CN113218731A - Hematoxylin staining solution and preparation method thereof - Google Patents
Hematoxylin staining solution and preparation method thereof Download PDFInfo
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- CN113218731A CN113218731A CN202110548051.8A CN202110548051A CN113218731A CN 113218731 A CN113218731 A CN 113218731A CN 202110548051 A CN202110548051 A CN 202110548051A CN 113218731 A CN113218731 A CN 113218731A
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- hematoxylin
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- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 title claims abstract description 204
- 239000012192 staining solution Substances 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title abstract description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 78
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 68
- DIZPMCHEQGEION-UHFFFAOYSA-H aluminium sulfate (anhydrous) Chemical compound [Al+3].[Al+3].[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O.[O-]S([O-])(=O)=O DIZPMCHEQGEION-UHFFFAOYSA-H 0.000 claims abstract description 45
- 239000000243 solution Substances 0.000 claims abstract description 28
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000012153 distilled water Substances 0.000 claims abstract description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000007800 oxidant agent Substances 0.000 claims abstract description 17
- 239000011259 mixed solution Substances 0.000 claims abstract description 16
- 230000001590 oxidative effect Effects 0.000 claims abstract description 14
- 238000010438 heat treatment Methods 0.000 claims abstract description 12
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 239000000706 filtrate Substances 0.000 claims abstract description 6
- 238000005259 measurement Methods 0.000 claims abstract description 6
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 238000003756 stirring Methods 0.000 claims abstract description 6
- 238000005303 weighing Methods 0.000 claims abstract description 6
- 230000001105 regulatory effect Effects 0.000 claims abstract description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 33
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 14
- JLKDVMWYMMLWTI-UHFFFAOYSA-M potassium iodate Chemical compound [K+].[O-]I(=O)=O JLKDVMWYMMLWTI-UHFFFAOYSA-M 0.000 claims description 12
- 235000006666 potassium iodate Nutrition 0.000 claims description 12
- 239000001230 potassium iodate Substances 0.000 claims description 12
- 229940093930 potassium iodate Drugs 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 8
- 229960000583 acetic acid Drugs 0.000 claims description 7
- 239000012362 glacial acetic acid Substances 0.000 claims description 7
- NALMPLUMOWIVJC-UHFFFAOYSA-N n,n,4-trimethylbenzeneamine oxide Chemical compound CC1=CC=C([N+](C)(C)[O-])C=C1 NALMPLUMOWIVJC-UHFFFAOYSA-N 0.000 claims description 6
- 235000015281 sodium iodate Nutrition 0.000 claims description 6
- 239000011697 sodium iodate Substances 0.000 claims description 6
- 229940032753 sodium iodate Drugs 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 4
- 229960005150 glycerol Drugs 0.000 description 18
- 238000010186 staining Methods 0.000 description 14
- 238000004043 dyeing Methods 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 229910052751 metal Inorganic materials 0.000 description 7
- 239000002184 metal Substances 0.000 description 7
- 210000003855 cell nucleus Anatomy 0.000 description 6
- 229940093476 ethylene glycol Drugs 0.000 description 5
- 229960004106 citric acid Drugs 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 108010077544 Chromatin Proteins 0.000 description 3
- 210000003483 chromatin Anatomy 0.000 description 3
- 230000003647 oxidation Effects 0.000 description 3
- 238000007254 oxidation reaction Methods 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 239000003381 stabilizer Substances 0.000 description 3
- 230000032683 aging Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 210000000805 cytoplasm Anatomy 0.000 description 2
- 239000000975 dye Substances 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- -1 pH regulator Chemical compound 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000007447 staining method Methods 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 229930182559 Natural dye Natural products 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 239000000980 acid dye Substances 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000005562 fading Methods 0.000 description 1
- 230000000887 hydrating effect Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000978 natural dye Substances 0.000 description 1
