Environment-friendly hematoxylin staining solution formula

Abstract

The invention discloses an environment-friendly hematoxylin staining solution formula, which is prepared by adding pure water into the following components according to the formula ratio: 5g / L of hematoxylin; 250 mL of 95% ethanol; 200 mL / L of glycerol; 200 mL / L of 1, 2-propylene glycol; 30 g / L of aluminum sulfate; and 0.8 g / L of sodium iodate. According to the formula of the environment-friendly hematoxylin staining solution, various raw materials are non-toxic and harmless substances, so that the harm of chemicals and toxic heavy metals to a human body is greatly reduced, the purpose of reducing the harm of an oxidized hematoxylin staining reagent to the human body is achieved, the environment-friendly and safe effects are achieved, meanwhile, the types of reagents are reduced, and the cost is reduced. The hematoxylin staining reagent is simpler in preparation, more convenient in staining operation, free of influence on staining effect, high in staining capacity and long in storage period, can be used for staining secretions, body fluids, body cavities and body surface exfoliated cell smears, and has the purpose of reducing harm of the hematoxylin staining reagent to human bodies. The method can be widely used in the staining fields of histology, cytology and the like.

Description

Technical Field

[0001] The invention relates to the technical fields of histology, cytology and the like, and in particular to a formula of an environmentally friendly hematoxylin staining solution. Background Art

[0002] Hematoxylin is an alkaline dye that can be used with a mordant (commonly trivalent iron or aluminum salts) to stain cell nuclei. Neither hematoxylin nor oxidized hematoxylin has staining ability, but forms a color precipitate (lake) when combined with a mordant. This lake has staining ability. Nuclei, chromosomes, centromeres, centrosomes, mitochondria, myelin sheaths, etc. can be stained blue or even black. The hematoxylin-eosin double staining method used in combination with eosin is the most basic histological and cytological staining method.

[0003] The nucleus in the cell is composed of acidic substances, which have a strong affinity with alkaline dyes (hematoxylin). Therefore, after the cell or tissue section is stained with hematoxylin staining solution, the nucleus is stained with hematoxylin into a bright blue-purple color. Almost all conventional sections, frozen sections, smears, and counterstaining after immunohistochemistry use this staining solution, which has many advantages, but also has unsatisfactory aspects, such as the complex preparation process, the many precautions for the preparation method, and the difficulty in controlling it. Moreover, mercuric oxide is a recognized highly toxic and harmful chemical with certain toxicity, which is easy to cause environmental pollution and adverse effects on human health. Therefore, we proposed a formula for an environmentally friendly hematoxylin staining solution. Summary of the invention

[0004] The main purpose of the present invention is to provide an environmentally friendly hematoxylin staining solution formula, 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] An environmentally friendly hematoxylin staining solution formula is prepared by adding the following components in the following formula amounts to pure water:

[0007] Hematoxylin 2-5 g / L;

[0008] 95% ethanol 150-250mL;

[0009] Glycerol 100-200mL / L;

[0010] 1,2-propylene glycol 150-200 mL / L;

[0011] Aluminum sulfate 15-30g / L;

[0012] Sodium iodate 0.5-0.8g / L.

[0013] Preferably, it is prepared by adding purified water to the following ingredients:

[0014] Hematoxylin 3 g / L;

[0015] 95% ethanol 200mL;

[0016] Glycerol 150mL / L;

[0017] 1,2-propylene glycol 170 mL / L;

[0018] Aluminum sulfate 22g / L;

[0019] Sodium iodate 0.6g / L.

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

[0021] S1: adding hematoxylin to a mixed solution of 95% ethanol, glycerol and 1,2-propylene glycol, and stirring to obtain a hematoxylin alcohol solution;

[0022] S2: adding aluminum sulfate to pure water and stirring to dissolve to obtain an aluminum sulfate solution;

[0023] S3: Add the hematoxylin alcohol solution into the aluminum sulfate solution, and then add sodium iodate, stir and mix well, and then add pure water to obtain an environmentally friendly hematoxylin staining solution.

[0024] Preferably, the environmentally friendly hematoxylin staining solution should be stored away from light in a 5°C-25°C environment with a relative humidity of no more than 80%, no corrosive gases and good ventilation.

[0025] Preferably, in S1, the stirring speed is 220-260 rpm, and the stirring time is 6-10 min.

[0026] Preferably, in S2, the stirring speed is 500-800 rpm, and the stirring time is 2-3 min.

[0027] Preferably, in S3, the stirring speed is 300-350 rpm, and the stirring time is 5-8 min.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] 1. The present invention artificially oxidizes hematoxylin by adding raw materials such as sodium iodate, so that hematoxylin undergoes oxidation to remove two hydrogen atoms and is converted into hematoxylin red, thereby providing a material basis for the effect of hematoxylin in staining cell nuclei. The staining function of hematoxylin can be achieved without the use of yellow mercuric oxide oxidation, and various raw materials are non-toxic and harmless substances, which greatly reduces the harm caused by chemicals and toxic heavy metals to the human body, achieves the purpose of reducing the harm caused by oxidized hematoxylin staining reagents to the human body, and has environmental protection and safety effects.

[0030] 2. The present invention reduces the types of reagents, makes reagent preparation simpler, and makes dyeing operation more convenient. At the same time, the dyeing effect is not affected. The synergistic effect between isopropanol and ethanol is utilized to increase the dehydration and transparency effects of ethanol, thereby achieving better dyeing effect.

[0031] 3. The preparation method of the hematoxylin staining solution of the present invention is simple, the preparation process does not require heating, the dyeing process is safe, the dyeing operation is simple, the dyeing ability is strong, and the storage period is long. It can be used to dye secretions, body fluids, body cavities, and body surface exfoliated cell smears, and has the purpose of reducing the harm of the hematoxylin staining reagent to the human body. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 The present invention is a flow chart of the preparation of an environmentally friendly hematoxylin staining solution formula;

[0033] Figure 2 This is a microscopic photograph of a smear of exfoliated cells on the body surface stained with an environmentally friendly hematoxylin staining solution (magnification 200 times). DETAILED DESCRIPTION

[0034] 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.

[0035] An environmentally friendly hematoxylin staining solution formula is prepared by adding the following components in the following formula amounts to pure water:

[0036] Hematoxylin 2-5 g / L;

[0037] 95% ethanol 150-250mL;

[0038] Glycerol 100-200mL / L;

[0039] 1,2-propylene glycol 150-200 mL / L;

[0040] Aluminum sulfate 15-30g / L;

[0041] Sodium iodate 0.5-0.8g / L.

[0042] like Figure 1 As shown, the following preparation steps are also included:

[0043] S1: Add hematoxylin to a mixed solution of 95% ethanol, glycerol and 1,2-propylene glycol, and stir to obtain a hematoxylin alcohol solution. The stirring speed is 220-260 rpm and the stirring time is 6-10 min. High concentration of ethanol can dehydrate and fix cells, making the cell environment in the tissue similar to that of the tissue, which is conducive to the coloring of cells in the tissue. There is a synergistic effect between isopropanol and ethanol, which can increase the dehydration and transparency of ethanol, thereby achieving better staining effect.

[0044] S2: adding aluminum sulfate to pure water and stirring to dissolve to obtain an aluminum sulfate solution, the stirring speed is 500-800 rpm, and the stirring time is 2-3 min;

[0045] S3: Add the alcohol solution of hematoxylin to the aluminum sulfate solution, and then add sodium iodate, stir and mix, then add pure water to obtain an environmentally friendly hematoxylin staining solution. The stirring speed is 300-350 rpm, and the stirring time is 5-8 min. Sodium iodate and other raw materials are added to artificially oxidize the hematoxylin, so that the hematoxylin undergoes oxidation to remove two hydrogen atoms and is converted into hematoxylin red, thereby providing a material basis for the hematoxylin to stain cell nuclei.

[0046] The environmentally friendly hematoxylin staining solution should be stored away from light in a 5°C-25°C environment with a relative humidity of no more than 80%, no corrosive gases and good ventilation.

