Eosin staining solution formula

Abstract

The invention discloses a formula of an eosin staining solution, and every 1000 ml of the eosin staining solution comprises the following components: 5-8 g of eosin which is any one of water-soluble eosin Y or alcohol-soluble eosin Y; 2.5 to 3.5 g of potassium dichromate; 5-25 g of picric acid; 5 to 60 ml of absolute ethyl alcohol; the balance is water, and the water is any one of deionized water or distilled water. According to the eosin staining solution formula disclosed by the invention, the absolute ethyl alcohol not only can promote good dissolution of all the components and effectively ensure a good uniform mixing effect among all the components, but also can be used as a dehydrating agent and can play a role in dehydrating and fixing cells, so that a cell environment in tissues is similar to the tissues, cell coloring in the tissues is facilitated, and the quality of the cells in the tissues is improved. The dyeing liquid has a certain level dyeing effect, is clear in coloring, avoids overdyeing, uneven dyeing and other conditions, utilizes the synergistic effect of potassium dichromate and picric acid, promotes the dissolution of each component, ensures the stability of the dyeing liquid, is simpler to prepare, is not easy to fade, and has a longer period of validity.

Description

Technical Field

[0001] The invention relates to the technical field of dyeing liquid, in particular to a formula of eosin dyeing liquid. Background Art

[0002] Eosin staining solution can dissociate into negatively charged anions in water, which combine with the positive charge of the amino group of the protein, thereby dyeing the cytoplasm into different shades of red. At present, the eosin staining solutions used are mainly: aqueous solution of water-soluble eosin, 95% ethanol solution of alcohol-soluble eosin, and 80% ethanol solution of water-soluble eosin and phloxine. In the US patent document US2011053279A1, a staining method for detecting proteins in polyacrylamide gel and its organic stain are also disclosed, wherein the disclosed staining agent is one of eosin Y or phloxine B. However, all of the above-mentioned staining solutions using eosin as a staining agent have the following problems: (1) The composition of the dye is single, resulting in unclear layers when staining the cytoplasm and extracellular matrix; (2) Due to the low staining intensity of eosin, acetic acid needs to be added to the above-mentioned eosin staining solution as an accelerator to enhance the staining intensity. However, the addition of the accelerator will weaken the staining color layers, further exacerbating the problem of unclear staining layers; at the same time, after the addition of the accelerator, not only does the acidic substance need to be washed off during the subsequent processing of the slices, but the acidic substance will also cause the slices to fade easily during the storage process when it remains, and the obtained slices cannot be stored for a long time.

[0003] Therefore, the existing hematoxylin and eosin staining solution is prone to produce metal film or dye precipitation, resulting in uneven staining and false blue staining. At the same time, the staining level of the eosin staining solution is not clear and the coloring effect is not good, which leads to blurred background of the stained tissue sections or cell smears, making it difficult to read the film. Therefore, we propose a formula of eosin staining solution. Summary of the invention

[0004] The main purpose of the present invention is to provide a formula of eosin staining solution, which can effectively solve the problems in the background technology.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0007] Eosin 5-8g;

[0008] Potassium dichromate 2.5-3.5g;

[0009] Picric acid 5-25g;

[0010] Anhydrous ethanol 5-60ml;

[0011] The balance is water.

[0012] Preferably, each 1000 ml of the eosin staining solution includes the following components:

[0013] Eosin 8g;

[0014] Potassium dichromate 3.2g;

[0015] Picric acid 18g;

[0016] 45ml of anhydrous ethanol;

[0017] The balance is water.

[0018] Preferably, the water is any one of deionized water or distilled water.

[0019] Preferably, the eosin is any one of water-soluble eosin Y or alcohol-soluble eosin Y.

[0020] Preferably, the method further comprises the following preparation steps:

[0021] S1: Take eosin, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio;

[0022] S2: adding eosin to anhydrous ethanol solution and performing water bath sonication to obtain eosin alcohol solution;

[0023] S3: Add eosin alcohol solution to potassium dichromate solution, add picric acid while stirring, filter after water bath ultrasonication, and add water to obtain eosin staining solution.

[0024] Preferably, in S2, the water bath temperature of the anhydrous ethanol solution in which eosin is added is 30° C., and the water bath ultrasonic time is 2 min.

[0025] Preferably, in S3, the water bath temperature for adding the eosin alcohol solution to the potassium dichromate solution is 30° C., the water bath temperature for adding picric acid is 35-40° C., and the water bath ultrasonic time is 3 min.

[0026] Compared with the prior art, the present invention has the following beneficial effects:

[0027] 1. The present invention adds anhydrous ethanol, which not only promotes the good dissolution of each component, effectively ensures the good mixing effect between the components, but also can be used as a dehydrating agent to dehydrate and fix the cells, so that the cell environment in the tissue is similar to the tissue, which is beneficial to the coloring of the cells in the tissue, and makes the eosin staining solution have a certain leveling effect, clear coloring, and avoids over-staining, uneven staining and the like.