- 238000012758 nuclear staining Methods 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B61/00—Dyes of natural origin prepared from natural sources, e.g. vegetable sources
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B65/00—Compositions containing mordants
-
- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09B—ORGANIC DYES OR CLOSELY-RELATED COMPOUNDS FOR PRODUCING DYES, e.g. PIGMENTS; MORDANTS; LAKES
- C09B67/00—Influencing the physical, e.g. the dyeing or printing properties of dyestuffs without chemical reactions, e.g. by treating with solvents grinding or grinding assistants, coating of pigments or dyes; Process features in the making of dyestuff preparations; Dyestuff preparations of a special physical nature, e.g. tablets, films
- C09B67/0071—Process features in the making of dyestuff preparations; Dehydrating agents; Dispersing agents; Dustfree compositions
- C09B67/0072—Preparations with anionic dyes or reactive dyes
- C09B67/0073—Preparations of acid or reactive dyes in liquid form
-
- 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 discloses a hematoxylin dye solution and a preparation method thereof, wherein the hematoxylin dye solution comprises hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, an oxidant, glycerol, a pH regulator and glycol; the preparation method comprises the following steps: s1, weighing hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, an oxidant, glycerol, a pH regulator and ethylene glycol according to the measurement; s2, sequentially adding absolute ethyl alcohol, ethylene glycol and hematoxylin into a beaker, and heating until the hematoxylin is completely dissolved to obtain a hematoxylin solution; s3, sequentially adding distilled water and aluminum sulfate into a container, and heating until the aluminum sulfate is completely dissolved to obtain an aluminum sulfate solution; s4, adding an aluminum sulfate solution into a hematoxylin solution, stirring, sequentially adding an oxidant and glycerol, fully mixing, adding a pH regulator, and regulating the pH value to 2-3.5; s5, standing for 12-24 hours, and filtering the mixed solution to obtain a filtrate, namely the hematoxylin staining solution.
Description
Technical Field
The invention relates to the field of biochemistry, and particularly relates to a hematoxylin dye solution and a preparation method thereof.
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 staining method mainly comprise slicing, baking, dewaxing, hydrating
Hematoxylin staining-differentiation-blueing-eosin staining-dehydration-transparency-sealing; the hematoxylin staining operation mainly uses a hematoxylin staining solution, and the following problems are common at present:
1) the hematoxylin staining solution is easy to generate a metal film on the surface, or the staining material precipitates, 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 incapability are caused.
2) For the metal membrane generated by hematoxylin staining solution, the metal membrane needs to be filtered and removed, which increases the workload of technicians and wastes time and reagents.
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 fading is easy, and the use requirement of 30-year storage cannot be met.
5) The 500ml hematoxylin staining solution can not stain 2000 pieces of slices generally, and the utilization rate is poor.
6) 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 workload is increased, and the efficiency is reduced.
Disclosure of Invention
The invention aims to solve the problems and provides hematoxylin staining solution for improving the nuclear staining effect and a preparation method thereof.
In order to achieve the purpose, the technical scheme of the invention is as follows:
a hematoxylin staining solution comprises hematoxylin, anhydrous ethanol, aluminum sulfate, distilled water, oxidant, glycerol, pH regulator, and ethylene glycol; the weight percentage of each component is as follows: 0.2% of hematoxylin, 15-20% of absolute ethyl alcohol, 1% of aluminum sulfate, 65-70% of distilled water, 0.01-0.02% of oxidant, 5% of glycerol, 0.2% of pH regulator and 7.7-7.9% of glycol.
Further, the hematoxylin staining solution comprises the following components in percentage by mass: 0.2 percent of hematoxylin, 17.79 percent of absolute ethyl alcohol, 1 percent of aluminum sulfate, 68 percent of distilled water, 0.01 percent of oxidant, 5 percent of glycerol, 0.2 percent of PH regulator and 7.8 percent of glycol.
Further, the oxidizing agent is one or a mixture of potassium iodate and sodium iodate.
Further, the pH regulator is one or a mixture of citric acid and glacial acetic acid.
