A tissue antigen retrieval solution

Abstract

The application discloses a kind of tissue antigen repair liquids, it is related to biomedical technology field, and its technical points are: the tissue antigen repair liquid of the application is by composition 0.125% trypsin, calcium magnesium pbs solution without;0.125% trypsin is dissolved in dilution one times calcium magnesium pbs without, and ph is adjusted to 7.0, i.e. the tissue antigen repair liquid of the application can be prepared.This tissue liquid can significantly improve the effect when paraffin section carries out antigen repair processing, so that slice sample presents higher image definition and more detailed tissue structure performance under microscopic observation.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a tissue antigen repair solution. Background Technology

[0002] Tissue antigen retrieval solutions are widely used reagents in biomedical research, primarily to improve the detectability of antigens in tissue sections. They are commonly used in immunohistochemistry (IHC) and fluorescence histochemistry experiments. Their main function is to restore antigens that may have been damaged or masked during tissue fixation, optimizing antibody binding ability and thus improving the sensitivity and specificity of experiments. Tissue antigen retrieval solutions typically contain buffer salts (such as phosphate or Tris buffer), enzymes, and additives to help remove substances that mask antigens. In use, tissue sections must first be dewaxed and hydrated, then immersed in the retrieval solution, appropriately treated, washed to remove residues, and finally stained with antibodies. This solution is of great significance in cancer research, histopathology, and basic biological research, effectively improving the accuracy and reliability of experimental results.

[0003] However, frozen sections are usually thicker, making it difficult to obtain a clearer visual effect. The paraffin section heat restoration method has certain drawbacks, such as poor safety when heated to 95 degrees Celsius, and the operation is time-consuming, laborious, and inconvenient.

[0004] Therefore, the present invention aims to provide a tissue antigen repair solution to solve the above-mentioned problems. Summary of the Invention

[0005] The purpose of this invention is to solve the above-mentioned problems and provide a tissue antigen repair solution.

[0006] To achieve the above objectives, the technical solution of the present invention is as follows:

[0007] This invention provides a tissue antigen repair solution, which consists of 0.125% trypsin and calcium- and magnesium-free PBS solution. The tissue antigen repair solution is prepared by dissolving 0.125% trypsin in calcium- and magnesium-free PBS solution diluted by one-fold and adjusting the pH to 7.0.

[0008] The preparation method of calcium- and magnesium-free PBS is as follows: Weigh 8.0g NaCl, 0.2g KCl, 1.44g Na2HPO4 and 0.24g KH2PO4 and dissolve them in 800mL distilled water. Adjust the solution to 7.4 with HCl and finally add distilled water to make up to 1L.

[0009] Compared with existing technologies, the beneficial effects of this solution are:

[0010] The tissue fluid of this invention can significantly improve the effect of antigen retrieval treatment on paraffin sections, resulting in higher image clarity and more detailed tissue structure under microscopic observation. Compared with the traditional 95°C high-temperature heat retrieval method, this tissue fluid treatment method can effectively avoid tissue sample detachment during the retrieval process. At the same time, the entire operation does not require high-temperature heating, which not only significantly improves experimental safety but also greatly simplifies the operation process, saving more time and effort. