CN114624084A - Preparation and staining method of pathological tissue section staining kit - Google Patents
Preparation and staining method of pathological tissue section staining kit Download PDFInfo
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Images
Classifications
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
- G01N2001/302—Stain compositions
Abstract
The invention relates to the technical field of pathological tissue section staining, and discloses a preparation method of a pathological tissue section staining kit, wherein the staining kit comprises filtered stable liquid, hematoxylin staining solution, differentiation solution, bluing solution and eosin staining solution; the stabilizing solution comprises 2-20 parts of organic acid, 0-300 parts of polyhydric alcohol and 1000 parts of water, and the pH value of the stabilizing solution is adjusted to 2.5-3.5 by sodium hydroxide solution; the hematoxylin staining solution comprises 2-10 parts of hematoxylin, 300 parts of polyhydric alcohol 150-. According to the preparation method of the pathological tissue section staining kit, hematoxylin staining solution does not have a metal film or precipitate in the using process by adjusting the proportion of the mordant in the components, so that the workload of technical personnel is greatly reduced, the staining efficiency of cell nucleuses is improved, the kit can be used for a long time, and meanwhile, chromatin of the cell nucleuses is stained meticulously, so that the staining effect of the cell nucleuses is improved.
Description
Technical Field
The invention relates to the technical field of pathological tissue section staining, in particular to a preparation and staining method of a pathological tissue section staining kit.
Background
Hematoxylin-eosin staining (HE staining) is one of the staining methods commonly used in paraffin section technology. The hematoxylin staining solution is alkaline, and mainly makes the chromatin in the cell nucleus and the nucleic acid in the cytoplasm bluish; eosin is an acid dye that primarily reddens components in the cytoplasm and extracellular matrix. The HE staining method is the most basic and widely used technical method in histology, embryology, pathology teaching and scientific research. The operation steps in the hematoxylin-eosin dyeing method mainly comprise slicing, baking, dewaxing, rehydration, stabilizing solution, hematoxylin dyeing, washing, differentiation, washing, blueing returning, washing, eosin dyeing, dehydration, transparence and sealing.
In the conventional HE dyeing operation, the slices are dewaxed and hydrated and then directly enter the hematoxylin staining solution to perform cell nucleus staining, on one hand, the last step of the hydration is to pass through flowing tap water containing hypochlorous acid with strong oxidizing property and accumulate continuously, so that the oxidation of hematoxylin in the hematoxylin staining solution is accelerated to form hematoxylin, thereby shortening the service life of the hematoxylin staining solution and obviously increasing the use amount of the hematoxylin staining solution, and on the other hand, when the sliced tissues enter a complex staining solution environment from a relatively simple tap water solution environment, the staining effects of different tissues have certain difference, even the condition of easy decolorization occurs, and the staining effect of cell nuclei is seriously reduced, so that the problems need to be improved.
The current hematoxylin staining solution commonly has the following problems:
1. a metal film is easy to generate on the surface of the hematoxylin staining solution, or a dye is precipitated, so that stains are easy to exist on the surface of the slice after hematoxylin staining, the stain is difficult to remove, and the phenomena of uneven staining and slice reading are caused;
2. for a metal film generated by hematoxylin staining solution, the metal film needs to be filtered and removed, so that the workload of technicians is increased, and time and reagents are wasted;
3. the phenomenon of nucleoplasm co-dyeing is easy to occur, and the dyeing effect is not good;
4. the dyeing stability is poor, the color is easy to fade, and the use requirement of long-term storage cannot be met;
5. different requirements are required for dyeing time in different seasons and temperatures, and the clinical application can be started only by searching for proper dyeing time, so that the use effect is seriously reduced. Although the existing eosin dye formula is simple to operate, the dyeing time is long, the effective period of dye liquor is short, glacial acetic acid is required to be added for use, the dosage of the glacial acetic acid is not easy to master, the addition amount is small and cannot achieve the effect of promoting dyeing, the phenomena of irregular color and easy fading of section dyeing are caused when the glacial acetic acid is added too much, and the dyeing effect cannot be stored for a long time.
Disclosure of Invention
Technical problem to be solved
Aiming at the defects of the prior art, the invention provides a preparation and staining method of a pathological tissue section staining kit, which is used for solving the problems.
(II) technical scheme
In order to solve the technical problems, the invention provides the following technical scheme: a preparation method of a pathological tissue section staining kit comprises a filtered stabilizing solution, a hematoxylin staining solution, a differentiation solution, a bluing solution and an eosin staining solution;
the stabilizing solution comprises 2-20 parts of organic acid, 0-300 parts of polyhydric alcohol and 1000 parts of 700-water, and the pH value of the stabilizing solution is adjusted to 2.5-3.5 by sodium hydroxide solution;
the hematoxylin dyeing solution comprises 2-10 parts of hematoxylin, 300 parts of polyhydric alcohol 150-;
the differentiation solution comprises 3-70 parts of acid, 0-700 parts of polyalcohol and 300-1000 parts of water, and the pH value of the differentiation solution is adjusted to 1.5-3.5 by a sodium hydroxide solution;
the blue returning liquid comprises 0.5-20 parts of alkaline compound, 0-300 parts of polyalcohol and 1000 parts of 700-sodium bicarbonate water, and the pH value of the blue returning liquid is 1.5-11.0;
the eosin dye solution comprises 1-10 parts of eosin Y, 0-950 parts of ethanol, 0-4 parts of glacial acetic acid and 50-1000 parts of water, and the pH value of the eosin dye solution is 2.5-5.0.
Preferably, in the stabilizing solution, the organic acid includes glacial acetic acid, tartaric acid and citric acid, and the polyhydric alcohol includes ethanol, ethylene glycol and propylene glycol.
Preferably, in the hematoxylin staining solution, the polyhydric alcohol comprises ethanol, ethylene glycol, propylene glycol and glycerol, the oxidizing agent comprises sodium iodate and potassium iodate, the mordant comprises aluminum sulfate, potassium aluminum sulfate and ammonium aluminum sulfate, and the organic acid comprises glacial acetic acid and citric acid.
Preferably, in the differentiation solution, the acid includes hydrochloric acid, tartaric acid and glacial acetic acid, and the polyol includes ethanol, ethylene glycol and propylene glycol.
Preferably, in the bluing liquid, the basic compounds include ammonia water, sodium bicarbonate, lithium carbonate, magnesium sulfate, and tris (hydroxymethyl) aminomethane, and the polyhydric alcohols include ethylene glycol and propylene glycol.
Preferably, in the eosin dye solution, the eosin Y comprises alcohol-soluble eosin and water-soluble eosin.
A staining method of a pathological tissue section staining kit comprises staining by using the staining kit.
(III) advantageous effects
Compared with the prior art, the invention provides a preparation and staining method of a pathological tissue section staining kit, which has the following beneficial effects:
1. according to the preparation method of the pathological tissue section staining kit, hematoxylin staining solution does not generate a metal film in the use process by adjusting the proportion of the mordant in the components, so that the workload of technical personnel is greatly reduced, the staining efficiency of cell nucleuses is improved, the kit can be used for a long time, the staining of the chromatin of the cell nucleuses is delicate, and the staining effect of the cell nucleuses is improved.
2. According to the preparation method of the pathological tissue section staining kit, the reagents of all components are optimally combined and proportioned, so that the hematoxylin staining solution has stronger selectivity to cell nucleuses, the tissue staining structure is clear, the staining is durable, and the long-time preservation requirement can be met through an aging test (60 ℃).
3. The preparation method of the pathological tissue section staining kit has the advantages that under the condition of the same reagent, the temperature is low, the longer the time for achieving the same staining effect is, the temperature can influence the staining speed, the sensitivity of the hematoxylin staining solution to the temperature is greatly reduced, the staining time does not need to be adjusted due to the temperature change, and the using effect of the hematoxylin staining solution is improved.
4. The preparation method of the pathological tissue section staining kit is not easy to form crystalline substances in the staining process of automatic equipment through the filtering and the antioxidation of the stabilizing solution,
5. the staining method of the pathological tissue section staining kit is convenient to operate, short in staining time, capable of completing the whole staining process within 30 +/-5 min, and suitable for large-scale popularization and application in full-automatic equipment.
Drawings
FIG. 1 is a schematic view showing that no metal film is generated when the hematoxylin solution is placed for one month in the preparation method of the pathological tissue section staining kit provided by the invention;
FIG. 2 is a view showing the staining effect of a tissue slice in the preparation method of the staining kit for a pathological tissue slice according to the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 100mL/L of ethanol, 15mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.8 by using a sodium hydroxide solution;
hematoxylin staining solution, which specifically comprises the following components: 150mL/L of ethanol, 3.0g/L of hematoxylin, 30.0g/L of aluminum potassium sulfate, 0.3g/L of sodium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 2.3 by glacial acetic acid;
the differentiation liquid specifically comprises: 4.2mL/L of hydrochloric acid and the balance, and adjusting the pH value to 1.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 2g/L of sodium bicarbonate and the balance of water, wherein the pH value of the solution is 8.5;
the eosin dye solution specifically comprises the following components: 950mL/L of absolute ethyl alcohol, 4.0g/L of alcohol-soluble eosin Y and the balance of water, wherein the pH value of the solution is 3.0.
Example 2
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 125mL/L of ethanol, 2.5g/L of tartaric acid and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
hematoxylin staining solution, which specifically comprises the following components: 200mL/L of ethylene glycol, 5.0g/L of hematoxylin, 44.0g/L of aluminum sulfate, 0.5g/L of sodium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 2.2 by glacial acetic acid;
the differentiation liquid specifically comprises: 50mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 0.5g/L of lithium carbonate and the balance of water, wherein the pH value of the solution is 10.5;
the eosin dye solution specifically comprises the following components: 950mL/L of absolute ethyl alcohol, 3.5g/L of alcohol-soluble eosin Y and the balance of water, and the pH value of the solution is 3.05.
Dip dyeing method for paraffin section
Example 3
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 75mL/L of ethanol, 75mL/L of glycol, 3.5g/L of citric acid and the balance of water, and adjusting the pH to 3.0 by using a sodium hydroxide solution;
hematoxylin staining solution, which comprises the following specific steps: 125mL/L of ethanol, 125mL/L of ethylene glycol, 7.0g/L of hematoxylin, 70.0g/L of aluminum ammonium sulfate, 0.7g/L of potassium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 2.4 by citric acid;
the differentiation liquid specifically comprises: tartaric acid 15.0g/L and the balance water, and adjusting the pH to 2.3 by sodium hydroxide solution;
the blue returning liquid specifically comprises: 4.5mL/L of ammonia water and the balance of water, wherein the pH value of the solution is 10.8;
the eosin dye solution specifically comprises the following components: 950mL/L of absolute ethyl alcohol, 3.0g/L of alcohol-soluble eosin Y, 0.1mL of glacial acetic acid and the balance of water, wherein the pH value of the solution is 3.1.
Example 4
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 200mL/L of propylene glycol, 20mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
hematoxylin staining solution, which specifically comprises the following components: 200mL/L of propylene glycol, 9.0g/L of hematoxylin, 90.0g/L of aluminum sulfate, 0.9g/L of potassium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to be 2.1 by glacial acetic acid;
the differentiation liquid specifically comprises: 50mL/L of glacial acetic acid, 700mL/L of ethanol and the balance of water, and adjusting the pH to 2.8 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 2.0g/L of sodium bicarbonate, 10.0g/L of magnesium sulfate and the balance of water, wherein the pH value of the solution is 7.8;
the eosin dye solution specifically comprises the following components: water-soluble eosin Y10g/L, 1.0mL glacial acetic acid and balance water, the solution pH was 4.8.
Example 5
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 100mL/L of ethanol, 15mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.8 by using a sodium hydroxide solution;
hematoxylin staining solution, which specifically comprises the following components: 250mL/L of ethylene glycol, 2.0g/L of hematoxylin, 20.0g/L of aluminum potassium sulfate, 0.2g/L of potassium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 2.2 by glacial acetic acid;
the differentiation liquid specifically comprises: 50mL/L of glacial acetic acid, 500mL/L of ethanol and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 15g/L of trihydroxymethyl aminomethane and the balance of water, and adjusting the pH value to 9.0 by a hydrochloric acid solution;
the eosin dye solution specifically comprises the following components: 950mL/L of absolute ethyl alcohol, 2.0g/L of alcohol-soluble eosin Y and the balance of water, and the pH value of the solution is 3.0.
Example 6
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 125mL/L of ethylene glycol, 2.5g/L of tartaric acid and the balance of water, and adjusting the pH value to 2.5 by using a sodium hydroxide solution;
hematoxylin staining solution is environment-friendly and mercury-free, and specifically comprises the following components: 250mL/L of ethylene glycol, 4.0g/L of hematoxylin, 20.0g/L of aluminum potassium sulfate, 20.0g/L of aluminum ammonium sulfate, 0.4g/L of potassium iodate and the balance of water, and the pH is adjusted to 2.2 by glacial acetic acid;
the differentiation liquid specifically comprises: 50mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 1.5g/L of sodium bicarbonate, 15.0g/L of magnesium sulfate and the balance of water, wherein the pH value of the solution is 7.9;
the eosin dye solution specifically comprises the following components: 850mL/L of absolute ethyl alcohol, 2.5g/L of alcohol-soluble eosin Y and the balance of water, wherein the pH value of the solution is 3.05.
Example 7
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 75mL/L of ethanol, 75mL/L of glycol, 3.5g/L of citric acid and the balance of water, and adjusting the pH to 3.0 by using a sodium hydroxide solution;
hematoxylin staining solution is environment-friendly and mercury-free, and specifically comprises the following components: 125mL/L of ethanol, 125mL/L of ethylene glycol, 6.0g/L of hematoxylin, 50.0g/L of aluminum sulfate, 0.6g/L of sodium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 2.2 by glacial acetic acid;
the differentiation liquid specifically comprises: tartaric acid 15.0g/L and the balance water, and adjusting the pH to 2.3 by sodium hydroxide solution;
the blue returning liquid specifically comprises: 1.0g/L of lithium carbonate and the balance of water, wherein the pH value of the solution is 10.8;
the eosin dye solution specifically comprises the following components: 850mL/L of absolute ethyl alcohol, 3.0g/L of alcohol-soluble eosin Y and the balance of water, wherein the pH value of the solution is 3.10.
Example 8
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 200mL/L of propylene glycol, 20mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
hematoxylin staining solution is environment-friendly and mercury-free, and specifically comprises the following components: 150mL/L of ethanol, 8.0g/L of hematoxylin, 80.0g/L of aluminum ammonium sulfate, 0.8g/L of sodium iodate, 150mL/L of glycerol and the balance of water, and adjusting the pH to 2.2 by glacial acetic acid;
the differentiation liquid specifically comprises: 4.2mL/L of hydrochloric acid and the balance of water, and adjusting the pH value to 1.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 5.0mL/L of ammonia water and the balance of water, wherein the pH value of the solution is 10.8;
the eosin dye solution specifically comprises the following components: 850mL/L of water ethanol, 3.5g/L of alcohol-soluble eosin Y and the balance of water, wherein the pH value of the solution is 3.15.
Comparative example 1
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 125mL/L of ethylene glycol, 2.5g/L of tartaric acid and the balance of water, and adjusting the pH value to 2.5 by using a sodium hydroxide solution;
hematoxylin staining solution, which specifically comprises the following components: 150mL/L of ethanol, 3.0g/L of hematoxylin, 30.0g/L of aluminum potassium sulfate, 0.3g/L of sodium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 1.5 by glacial acetic acid;
the differentiation liquid specifically comprises: 4.2mL/L of hydrochloric acid and the balance of water, and adjusting the pH value to 1.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 2g/L of sodium bicarbonate and the balance of water, wherein the pH value of the solution is 8.5;
the eosin dye solution specifically comprises the following components: 950mL/L of absolute ethyl alcohol, 5.0g/L of alcohol-soluble eosin Y and the balance of water, wherein the pH value of the solution is 3.0.
Comparative example 1 compared with example 1, when the pH of hematoxylin in comparative example 1 is 1.5, the cell nucleus is difficult to color; in comparative example 1, alcohol-soluble eosin Y is 5.0g/L, more precipitates are generated, and in example 1, the tissue staining structure is clear and the staining is durable.