A preparation method of hematoxylin staining solution comprises the following steps:
s1, weighing hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, an oxidant, glycerol, a pH regulator and ethylene glycol according to the measurement;
s2, sequentially adding absolute ethyl alcohol, ethylene glycol and hematoxylin into a beaker, and heating until the hematoxylin is completely dissolved to obtain a hematoxylin solution;
s3, sequentially adding distilled water and aluminum sulfate into a container, and heating until the aluminum sulfate is completely dissolved to obtain an aluminum sulfate solution;
s4, adding an aluminum sulfate solution into a hematoxylin solution, stirring uniformly, sequentially adding an oxidant and glycerol, fully and uniformly mixing to obtain a mixed solution, adding a pH regulator into the mixed solution, and regulating the pH value to 2-3.5;
s5, standing for 12-24 hours, and filtering the mixed solution to obtain a filtrate, namely the hematoxylin staining solution.
Further, a filter screen of 1000 to 2000 meshes is used for filtering in the step S5.
Compared with the prior art, the invention has the advantages and positive effects that:
1) the hematoxylin staining solution disclosed by the invention has the advantages that by adjusting the proportion of metal salts in the components, no metal film is generated in the using process, the workload of technical personnel is greatly reduced, and the staining efficiency is improved.
2) The hematoxylin staining solution disclosed by the invention is matched with a stabilizer for use, so that the hematoxylin staining solution has stronger selectivity to cell nuclei, clearer and more distinct staining effect and longer preservation time, and can meet the long-term preservation requirement through an aging test (60 ℃ and 24h 7).
3) The hematoxylin staining solution can stain at least 3000 slices per 500ml, and the utilization rate of the hematoxylin staining solution is effectively improved.
4) The sensitivity of the hematoxylin staining solution to the temperature is greatly reduced, the staining time does not need to be groped and adjusted due to the temperature change, and the working efficiency is improved.
5) The hematoxylin staining solution disclosed by the invention is free from oxide film formation, can be used for a long time, and meanwhile, nuclear chromatin is deeply and finely stained, so that the staining effect is enhanced.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to 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 from the embodiments of the present invention by a person skilled in the art without any creative effort, should be included in the protection scope of the present invention.
The components of the hematoxylin staining solution comprise hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, potassium iodate, glycerol, citric acid and glycol; the weight percentage of each component is as follows: 0.2 percent of hematoxylin, 20 percent of absolute ethyl alcohol, 1 percent of aluminum sulfate, 65.69 percent of distilled water, 0.01 percent of potassium iodate, 5 percent of glycerol, 0.2 percent of citric acid and 7.9 percent of glycol.
The preparation method of the hematoxylin staining solution comprises the following steps:
s1, weighing hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, potassium iodate, glycerol, citric acid and ethylene glycol according to the measurement;
s2, sequentially adding absolute ethyl alcohol, ethylene glycol and hematoxylin into a beaker, and heating until the hematoxylin is completely dissolved to obtain a hematoxylin solution;
s3, sequentially adding distilled water and aluminum sulfate into a container, and heating until the aluminum sulfate is completely dissolved to obtain an aluminum sulfate solution;
s4, adding an aluminum sulfate solution into a hematoxylin solution, stirring uniformly, sequentially adding potassium iodate and glycerol, fully mixing to obtain a mixed solution, adding citric acid into the mixed solution, and adjusting the pH value to 2;
s5, standing for 12 hours, and filtering the mixed solution by using a 1000-mesh filter screen to obtain a filtrate, namely the hematoxylin staining solution.
Example 2, this example discloses a hematoxylin staining solution, the components of which include hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, sodium iodate, glycerol, glacial acetic acid, and ethylene glycol; the weight percentage of each component is as follows: 0.2 percent of hematoxylin, 17.79 percent of absolute ethyl alcohol, 1 percent of aluminum sulfate, 68 percent of distilled water, 0.01 percent of sodium iodate, 5 percent of glycerol, 0.2 percent of glacial acetic acid and 7.8 percent of glycol.
