Buffer decalcification stationary liquid

Abstract

The invention discloses a buffer decalcification stationary liquid, which is prepared by mixing a plurality of acidic materials, and the plurality of acidic materials comprise a lemon tree aqueous solution, ammonium citrate, zinc sulfate and trichloromethane. The bone tissue decalcifying agent is high in permeability and can remove moisture on the surfaces of cells, so that protein molecular structures are released, and proteins are denatured and coagulated to achieve a preservative effect. And meanwhile, the decalcification liquid does not have volatility, so that the relatively large defects on the safety of the environment and operators are overcome, and long-term storage of the decalcification liquid is facilitated.

Description

Technical Field

[0001] The invention relates to the technical field of living tissue pathological diagnosis, in particular to a buffered decalcification fixative. Background Art

[0002] Living tissue pathological diagnosis is the first diagnosis in surgery and the gold standard. Doctors can make a correct diagnosis based on the test results of the specimens and formulate the next treatment plan for the disease. The accuracy of the pathology report plays a decisive role in the diagnosis and treatment of the disease to a considerable extent. The correct handling of pathological specimens can not only provide effective guarantees for pathological diagnosis and clinical diagnosis, but also protect the patient's body and mind and eliminate disputes caused by improper specimen handling. Existing surveys show that the current specimen handling methods in the operating room cannot effectively prevent nursing errors. Due to changes in the microenvironment, autolysis and (or) corruption will occur, causing structural damage. At the same time, at the microscopic level, cells will degenerate and autolyze. The purpose of handling samples is to use various methods to keep the pathological specimens as close to their pre-ex vivo state as possible. More importantly, since microscopic observation is required, pathological histology not only requires the in vitro specimens to be as close to the pre-ex vivo state as possible in terms of macroscopic aspects (such as color, volume, hardness, etc.), but also requires them to be as close to the living state as possible at the microscopic level of the cell.

[0003] At present, the most common treatment agents for pathological specimens are decalcification agents such as nitric acid, hydrochloric acid, sulfuric acid, and EDTA decalcification solution:

[0004] 1. Nitric acid decalcification solution: It decalcifies quickly and has no tissue fixation effect. Nitric acid causes excessive damage to proteins and nucleic acids in tissues, seriously affecting the subsequent tissue staining effect and causing false negative results in subsequent immunohistochemistry and molecular pathology tests. Nitric acid is a strong acid with high requirements for transportation and storage. During transportation, storage and use, it poses a great threat to the safety of the environment and operators.

[0005] 2. Hydrochloric acid decalcification solution: decalcification is fast, and it does not have a tissue fixation effect. Hydrochloric acid causes too much damage to proteins and nucleic acids in tissues, affecting the subsequent tissue staining effect, and affecting the reliability of subsequent immunohistochemistry and molecular pathology test results. Hydrochloric acid is a strong acid, and has high requirements for transportation and storage. During transportation, storage and use, there are major safety hazards to the environment and the safety of operators.

[0006] 3. Sulfuric acid decalcification solution: It decalcifies quickly and has no tissue fixation effect. Sulfuric acid causes excessive damage to proteins and nucleic acids in tissues, seriously affecting the subsequent tissue staining effect and causing false negative results in subsequent immunohistochemistry and molecular pathology tests. Sulfuric acid is a strong acid with high requirements for transportation and storage. During transportation, storage and use, there are major safety hazards to the environment and the safety of operators.

[0007] 4. EDTA decalcification solution: The main component of EDTA decalcification solution is disodium EDTA, which causes little damage to tissues during decalcification and has little effect on subsequent staining and detection. However, because EDTA is a chelating agent that decalcifies through chelation reaction, the decalcification time takes 3 to 4 weeks, or even longer, and it is difficult to be promoted for use;

[0008] However, all of the above four methods have obvious defects. The above four decalcification solutions easily make the bone tissue specimens hard, deformed, faded, and not fresh. Summary of the invention

[0009] In view of the shortcomings of the prior art, the present invention provides a buffered decalcification fixative solution, which solves the problem mentioned in the background art that the current decalcification solution easily causes the bone tissue specimen to harden, deform, fade, and lose freshness.

[0010] To achieve the above objectives, the present invention is implemented through the following technical scheme: a buffered decalcification fixative, which is obtained by mixing a plurality of acidic substances together, wherein the plurality of acidic substances include lemon tree aqueous solution, ammonium citrate, zinc sulfate and chloroform.

[0011] In a further embodiment, the mass ratio of the lemon tree aqueous solution, ammonium citrate, zinc sulfate and chloroform is 5:95:0.2:3.

[0012] In a further embodiment, the concentration of the lemon tree aqueous solution per 5 ml is 7%.

[0013] In a further embodiment, the concentration of ammonium citrate per 95 ML is 7.54%.

[0014] In a further embodiment, the concentration of zinc sulfate per 0.2 ML is 1%.

[0015] A buffered decalcified fixative intended primarily for fixation of fresh bone tissue specimens.

[0016] The present invention provides a buffered decalcification fixative having the following beneficial effects:

[0017] It is a highly permeable bone tissue decalcification agent that can remove moisture from the cell surface, loosen the protein molecular structure, and denature and coagulate the protein to play an antiseptic role. At the same time, it is non-volatile, thus solving the major disadvantages of environmental and operator safety, and is conducive to the long-term storage of decalcified liquid. DETAILED DESCRIPTION

[0018] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with specific embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0019] The present invention provides a technical solution: a buffered decalcification fixative: mainly prepared by mixing 5ML of 7% lemon tree water solution, 95ML of 7.54% ammonium citrate, 0.2ML of 1% zinc sulfate, 3ML of chloroform, deionized water, etc., and the prepared buffered decalcification fixative can be used for fixing fresh bone tissue samples;

[0020] Compound decalcification fixative: mainly made of a mixture of formic acid, distilled water, acetic acid, formaldehyde, etc.

