Lelfusin staining method kit and application thereof

Abstract

The present invention relates to the technical field of biology, and discloses a lefuchsin staining method kit, which comprises a lefuchsin staining solution and a collagenous fiber staining method staining solution, the solution is prepared from the following raw materials by mass: 2-20 g of basic fuchsin, 4-40 g of resorcin, 29-35 g of ferric trichloride, 5-20 g of hematoxylin, 1-20 g of acid fuchsin, 2.5-10 g of picric acid, 3-10 ml of hydrochloric acid, and 2000 ml of pure water. When the staining solution of the kit is used for staining, no staining solution precipitation residue exists, no oxidation film exists on the staining solution, and the elastic fibers can be dyed into the elastic fibers with dark blue to black blue, the collagenous fibers are red, the muscle fibers and red blood cells are yellow, and the cell nucleuses are black blue in an extremely short time.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and more specifically to a fuchsin staining kit and application thereof. Background Art

[0002] Elastic fibers are widely distributed in various parts of the body, among which the most abundant areas are the skin, blood vessel walls, ligaments, trachea, bronchi, alveoli, auricles, and glandular ducts. Elastic fibers are composed of elastin and are thin, shiny yellow lines when the specimen is fresh, so they are also called yellow fibers. Because they are elastic, they are usually called elastic fibers. They do not form a colloid when boiled in water, and have great resistance to acid and alkali. They are a hard protein that is extremely difficult to dissolve; they have strong refractive power and are easy to stretch; the fibers are spiral, with filamentous branches, and exist in the form of scattered connections to form a network or a thin layer. The length and thickness of the elastic fibers in each place are different, with a diameter of about 0.2 to 1 μm.

[0003] Special staining and histochemical staining are staining techniques developed on the basis of HE staining. They are one of the important auxiliary techniques in clinical pathological diagnosis. Many diseases that are difficult to diagnose by conventional HE staining can be further diagnosed by applying special staining and histochemical staining techniques. Although immunohistochemical technology is widely used at present, many special staining and histochemical staining cannot be replaced by immunohistochemical technology. The application of special staining and histochemical staining techniques helps to improve the level of clinical pathological diagnosis. Histochemical staining is a staining technique that combines histology, cytology and biochemistry. It uses certain reagents or dyes to react specifically with the chemical components in tissue cells or the molecular structure of the dye. Certain groups specifically bind to corresponding groups in tissues to develop color, which can not only determine certain special components of tissue cells, but also show the distribution and content of these special components in tissue cells and the changes that occur during lesions. With the development of pathology, simple HE staining can no longer meet the needs of clinical pathology for observation of tissue cell morphology. It is hoped that a new staining technology can solve the problems of observing the variation of tissue components when observing changes in tissue cell morphology, identifying certain similar substances or structures (such as pigments, cytoplasmic vacuoles, etc.) in tissues or cells seen in HE staining, and distinguishing pathogens in lesions. For this purpose, the present application proposes a fuchsin staining kit. Summary of the invention

[0004] The purpose of the present invention is to provide a fuchsin staining kit and application thereof in view of the deficiencies in the prior art.

[0005] The above technical purpose of the present invention is achieved through the following technical scheme: a fuchsin staining kit, including fuchsin staining solution and collagen fiber staining solution, the above solution is made of raw materials with the following mass components: basic fuchsin 2-20g, raclonidine 4-40g, ferric chloride 29-35g, hematoxylin 5-20g, acid fuchsin 1-20g, picric acid 2.5-10g, hydrochloric acid 3-10ml and pure water 2000ml.

[0006] Preferably, the method for preparing the fuchsin staining solution is to take 2-20g of basic fuchsin, 4-40g of ferric oxide and 200ml of pure water, add them into a beaker and heat to boil, then add 25ml of 29%-35% iron oxide aqueous solution, mix and stir with a glass stirring rod for 2-5min, cool to room temperature, then filter, pour off the filtrate, put the filter paper and the precipitate on it back into the beaker, and place them in a drying oven for drying, take out after drying and add 200ml of 95% alcohol solution to heat in a water bath until the precipitate is completely dissolved, take out the filter paper, filter after the liquid is cooled, add 95% alcohol solution to the solution to 200ml, and finally add 4ml-8ml of hydrochloric acid.