The preparation method of the hematoxylin staining solution comprises the following steps:
s1, weighing hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, sodium iodate, glycerol, glacial acetic acid and ethylene glycol according to the measurement;
s2, sequentially adding absolute ethyl alcohol, ethylene glycol and hematoxylin into a beaker, and heating until the hematoxylin is completely dissolved to obtain a hematoxylin solution;
s3, sequentially adding distilled water and aluminum sulfate into a container, and heating until the aluminum sulfate is completely dissolved to obtain an aluminum sulfate solution;
s4, adding an aluminum sulfate solution into a hematoxylin solution, stirring uniformly, sequentially adding potassium iodate and glycerol, fully mixing to obtain a mixed solution, adding glacial acetic acid into the mixed solution, and adjusting the pH value to 2;
s5, standing for 18 hours, and filtering the mixed solution by using a 2000-mesh filter screen to obtain a filtrate, namely the hematoxylin staining solution.
Example 3, this example discloses a hematoxylin staining solution, wherein the components of the hematoxylin staining solution include hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, potassium iodate, glycerol, citric acid, and ethylene glycol; the weight percentage of each component is as follows: 0.2 percent of hematoxylin, 15.68 percent of absolute ethyl alcohol, 1 percent of aluminum sulfate, 70 percent of distilled water, 0.02 percent of potassium iodate, 5 percent of glycerol, 0.2 percent of citric acid and 7.9 percent of glycol.
The preparation method of the hematoxylin staining solution comprises the following steps:
s1, weighing hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, potassium iodate, glycerol, citric acid and ethylene glycol according to the measurement;
s2, sequentially adding absolute ethyl alcohol, ethylene glycol and hematoxylin into a beaker, and heating until the hematoxylin is completely dissolved to obtain a hematoxylin solution;
s3, sequentially adding distilled water and aluminum sulfate into a container, and heating until the aluminum sulfate is completely dissolved to obtain an aluminum sulfate solution;
s4, adding an aluminum sulfate solution into a hematoxylin solution, stirring uniformly, sequentially adding potassium iodate and glycerol, fully mixing to obtain a mixed solution, adding citric acid into the mixed solution, and adjusting the pH value to 3.5;
s5, standing for 24 hours, and filtering the mixed solution by using a 2000-mesh filter screen to obtain a filtrate, namely the hematoxylin staining solution.
The component analysis in the invention:
the hematoxylin is a natural dye, is not a dye (only has a chromophoric hydroxyl group) per se, has no dyeing performance, and has dyeing capability only after being oxidized into oxidized hematoxylin (hematoxylin) which has the chromophoric hydroxyl group and a quinoid chromophore.
The anhydrous ethanol and ethylene glycol are mainly used as solvents for dissolving hematoxylin and inhibiting the growth of mold.
Aluminum sulfate is mainly used as a mordant, aluminum ions are chelated with hematoxylin to form blue precipitates (with positive charges), and the blue precipitates are in polar combination with phosphate radicals (with negative charges) of nucleic acid of cell nuclei, so that the cell nuclei are finally colored.
Potassium iodate or sodium iodate is mainly used as an oxidizing agent, and can artificially accelerate the oxidation of hematoxylin.
The glycerol is mainly used as a stabilizer, so that the condition of excessive oxidation of the hematoxylin is avoided.
Citric acid or glacial acetic acid is used for balancing the oxidation of the oxidizing agent and increasing the selective staining effect of cell nucleus.
Distilled water is mainly used as a solvent for dissolving aluminum sulfate.
The invention has the following beneficial effects:
1. the hematoxylin staining solution disclosed by the invention has the advantages that by adjusting the proportion of metal salts in the components, no metal film is generated in the using process, the workload of technical personnel is greatly reduced, and the staining efficiency is improved.
