% folic acid, 0.06% methylene blue, 1% dimethyl sulfoxide, 2.5% sodium hydroxide, and 81.195% purified water; wherein the monosaccharide is a mixture of glucose and fructose in a weight ratio of 2:1, and the pH value of the staining solution in this example is 12.75.

[0072] The staining solution for exfoliated cells in this example is prepared by the following preparation method, which specifically includes the following steps:

[0073] (1) According to a preset weight percentage, the monosaccharide is stirred with a portion of water until it is completely dissolved and then mixed;

[0074] (2) adding ascorbic acid to the solution obtained in step (1) according to a preset weight percentage, and stirring until it is completely dissolved and mixed;

[0075] (3) adding folic acid to the solution obtained in step (2) according to a preset weight percentage, and stirring until completely dissolved and mixed;

[0076] (4) adding dimethyl sulfoxide to the solution obtained in step (3) according to a preset weight percentage, and stirring to mix;

[0077] (5) adding methylene blue to the solution obtained in step (4) according to a preset weight percentage, and stirring for 5-10 min until it is completely dissolved and mixed;

[0078] (6) Sodium hydroxide was added to the solution obtained in step (5) and stirred until completely dissolved and mixed, and the remaining water was added to obtain the staining solution for exfoliated cells in this example. The pH value of the staining solution was measured by a pH meter to be 12.75.

[0079] Example 5

[0080] The staining solution for exfoliated cells in this example is made of the following components in weight percentage: 15% monosaccharide, 0.2% ascorbic acid, 0.045% folic acid, 0.06% methylene blue, 1% dimethyl sulfoxide, 3% sodium hydroxide, and 80.695% purified water; wherein the monosaccharide is a mixture of glucose and fructose in a weight ratio of 2:1, and the pH value of the staining solution in this example is 13.00.

[0081] The staining solution for exfoliated cells in this example is prepared by the following preparation method, which specifically includes the following steps:

[0082] (1) According to a preset weight percentage, the monosaccharide is stirred with a portion of water until it is completely dissolved and then mixed;

[0083] (2) adding ascorbic acid to the solution obtained in step (1) according to a preset weight percentage, and stirring until it is completely dissolved and mixed;

[0084] (3) adding folic acid to the solution obtained in step (2) according to a preset weight percentage, and stirring until completely dissolved and mixed;

[0085] (4) adding dimethyl sulfoxide to the solution obtained in step (3) according to a preset weight percentage, and stirring to mix;

[0086] (5) adding methylene blue to the solution obtained in step (4) according to a preset weight percentage, and stirring for 5-10 min until it is completely dissolved and mixed;

[0087] (6) Sodium hydroxide was added to the solution obtained in step (5) and stirred until completely dissolved and mixed, and the remaining water was added to prepare the staining solution for exfoliated cells in this example. The pH value of the staining solution was measured by a pH meter and was 13.00.

[0088] Comparative Example 1

[0089] The staining solution for exfoliated cells in this example is made of the following components in weight percentage: 15% monosaccharide, 0.2% ascorbic acid, 0.045% folic acid, 0.06% methylene blue, 3% dimethyl sulfoxide, 0.5% hydrochloric acid, and 81.195% purified water; wherein the monosaccharide is a mixture of glucose and fructose in a weight ratio of 2:1, and the pH value of the staining solution in this example is 6.00.

[0090] The staining solution for exfoliated cells in this example is prepared by the following preparation method, which specifically includes the following steps:

[0091] (1) According to a preset weight percentage, the monosaccharide is stirred with a portion of water until it is completely dissolved and then mixed;

[0092] (2) adding ascorbic acid to the solution obtained in step (1) according to a preset weight percentage, and stirring until it is completely dissolved and mixed;

[0093] (3) adding folic acid to the solution obtained in step (2) according to a preset weight percentage, and stirring until completely dissolved and mixed;

[0094] (4) adding dimethyl sulfoxide to the solution obtained in step (3) according to a preset weight percentage, and stirring to mix;

[0095] (5) adding methylene blue to the solution obtained in step (4) according to a preset weight percentage, and stirring for 5-10 min until it is completely dissolved and mixed;

[0096] (6) 0.5% by weight of 3 mol / L hydrochloric acid was added to the solution obtained in step (5), and the remaining water was added to prepare the staining solution for exfoliated cells in this example. The pH value of the staining solution was measured by a pH meter and was 6.00.