[0028] 2. The eosin staining solution reagent of the present invention is simple to prepare. The components are added step by step at different temperatures, which not only ensures the homogeneity of each solution, but also prevents the occurrence of caking and the like. At the same time, the ultrasonic method is used to increase the mixing between the components and the solubility of the components.

[0029] 3. The present invention reduces the types of solutions, making the preparation of the dyeing solution simpler, and utilizes the synergistic effect of potassium dichromate and picric acid to promote the dissolution of each component, thereby ensuring the stability of the dyeing solution, making it less prone to fading, and having a longer shelf life. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 The present invention is a flow chart of the preparation of an eosin staining solution formula. DETAILED DESCRIPTION

[0031] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further explained below in conjunction with specific implementation methods.

[0032] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0033] 5-8 g of eosin, wherein the eosin is any one of water-soluble eosin Y and alcohol-soluble eosin Y;

[0034] Potassium dichromate 2.5-3.5g;

[0035] Picric acid 5-25g;

[0036] Anhydrous ethanol 5-60ml;

[0037] The balance is water, and the water is any one of deionized water or distilled water.

[0038] like Figure 1 As shown, the following preparation steps are also included:

[0039] S1: Take eosin, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio;

[0040] S2: Add eosin to anhydrous ethanol solution, set the water bath temperature at 30°C, and perform water bath ultrasonication for 2 min to obtain eosin alcohol solution;

[0041] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add water to obtain the eosin staining solution.

[0042] Example 1

[0043] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0044] Water-soluble eosin Y 8g;

[0045] Potassium dichromate 3.2g;

[0046] Picric acid 18g;

[0047] 45ml of anhydrous ethanol;

[0048] The balance was deionized water.

[0049] like Figure 1 As shown, the following preparation steps are also included:

[0050] S1: Take water-soluble eosin Y, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio;

[0051] S2: adding water-soluble eosin Y into anhydrous ethanol solution, setting the water bath temperature at 30°C and the water bath ultrasonic time for 2 min to obtain eosin alcohol solution;

[0052] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add deionized water to obtain the eosin staining solution.

[0053] The staining solution of this embodiment takes only 2 minutes to apply. The tissue section staining experiment proves that the background is clear, the layers are distinct, the staining effect is good, the slides are easy to read, and the slides are not easy to fade, so they are convenient for long-term storage.

[0054] Example 2

[0055] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0056] Alcohol-soluble eosin Y 8g;

[0057] Potassium dichromate 3.2g;

[0058] Picric acid 18g;

[0059] 45ml of anhydrous ethanol;

[0060] The balance is distilled water.

[0061] like Figure 1 As shown, the following preparation steps are also included:

[0062] S1: Take alcohol-soluble eosin Y, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio;

[0063] S2: Add alcohol-soluble eosin Y into anhydrous ethanol solution, set the water bath temperature at 30°C, and perform water bath ultrasonication for 2 min to obtain eosin alcohol solution;

[0064] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add distilled water to obtain the eosin staining solution.

[0065] The staining solution of this embodiment takes only 3 minutes to apply. The tissue section staining experiment proves that the background is clear, the layers are distinct, the staining effect is good, the slides are easy to read, and the slides are not easy to fade, so they are convenient for long-term storage.

[0066] Example 3

[0067] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0068] Alcohol-soluble eosin Y 8g;

[0069] Potassium dichromate 3.5g;

[0070] Picric acid 25g;

[0071] 60ml of anhydrous ethanol;

[0072] The balance was deionized water.

[0073] The following preparation steps are also included:

[0074] S1: Take alcohol-soluble eosin Y, potassium dichromate, picric acid, anhydrous ethanol and deionized water in a selected ratio;

[0075] S2: Add alcohol-soluble eosin Y into anhydrous ethanol solution, set the water bath temperature at 30°C, and perform water bath ultrasonication for 2 min to obtain eosin alcohol solution;

[0076] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add deionized water to obtain the eosin staining solution.

[0077] The staining solution of this embodiment takes only 5 minutes to apply. The tissue section staining experiment proves that the background is clear, the layers are distinct, the staining effect is good, the slides are easy to read, and the slides are not easy to fade, so they are convenient for long-term storage.

[0078] Example 4

[0079] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0080] Water-soluble eosin Y 8g;

[0081] Potassium dichromate 3.5g;

[0082] Picric acid 25g;

[0083] 60ml of anhydrous ethanol;

[0084] The balance was distilled water.

[0085] The following preparation steps are also included:

[0086] S1: Take water-soluble eosin Y, potassium dichromate, picric acid, anhydrous ethanol and deionized water in a selected ratio;

[0087] S2: adding water-soluble eosin Y into anhydrous ethanol solution, setting the water bath temperature at 30°C and the water bath ultrasonic time for 2 min to obtain eosin alcohol solution;

[0088] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add distilled water to obtain the eosin staining solution.

[0089] The staining solution of this embodiment takes only 4 minutes to apply. The tissue section staining experiment proves that the background is clear, the layers are distinct, the staining effect is good, the slides are easy to read, and the slides are not easy to fade, so they are convenient for long-term storage.