[0047] Example 1

[0048] An environmentally friendly hematoxylin staining solution formula is prepared by adding the following components in the following formula amounts to pure water:

[0049] Hematoxylin 3 g / L;

[0050] 95% ethanol 200mL;

[0051] Glycerol 150mL / L;

[0052] 1,2-propylene glycol 170 mL / L;

[0053] Aluminum sulfate 22g / L;

[0054] Sodium iodate 0.6g / L.

[0055] The following preparation steps are also included:

[0056] S1: adding hematoxylin to a mixed solution of 95% ethanol, glycerol and 1,2-propylene glycol, stirring and mixing to obtain a hematoxylin alcohol solution, the stirring speed is 220-260 rpm, and the stirring time is 6-10 min;

[0057] S2: adding aluminum sulfate to pure water and stirring to dissolve to obtain an aluminum sulfate solution, the stirring speed is 500-800 rpm, and the stirring time is 2-3 min;

[0058] S3: Add the hematoxylin alcohol solution to the aluminum sulfate solution, and then add sodium iodate, stir and mix well, then add pure water to obtain an environmentally friendly hematoxylin staining solution, with a stirring speed of 300-350 rpm and a stirring time of 5-8 min.

[0059] The environmentally friendly hematoxylin staining solution should be stored away from light in a 5°C-25°C environment with a relative humidity of no more than 80%, no corrosive gases and good ventilation.

[0060] The staining solution of this embodiment only takes 3 minutes to use, and the staining solution can be used continuously for six months. Figure 2 The cell nucleus was clearly stained, and the chromatin, nuclear membrane and nucleolus structures were clear.

[0061] Example 2

[0062] An environmentally friendly hematoxylin staining solution formula is prepared by adding the following components in the following formula amounts to pure water:

[0063] Hematoxylin 2 g / L;

[0064] 95% ethanol 150mL;

[0065] Glycerol 100mL / L;

[0066] 1,2-propylene glycol 150 mL / L;

[0067] Aluminum sulfate 15g / L;

[0068] Sodium iodate 0.5g / L.

[0069] The following preparation steps are also included:

[0070] S1: adding hematoxylin to a mixed solution of 95% ethanol, glycerol and 1,2-propylene glycol, stirring and mixing to obtain a hematoxylin alcohol solution, the stirring speed is 220-260 rpm, and the stirring time is 6-10 min;

[0071] S2: adding aluminum sulfate to pure water and stirring to dissolve to obtain an aluminum sulfate solution, the stirring speed is 500-800 rpm, and the stirring time is 2-3 min;

[0072] S3: Add the hematoxylin alcohol solution to the aluminum sulfate solution, and then add sodium iodate, stir and mix well, then add pure water to obtain an environmentally friendly hematoxylin staining solution, with a stirring speed of 300-350 rpm and a stirring time of 5-8 min.

[0073] The environmentally friendly hematoxylin staining solution should be stored away from light in a 5°C-25°C environment with a relative humidity of no more than 80%, no corrosive gases and good ventilation.

[0074] The time of using the staining solution of this embodiment is only 6 minutes, and the staining solution can be used continuously for six months. The tissue section staining experiment proves that the cell nucleus is clearly stained, and the chromatin, nuclear membrane and nucleolus structures are clear.

[0075] Example 3

[0076] An environmentally friendly hematoxylin staining solution formula is prepared by adding the following components in the following formula amounts to pure water:

[0077] Hematoxylin 5 g / L;

[0078] 95% ethanol 250mL;

[0079] Glycerol 200mL / L;

[0080] 1,2-propylene glycol 200 mL / L;

[0081] Aluminum sulfate 30g / L;

[0082] Sodium iodate 0.8g / L.

[0083] The following preparation steps are also included:

[0084] S1: adding hematoxylin to a mixed solution of 95% ethanol, glycerol and 1,2-propylene glycol, stirring and mixing to obtain a hematoxylin alcohol solution, the stirring speed is 220-260 rpm, and the stirring time is 6-10 min;

[0085] S2: adding aluminum sulfate to pure water and stirring to dissolve to obtain an aluminum sulfate solution, the stirring speed is 500-800 rpm, and the stirring time is 2-3 min;

[0086] S3: Add the hematoxylin alcohol solution to the aluminum sulfate solution, and then add sodium iodate, stir and mix well, then add pure water to obtain an environmentally friendly hematoxylin staining solution, with a stirring speed of 300-350 rpm and a stirring time of 5-8 min.