[0097] Performance Test:

[0098] (1) Dyeing efficiency test

[0099] The dyeing solutions prepared in Examples 1-5 and Comparative Example 1 were respectively taken, and the dyeing efficiency of each dyeing solution was tested. The specific testing method is as follows:

[0100] First, use a cotton swab to dip into the staining liquid in Examples 1-5 and Comparative Example 1 respectively, and then dip into the exfoliated cells, and observe and record the color change of the cotton swab. This is different from the detection method in the prior art. The detection method in the prior art is: it is necessary to dip into the staining liquid first, apply the staining liquid to the position to be tested, and then take a sample. In this method, the staining liquid is in contact with the air for a long time, and the air will affect the stability and accuracy of the test results. In the detection method of the present invention, the staining liquid is in contact with the air for a shorter time, the accuracy of the test results is higher, and the staining liquid will not directly contact the human body, and the detection process is safer. Among them, the test method for staining efficiency is: take 4 portions of each staining liquid, dip the staining liquid with a cotton swab, let it stand in the air, use a stopwatch to record the color change time of the staining liquid, and take the average value. The specific test results are shown in Figures 1 to 4 ,in, Figure 1 This is a picture of the exfoliated cells dipped in the staining solution in Example 3 and reacted for 0 seconds; Figure 2 This is a color image of exfoliated cells dipped in the staining solution in Example 3 and reacted for 35 seconds; Figure 3 This is a picture of the reaction of 0s with the staining solution in Comparative Example 1 to dip the exfoliated cells; Figure 4 The color development diagram is a 65s reaction of exfoliated cells dipped in the staining solution in Comparative Example 1. The results measured according to the above test method are shown in Table 1.

[0101] Table 1 Dyeing efficiency test results of Examples 1-5 and Comparative Example 1

[0102] staining solution Average color change time How to use Example 1 42.3s Dip in staining solution before sampling Example 2 41.7s Dip in staining solution before sampling Example 3 40.2s Dip in staining solution before sampling Example 4 40.3s Dip in staining solution before sampling Example 5 40.6s Dip in staining solution before sampling Comparative Example 1 61.8s Dip in staining solution before sampling

[0103] As can be seen from Table 1, the average color change time of the dyeing solution prepared in Examples 1-5 is 40.2-42.3s, while the average color change time of the dyeing solution prepared in Comparative Example 1 under the same test conditions is 61.8s. Therefore, compared with the acidic dyeing solution in the prior art (i.e., the dyeing solution in Comparative Example 1), the alkaline dyeing solution in the present invention has higher dyeing efficiency.

[0104] (2) Antioxidant capacity test

[0105] The dyeing solutions prepared in Examples 1-5 and Comparative Example 1 were stored in a 42°C water bath, and the antioxidant properties of each sample were determined by iodine titration. The iodine titration method is specifically as follows: the iodine titration method is divided into indirect iodine titration and direct iodine titration. We used the direct iodine titration method for testing. The direct iodine titration method uses iodine element titration: C6H8O6+I2=C6H6O6+2HI. The specific test results are shown in Table 2 below.

[0106] Table 2 Antioxidant capacity test results of the dyeing solution in Examples 1-5 and Comparative Example 1

[0107]

[0108] As can be seen from Table 2, after 3 days, the antioxidant capacity of the dyeing solutions in Examples 1 to 5 and Comparative Example 1 decreased, among which the antioxidant capacity of the dyeing solution prepared in Comparative Example 1 decreased by more than 60% within 3 days; while the antioxidant capacity of the dyeing solutions prepared in Examples 1 to 5 decreased by less than 10% after storage for 40 days, indicating that the antioxidant capacity of the dyeing solution of the alkaline system in the present invention is significantly stronger than that of the dyeing solution of the acidic system in the prior art, with good stability and long storage time.