2. The hematoxylin staining solution disclosed by the invention is matched with a stabilizer for use, so that the hematoxylin staining solution has stronger selectivity to cell nuclei, clearer and more distinct staining effect and longer preservation time, and can meet the long-term preservation requirement through an aging test (60 ℃ and 24h 7).
3. The hematoxylin staining solution can stain at least 3000 slices per 500ml, and the utilization rate of the hematoxylin staining solution is effectively improved.
4. The sensitivity of the hematoxylin staining solution to the temperature is greatly reduced, the staining time does not need to be groped and adjusted due to the temperature change, and the working efficiency is improved.
5. The hematoxylin staining solution disclosed by the invention is free from oxide film formation, can be used for a long time, and meanwhile, nuclear chromatin is deeply and finely stained, so that the staining effect is enhanced.
Claims (6)
1. A hematoxylin staining solution is characterized in that: the hematoxylin staining solution comprises hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, an oxidant, glycerol, a pH regulator and glycol; the weight percentage of each component is as follows: 0.2% of hematoxylin, 15-20% of absolute ethyl alcohol, 1% of aluminum sulfate, 65-70% of distilled water, 0.01-0.02% of oxidant, 5% of glycerol, 0.2% of pH regulator and 7.7-7.9% of glycol.
2. The hematoxylin stain of claim 1, wherein: the hematoxylin staining solution comprises the following components in percentage by mass: 0.2 percent of hematoxylin, 17.79 percent of absolute ethyl alcohol, 1 percent of aluminum sulfate, 68 percent of distilled water, 0.01 percent of oxidant, 5 percent of glycerol, 0.2 percent of PH regulator and 7.8 percent of glycol.
3. The hematoxylin stain of either claim 1 or claim 2, wherein: the oxidant is one or the mixture of two of potassium iodate and sodium iodate.
4. The hematoxylin stain of claim 3, wherein: the pH regulator is one or a mixture of citric acid and glacial acetic acid.
5. The method for preparing the hematoxylin stain of claim 4, wherein the method comprises the following steps: the method comprises the following steps:
s1, weighing hematoxylin, absolute ethyl alcohol, aluminum sulfate, distilled water, an oxidant, glycerol, a pH regulator and ethylene glycol according to the measurement;
s2, sequentially adding absolute ethyl alcohol, ethylene glycol and hematoxylin into a beaker, and heating until the hematoxylin is completely dissolved to obtain a hematoxylin solution;
s3, sequentially adding distilled water and aluminum sulfate into a container, and heating until the aluminum sulfate is completely dissolved to obtain an aluminum sulfate solution;
s4, adding an aluminum sulfate solution into a hematoxylin solution, stirring uniformly, sequentially adding an oxidant and glycerol, fully and uniformly mixing to obtain a mixed solution, adding a pH regulator into the mixed solution, and regulating the pH value to 2-3.5;
s5, standing for 12-24 hours, and filtering the mixed solution to obtain a filtrate, namely the hematoxylin staining solution.
6. The method for preparing hematoxylin stain according to claim 5, wherein: and in the step S5, a filter screen with 1000-2000 meshes is used for filtering.
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| CN202110548051.8A CN113218731A (en) | 2021-05-19 | 2021-05-19 | Hematoxylin staining solution and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113933133A (en) * | 2021-10-25 | 2022-01-14 | 易普森生物科技(深圳)有限公司 | Staining solution for cell sample |
| CN114624084A (en) * | 2022-03-18 | 2022-06-14 | 深圳市贝安特医疗技术有限公司 | Preparation and staining method of pathological tissue section staining kit |
-
2021
- 2021-05-19 CN CN202110548051.8A patent/CN113218731A/en not_active Withdrawn
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113933133A (en) * | 2021-10-25 | 2022-01-14 | 易普森生物科技(深圳)有限公司 | Staining solution for cell sample |
| CN114624084A (en) * | 2022-03-18 | 2022-06-14 | 深圳市贝安特医疗技术有限公司 | Preparation and staining method of pathological tissue section staining kit |
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