[0007] Preferably, the preparation method of the collagen fiber staining solution is to take 10-60 ml of 1%-20% by mass acid fuchsin aqueous solution, add 90-120 ml of 1.22% by mass saturated picric acid aqueous solution and mix well.

[0008] Preferably, the preparation process of the 29%-35% by mass ferric oxide aqueous solution is to add 29-35g of ferric chloride to 100ml of pure water and stir to mix evenly.

[0009] Preferably, the preparation method of the acid fuchsin aqueous solution with a mass fraction of 1%-20% is to add 1-20g of acid fuchsin into 100ml of pure water and stir to mix evenly.

[0010] According to the method for using the above-mentioned fuchsin staining kit, the method comprises the following steps: a. Paraffin sections 2-3 μm, dewaxed in 95% alcohol solution; b. Use Fuchsin staining solution for 1-2 hours; c. Remove the slices from the stain and wash off the excess stain with 95% alcohol solution, and differentiate until there is no residual color; d. Wash thoroughly with water for 5 minutes; e. Counterstain the cell nucleus with an aqueous solution of alum hematoxylin for 2-5 minutes; f. Wash with water for 2-3 minutes; g. Use collagen fiber staining method for contrast staining; h.95% alcohol solution rapidly differentiates; i. Dehydrate with anhydrous alcohol, make transparent with xylene, and seal with neutral gum.

[0011] For example, the above-mentioned fuchsin staining kit is used in elastic fibers, biological tissues and human breast cancer tissues.

[0012] Compared with the prior art, the solution of the present invention has the following beneficial effects: The dyeing solution of the kit of the present invention has no dyeing solution precipitation residue during dyeing, and no oxide film is formed on the dyeing solution. If an oxide film is generated, as the reagent continues to volatilize, the oxide film on the surface of the reagent adheres to the tissue section to form a dye precipitation, which is difficult to clean. At the same time, the elastic fibers can be dyed dark blue to black blue, the collagen fibers are red, the muscle fibers and red blood cells are yellow, and the nuclei are black blue in a very short time. DETAILED DESCRIPTION

[0013] The invention is further described in detail below through specific examples. The following examples are only illustrative and not limiting of the protection scope of the present invention.

[0014] Example 1 A fuchsin staining kit comprises fuchsin staining solution and collagen fiber staining solution, wherein the above solutions are prepared from raw materials of the following mass components: 2-20g of basic fuchsin, 4-40g of raclonidine, 29-35g of ferric chloride, 5-20g of hematoxylin, 1-20g of acid fuchsin, 2.5-10g of picric acid, 3-10ml of hydrochloric acid and 2000ml of pure water.

[0015] In this embodiment, the preparation method of the fuchsin staining solution is to take 2-20g of basic fuchsin, 4-40g of ferric oxide and 200ml of pure water, add them into a beaker and heat to boil, then add 25ml of 29%-35% iron oxide aqueous solution, mix and stir with a glass stirring rod for 2-5min, cool to room temperature, then filter, pour off the filtrate, put the filter paper and the precipitate thereon back into the beaker, and place them in a drying oven for drying, take out after drying and add 200ml of 95% alcohol solution to heat in a water bath until the precipitate is completely dissolved, take out the filter paper, filter after the liquid is cooled, add 95% alcohol solution to the solution to 200ml, and finally add 4ml-8ml of hydrochloric acid.

[0016] Furthermore, the preparation method of the collagen fiber staining method dyeing solution is to take 10-60 ml of an acid fuchsin aqueous solution with a mass fraction of 1%-20%, add 90-120 ml of a saturated aqueous solution of picric acid with a mass fraction of 1.22% and mix evenly. The preparation process of the 29%-35% ferric oxide aqueous solution is to take 29-35 g of ferric chloride and add it to 100 ml of pure water and stir and mix evenly. The preparation method of the 1%-20% acid fuchsin aqueous solution is to take 1-20 g of acid fuchsin and add it to 100 ml of pure water and stir and mix evenly.

[0017] The method for using the above-mentioned fuchsin staining kit comprises the following steps: a. Paraffin sections 2-3 μm, dewaxed in 95% alcohol solution; b. Use Fuchsin staining solution for 1-2 hours; c. Remove the slices from the stain and wash off the excess stain with 95% alcohol solution, and differentiate until there is no residual color; d. Wash thoroughly with water for 5 minutes; e. Counterstain the cell nucleus with an aqueous solution of alum hematoxylin for 2-5 minutes; f. Wash with water for 2-3 minutes; g. Use collagen fiber staining method for contrast staining; h.95% alcohol solution rapidly differentiates; i. Dehydrate with anhydrous alcohol, make transparent with xylene, and seal with neutral gum.