[0021] Buffered decalcification fluid can cause calcium salts inside bone tissue to react, turning them into calcium ions and softening the tissue, making it easier to perform conventional paraffin sections.

[0022] Clinical evaluation data of this buffered decalcification fixative

[0023] 1. Buffered decalcification solution Intended use: Used for fixation of fresh tissue samples. Only for in vitro analysis and detection purposes, not for therapeutic use. This product can be used for fixation of various types of bone tissue cells in the clinical human body. The fixation is uniform and stable, so that the tissue has a certain elasticity and is not easy to deform. It can well preserve the morphological structure of bone tissue cells and most of the substances in bone tissue cells, such as proteins, sugars, inorganic salts, pigments, etc., which is conducive to special staining, immunohistochemical staining and molecular biology testing. It is widely used in various types of hospitals and medical research and teaching units at all levels. The operation is simple and fast, and operators can master it without special training.

[0024] 2. Expected use environment of buffered decalcification solution: It can be used in various types of hospitals and medical research and teaching institutions at all levels. Working conditions: (1) Temperature: room temperature; (2) Humidity: not more than 85%; no special requirements for the environment.

[0025] 3. Applicable population: All kinds of people who need to undergo bone histology and cytology examinations, especially tissues suitable for immunohistochemical staining. It can well preserve the protein antigens of bone tissue cells, making the antigen localization clearer and meeting the quality control requirements of immunohistochemical technology.

[0026] 4. Product contraindications: None.

[0027] 5. Current application status of buffered decalcification solution: Tissue decalcification solution is a special tissue fixative and preservative used in medical laboratories for tissue specimen testing. It is widely used in hospital operating rooms, orthopedic laboratories, and pathology department tumor diagnosis and other laboratory applications. It is a standardized and normalized commonly used bone tissue decalcification solution. It is easy to use, safe and labor-saving, and is beneficial to the physical and mental health of operators and environmental protection. Pathological diagnosis is the gold standard for medical diagnosis. Standardized pathological diagnosis must first start with the fixation and decalcification of specimen bone tissue cells. The selection of standard fixation and decalcification solution is the key to fixation of specimens.

[0028] 6. Comparison of clinical usage of similar products already on the market: Similar products.

[0029] Testing method of the buffered decalcified fixative

[0030] The container should be large enough, preferably a wide-mouth bottle. Put the fixative first and then the bone specimen to prevent the tissue from sticking to the bottom of the container. The amount of fixative used: It is recommended to be 5 times the volume of the specimen.

[0031] Interpretation of Test Results

[0032] The surface and cross section of the tissue are grayish white without any blood color; the tissue is elastic and has a hard texture.

[0033] Limitations of the test method

[0034] This fixative is only used for fixation of ex vivo human bone tissue and animal bone tissue.

[0035] Product performance indicators

[0036] 1 Appearance and properties

[0037] 1.1 Transparent liquid.

[0038] 1.2 The packaging label is clear and accurate, the packaging is complete, and there is no liquid leakage.

[0039] 2Net content

[0040] Allowable range of net liquid content per bottle: 20±10.0%

[0041] 3. Accuracy

[0042] The relative deviation should be within ±10.0%

[0043] 4Minimum detection limit

[0044] Should not be greater than 0.5ng / ml

[0045] 5 Linear

[0046] In the range of 1.0ng / ml-1500ng / ml, the correlation coefficient (r) should not be less than 0.9900.

[0047] 6. Repeatability

[0048] The coefficient of variation CV should be ≤10%

[0049] 7Batch Difference

[0050] The coefficient of variation CV should be ≤10%

[0051] 8 Thermal stability

[0052] Within the validity period, the reagent should be placed at 37℃ for 3 days and its accuracy, minimum detection limit, linearity and repeatability should be tested. It should meet the requirements of 3, 4, 5 and 6.

[0053]

Notes

[0054] 1. If the laboratory receives insufficient decalcification solution in the tissue specimen, it should be immediately replenished with decalcification solution for fixation.

[0055] 2. Cover the bottle cap tightly to prevent the liquid from evaporating and send it to the laboratory for further processing.

[0056] 3. It should be used under well-ventilated conditions.

[0057] If it accidentally comes into contact with your skin or face, please rinse immediately with clean water or go to hospital for treatment.

[0058] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered exemplary and non-restrictive in all respects, and the scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes falling within the meaning and scope of the equivalent elements of the claims be included in the present invention.

[0059] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A buffered decalcification fixative, characterized in that: It is made by mixing together a variety of acidic substances, including lemon tree water solution, ammonium citrate, zinc sulfate and chloroform.

2. A buffered decalcification fixative according to claim 1, characterized in that: The mass ratio of the lemon tree aqueous solution, ammonium citrate, zinc sulfate and chloroform is 5:95:0.2:

3.

3. A buffered decalcification fixative according to claim 2, characterized in that: The concentration of the lemon tree aqueous solution per 5 ml is 7%.

4. A buffered decalcification fixative according to claim 2, characterized in that: The concentration of the ammonium citrate per 95 ml is 7.54%.

5. A buffered decalcification fixative according to claim 2, characterized in that: The concentration of the zinc sulfate per 0.2 ml is 1%.

6. Use of a buffered decalcification fixative according to any one of claims 1 to 5, characterized in that: Mainly used for fixation of fresh bone tissue samples.