[0090] Example 5

[0091] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0092] Water-soluble eosin Y 5g;

[0093] Potassium dichromate 2.5g;

[0094] Picric acid 5g;

[0095] 5 ml of anhydrous ethanol;

[0096] The balance is deionized water.

[0097] The following preparation steps are also included:

[0098] S1: Take water-soluble eosin Y, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio;

[0099] S2: adding water-soluble eosin Y into anhydrous ethanol solution, setting the water bath temperature at 30°C and the water bath ultrasonic time for 2 min to obtain eosin alcohol solution;

[0100] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add ionized water to obtain the eosin staining solution.

[0101] The time required for applying the staining solution of this embodiment is only 6 minutes. The tissue section staining experiment proves that the background is clear, the layers are distinct, the staining effect is good, the slices are easy to read, and the staining is not easy to fade, so it is convenient for long-term storage.

[0102] Example 6

[0103] A formula of eosin staining solution, wherein each 1000 ml of the eosin staining solution comprises the following components:

[0104] 5 g of alcohol-soluble eosin Y, wherein the eosin is any one of water-soluble eosin Y or eosin Y;

[0105] Potassium dichromate 2.5g;

[0106] Picric acid 5g;

[0107] 5 ml of anhydrous ethanol;

[0108] The balance is distilled water.

[0109] The following preparation steps are also included:

[0110] S1: Take alcohol-soluble eosin Y, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio;

[0111] S2: Add alcohol-soluble eosin Y into anhydrous ethanol solution, set the water bath temperature at 30°C, and perform water bath ultrasonication for 2 min to obtain eosin alcohol solution;

[0112] S3: When the water bath temperature is 30°C, add the eosin alcohol solution to the potassium dichromate solution, add picric acid while stirring, the water bath temperature is 35-40°C, the water bath ultrasonic time is 3 minutes, filter, and add distilled water to obtain the eosin staining solution.

[0113] The time required for applying the staining solution of this embodiment is only 6 minutes. The tissue section staining experiment proves that the background is clear, the layers are distinct, the staining effect is good, the slices are easy to read, and the staining is not easy to fade, so it is convenient for long-term storage.

[0114] The invention adds anhydrous ethanol, which can not only promote the good dissolution of each component and effectively ensure the good mixing effect between each component, but also can be used as a dehydrating agent to dehydrate and fix cells, so that the cell environment in the tissue is similar to the tissue, which is beneficial to the coloring of cells in the tissue, and the eosin staining solution has a certain leveling effect, clear coloring, and avoids the occurrence of over-dyeing, uneven dyeing and the like; the eosin staining solution reagent is simple to prepare, and the components are added step by step at different temperatures, which not only ensures the homogeneity of each solution, but also prevents the occurrence of agglomeration and the like, and simultaneously adopts an ultrasonic method to increase the mixing between each component and the solubility of the component; the types of solutions are reduced, so that the preparation of the dyeing solution is simpler, and the synergistic effect of potassium dichromate and picric acid is utilized to promote the dissolution of each component, ensure the stability of the dyeing solution, and the dyeing solution is not easy to fade and has a long validity period.

[0115] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It should be understood by those skilled in the art that the present invention is not limited to the above embodiments. The above embodiments and descriptions are only for explaining the principles of the present invention. Without departing from the spirit and scope of the present invention, the present invention may have various changes and improvements, which fall within the scope of the present invention to be protected. The scope of protection of the present invention is defined by the attached claims and their equivalents.

Claims

1. A formula of eosin staining solution, characterized in that: Each 1000 ml of the eosin staining solution includes the following components: Eosin 5-8g; Potassium dichromate 2.5-3.5g; Picric acid 5-25g; Anhydrous ethanol 5-60ml; The balance is water.

2. A eosin staining solution formulation according to claim 1, characterized in that: Each 1000 ml of the eosin staining solution includes the following components: Eosin 8g; Potassium dichromate 3.2g; Picric acid 18g; 45ml of anhydrous ethanol; The balance is water.

3. A eosin staining solution formulation according to claim 1, characterized in that: The water is any one of deionized water or distilled water.

4. A eosin staining solution formulation according to claim 1, characterized in that: The eosin is any one of water-soluble eosin Y and alcohol-soluble eosin Y.

5. A eosin staining solution formulation according to claim 1, characterized in that: The following preparation steps are also included: S1: Take eosin, potassium dichromate, picric acid, anhydrous ethanol and water in a selected ratio; S2: adding eosin to anhydrous ethanol solution and performing water bath sonication to obtain eosin alcohol solution; S3: Add eosin alcohol solution to potassium dichromate solution, add picric acid while stirring, filter after water bath ultrasonication, and add water to obtain eosin staining solution.

6. A eosin staining solution formulation according to claim 5, characterized in that: In the S2, the water bath temperature of the anhydrous ethanol solution to which eosin is added is 30° C., and the water bath ultrasonic time is 2 min.

7. A eosin staining solution formulation according to claim 5, characterized in that: In the S3, the water bath temperature for adding the eosin alcohol solution to the potassium dichromate solution is 30° C., the water bath temperature for adding picric acid is 35-40° C., and the water bath ultrasonic time is 3 min.

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