[0018] Example 2 In another embodiment of the present invention, the dye of the kit of the present invention is used for elastic fibers: in the fibrous tissue of elastic fiber tumors in biological tissues and in vitro specimens, there are many elastic fibers with spherical or granular changes, which are easily overlooked in HE staining. Elastic fiber staining can clearly show that there are abundant elastic fibers and elastic spheres in the tumor, thereby confirming the diagnosis.

[0019] Example 3 In another embodiment of the present invention, the dye of the kit of the present invention is used in biological tissues and human breast cancer tissues: it is seen around the ducts and blood vessel walls, and this phenomenon is often combined with scirrhous cancer and invasive lobular carcinoma; as the elastic fiber staining method proves in ductal carcinoma, accompanied by elastic fiber hyperplasia, it can be proved to be early invasive cancer.

[0020] This specific embodiment is only an explanation of the present invention, and it is not a limitation of the present invention. After reading this specification, those skilled in the art can make non-creative modifications to the implementation rules as needed, but they are protected by the patent law as long as they are within the scope of the claims of the present invention.

Claims

1. A fuchsin staining kit, characterized in that: The invention comprises a fuchsin staining solution and a collagen fiber staining method staining solution, wherein the above-mentioned solution is prepared from raw materials of the following mass components: 2-20g of basic fuchsin, 4-40g of raclonidine, 29-35g of ferric chloride, 5-20g of hematoxylin, 1-20g of acid fuchsin, 2.5-10g of picric acid, 3-10ml of hydrochloric acid and 2000ml of pure water.

2. A fuchsin staining kit according to claim 1, characterized in that, The preparation method of the fuchsin staining solution is as follows: 2-20g of basic fuchsin, 4-40g of ferric oxide and 200ml of pure water are added into a beaker for heating and boiling, and then 25ml of ferric oxide aqueous solution with a mass fraction of 29%-35% is added, mixed and stirred with a glass stirring rod for 2-5min, cooled to room temperature, and then filtered, the filtrate is poured off, the filter paper and the precipitate thereon are put back into the beaker together, and placed in a drying oven for drying, after drying, taken out and 200ml of 95% alcohol solution is added, and heated in a water bath until the precipitate is completely dissolved, the filter paper is taken out, the liquid is cooled and filtered, and then 95% alcohol solution is added to the solution to 200ml, and finally 4ml-8ml of hydrochloric acid is added.

3. A fuchsin staining kit according to claim 2, characterized in that, The preparation method of the collagen fiber dyeing solution is as follows: 10-60 ml of an acid fuchsin aqueous solution with a mass fraction of 1%-20% is taken, and 90-120 ml of a saturated picric acid aqueous solution with a mass fraction of 1.22% is added thereto and mixed evenly.

4. A fuchsin staining kit according to claim 3, characterized in that, The preparation process of the 29%-35% by mass ferric oxide aqueous solution is to add 29-35g of ferric chloride into 100ml of pure water and stir to mix evenly.

5. A fuchsin staining kit according to claim 4, characterized in that, The preparation method of the acid fuchsin aqueous solution with a mass fraction of 1%-20% is to take 1-20g of acid fuchsin and add it into 100ml of pure water and stir and mix evenly.

6. The method for using a fuchsin staining kit according to any one of claims 1 to 5, characterized in that: The following steps are included: a. Paraffin sections 2-3 μm, dewaxed in 95% alcohol solution; b. Use Fuchsin staining solution for 1-2 hours; c. Remove the slices from the stain and wash off the excess stain with 95% alcohol solution, and differentiate until there is no residual color; d. Wash thoroughly with water for 5 minutes; e. Counterstain the cell nucleus with an aqueous solution of alum hematoxylin for 2-5 minutes; f. Wash with water for 2-3 minutes; g. Use collagen fiber staining method for contrast staining; h.95% alcohol solution rapidly differentiates; i. Dehydrate with anhydrous alcohol, make transparent with xylene, and seal with neutral gum.

7. Use of a fuchsin staining kit as claimed in any one of claims 1 to 6 in elastic fibers, biological tissues and human breast cancer tissues.