Comparative example 2
A preparation method of a pathological tissue section staining kit comprises the steps of filtering a stabilizing solution 1L, a hematoxylin staining solution 1L, a differentiation solution 1L, a bluing solution 1L and an eosin staining solution 1L in the staining kit;
the stabilizing solution specifically comprises: 125mL/L of glycol, 2.5g/L of tartaric acid and the balance of water, and adjusting the pH value to 2.5 by using a sodium hydroxide solution;
hematoxylin staining solution, which specifically comprises the following components: 200mL/L of ethylene glycol, 5.0g/L of hematoxylin, 44.0g/L of aluminum sulfate, 0.5g/L of sodium iodate, 50mL/L of glycerol and the balance of water, and adjusting the pH to 3.5 by sodium hydroxide;
the differentiation liquid specifically comprises: 50mL/L of glacial acetic acid and the balance of water, and adjusting the pH to 2.5 by using a sodium hydroxide solution;
the blue returning liquid specifically comprises: 0.5g/L of lithium carbonate and the balance of water, wherein the pH value of the solution is 10.5;
the eosin dye solution specifically comprises the following components: 950mL/L of absolute ethyl alcohol, 3.5g/L of alcohol-soluble eosin Y and the balance of water, wherein the pH value of the solution is 3.05.
Comparative example 2 in comparison with example 2, when the pH of hematoxylin in comparative example 2 was 3.5, the cell nuclei were hardly colored.
Example 9
The staining method of the pathological tissue section staining kit comprises the step of staining by using the staining kit, is convenient to operate and short in staining time, can be completed in 30 +/-5 min in the whole staining process, and is suitable for large-scale popularization and application in full-automatic equipment.
It is to be noted that the term "comprises," "comprising," or any other variation thereof is intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. A preparation method of a pathological tissue section staining kit is characterized by comprising the following steps: the staining reagent box comprises filtered stable liquid, hematoxylin staining solution, differentiation solution, blue returning solution and eosin staining solution;
the stabilizing solution comprises 2-20 parts of organic acid, 0-300 parts of polyhydric alcohol and 1000 parts of 700-water, and the pH value of the stabilizing solution is adjusted to 2.5-3.5 by sodium hydroxide solution;
the hematoxylin dyeing solution comprises 2-10 parts of hematoxylin, 300 parts of polyhydric alcohol 150-;
the differentiation solution comprises 3-70 parts of acid, 0-700 parts of polyalcohol and 300-1000 parts of water, and the pH value of the differentiation solution is adjusted to 1.5-3.5 by a sodium hydroxide solution;
the blue returning liquid comprises 0.5-20 parts of alkaline compound, 0-300 parts of polyalcohol and 1000 parts of 700-sodium bicarbonate water, and the pH value of the blue returning liquid is 1.5-11.0;
the eosin dye solution comprises 1-10 parts of eosin Y, 0-950 parts of ethanol, 0-4 parts of glacial acetic acid and 50-1000 parts of water, and the pH value of the eosin dye solution is 2.5-5.0.
2. The method for preparing a staining kit for pathological tissue sections according to claim 1, wherein the staining kit comprises: in the stabilizing solution, the organic acid comprises glacial acetic acid, tartaric acid and citric acid, and the polyalcohol comprises ethanol, ethylene glycol and propylene glycol.
3. The method for preparing a staining kit for pathological tissue sections according to claim 1, wherein the staining kit comprises: in the hematoxylin staining solution, the polyhydric alcohol comprises ethanol, ethylene glycol, propylene glycol and glycerol, the oxidizing agent comprises sodium iodate and potassium iodate, the mordant comprises aluminum sulfate, potassium aluminum sulfate and ammonium aluminum sulfate, and the organic acid comprises glacial acetic acid and citric acid.
4. The method for preparing a staining kit for pathological tissue section according to claim 1, wherein the staining kit comprises: in the differentiation solution, the acid includes hydrochloric acid, tartaric acid and glacial acetic acid, and the polyhydric alcohol includes ethanol, ethylene glycol and propylene glycol.
5. The method for preparing a staining kit for pathological tissue sections according to claim 1, wherein the staining kit comprises: in the bluing solution, the alkaline compounds comprise ammonia water, sodium bicarbonate, lithium carbonate, magnesium sulfate and tris (hydroxymethyl) aminomethane, and the polyols comprise ethylene glycol and propylene glycol.
6. The method for preparing a staining kit for pathological tissue sections according to claim 1, wherein the staining kit comprises: in the eosin dye solution, the eosin Y comprises alcohol-soluble eosin and water-soluble eosin.
7. A staining method of a staining kit for a pathological tissue section, comprising staining using the staining kit prepared by the method for preparing a staining kit for a pathological tissue section according to any one of claims 1 to 6.
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