[0087] The environmentally friendly hematoxylin staining solution should be stored away from light in a 5°C-25°C environment with a relative humidity of no more than 80%, no corrosive gases and good ventilation.

[0088] The time of using the staining solution of this embodiment is only 4 minutes, and the staining solution can be used continuously for six months. The tissue section staining experiment proves that the cell nucleus is clearly stained, and the chromatin, nuclear membrane and nucleolus structures are clear.

[0089] The environmentally friendly hematoxylin staining liquid formula of the present invention artificially oxidizes hematoxylin by adding raw materials such as sodium iodate, so that hematoxylin undergoes oxidation to remove two hydrogen atoms and is converted into hematoxylin red, thereby providing a material basis for the hematoxylin to stain cell nuclei, and the staining function of hematoxylin can be achieved without using yellow mercuric oxide oxidation, and various raw materials are non-toxic and harmless substances, which greatly reduces the harm caused by chemicals and toxic heavy metals to the human body, achieves the purpose of reducing the harm caused by oxidized hematoxylin staining reagents to the human body, and has environmental protection and safety effects; the types of reagents are reduced, the reagent preparation is simpler, the staining operation is more convenient, and the staining effect is not affected at the same time, and the synergistic effect between isopropanol and ethanol is utilized to increase the dehydration and transparency effects of ethanol, so as to achieve better staining effect; the preparation method of the hematoxylin staining liquid is simple, the preparation process does not need to be heated, the staining process is safe, the staining operation is simple, the dyeing liquid has strong staining ability, and the storage period is long, and the hematoxylin staining liquid can be used to stain secretions, body fluids, body cavities, and body surface exfoliated cell smears, and has the purpose of reducing the harm caused by the hematoxylin staining reagent to the human body.

[0090] 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. An environmentally friendly hematoxylin staining solution formula, characterized in that: It is prepared by adding pure water to the following ingredients: Hematoxylin 2-5 g / L; 95% ethanol 150-250mL; Glycerol 100-200mL / L; 1,2-propylene glycol 150-200 mL / L; Aluminum sulfate 15-30g / L; Sodium iodate 0.5-0.8g / L.

2. The environmentally friendly hematoxylin staining solution according to claim 1, characterized in that: It is prepared by adding the following ingredients and pure water: Hematoxylin 3 g / L; 95% ethanol 200mL; Glycerol 150mL / L; 1,2-propylene glycol 170 mL / L; Aluminum sulfate 22g / L; Sodium iodate 0.6g / L.

3. The environmentally friendly hematoxylin staining solution according to claim 1, characterized in that: The following preparation steps are also included: S1: adding hematoxylin to a mixed solution of 95% ethanol, glycerol and 1,2-propylene glycol, and stirring to obtain a hematoxylin alcohol solution; S2: adding aluminum sulfate to pure water and stirring to dissolve to obtain an aluminum sulfate solution; S3: Add the hematoxylin alcohol solution into the aluminum sulfate solution, and then add sodium iodate, stir and mix well, and then add pure water to obtain an environmentally friendly hematoxylin staining solution.

4. The environmentally friendly hematoxylin staining solution according to claim 1, characterized in that: The environmentally friendly hematoxylin staining solution should be stored away from light in a 5°C-25°C environment with a relative humidity of no more than 80%, no corrosive gases and good ventilation.

5. The environmentally friendly hematoxylin staining solution according to claim 3, characterized in that: In S1, the stirring speed is 220-260 rpm, and the stirring time is 6-10 min.

6. The environmentally friendly hematoxylin staining solution according to claim 3, characterized in that: In the step S2, the stirring speed is 500-800 rpm, and the stirring time is 2-3 min.

7. The environmentally friendly hematoxylin staining solution according to claim 3, characterized in that: In S3, the stirring speed is 300-350 rpm, and the stirring time is 5-8 min.

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