[0109] (3) Sensitivity and specificity test

[0110] Sensitivity and specificity testing refers to staining tests on sections of patients with confirmed early epithelial tissue tumor cell lesions. The measured staining success rate is the sensitivity and specificity test result. The sensitivity test is used to illustrate the staining of epithelial tissue tumor cells by the staining solution. The higher the sensitivity, the stronger the ability to detect abnormal cells. The specificity test is used to illustrate the accuracy of the staining solution. The higher the specificity, the higher the consistency rate of the stained abnormal cells with the pathological test results. The sensitivity and specificity calculation formula is:

[0111] Sensitivity = number of true positives / (number of true positives + number of false positives) × 100%;

[0112] Specificity = number of true negatives / (number of true negatives + number of false positives) × 100%;

[0113] 48 sections of patients with confirmed epithelial cell tissue tumor cell lesions and 112 sections of patients with non-epithelial cell tissue tumor cell lesions were selected for staining test, TCT and HPV examination. The operation steps of the staining experiment are: first use a cotton swab to dip into the staining solution in Example 5 and Comparative Example 1, and then dip into the exfoliated cells, and then observe the color change of the cotton swab, and then test the sensitivity and specificity of the staining solution. Under the same conditions, the existing TCT and HPV technologies were used for testing, and the specific test results are shown in Table 3 below:

[0114] Table 3 Sensitivity and specificity test results of Example 5 and Comparative Example 1

[0115] Inspection method Sensitivity Specificity Example 5 82.1％ 90.1％ Comparative Example 1 66.4％ 86.2％ TCT 66.2％ 97.9％ HPV 96.5％ 63.6％

[0116] It can be seen from Table 3 that the staining sensitivity and specificity of the staining solution prepared in the present invention are higher than those of the acidic system staining solution in Comparative Example 1, and compared with TCT and HPV testing technologies, the staining solution of the present invention does not require instruments and equipment during testing, and the product cost is low and the patient compliance is higher.

[0117] The above is a detailed description of the embodiments of the present invention, but the present invention is not limited to the above embodiments. Various changes can be made within the knowledge of ordinary technicians in the relevant technical field without departing from the purpose of the present invention. In addition, the embodiments of the present invention and the features in the embodiments can be combined with each other without conflict.

Claims

1. A staining solution for exfoliated cells, characterized in that: The dyeing solution comprises the following components in parts by weight: 10 to 21 parts of a reducing agent, 0.03 to 0.05 parts of folic acid, 0.03 to 0.06 parts of methylene blue, 1 to 3 parts of a solvent, 1.5 to 3 parts of a base, and 75 to 90 parts of water; the pH of the dyeing solution is 8 to 14; the reducing agent is selected from at least one of a monosaccharide and ascorbic acid; The solvent includes dimethyl sulfoxide; The alkali is selected from at least one of sodium hydroxide, potassium hydroxide, calcium hydroxide and barium hydroxide.

2. The staining solution for exfoliated cells according to claim 1, characterized in that: The pH of the dyeing solution is 10-13.

3. The staining solution for exfoliated cells according to claim 1, characterized in that: The monosaccharide is selected from at least one of fructose, glucose, galactose and ribose.

4. The method for preparing a staining solution for exfoliated cells according to any one of claims 1 to 3, characterized in that: The following steps are involved: The reducing agent, folic acid, solvent and water are mixed, and then mixed with methylene blue and alkali to prepare the dyeing solution.

5. A tumor cell detection kit, characterized in that: The invention comprises the staining solution for exfoliated cells as described in any one of claims 1 to 3.

6. The method for using the staining solution for exfoliated cells according to any one of claims 1 to 3, characterized in that: The detection material containing the dye solution is mixed with the exfoliated cells, and the color change time and color of the detection material are recorded.

7. Use of the staining solution for exfoliated cells according to any one of claims 1 to 3 in tumor cell detection materials.

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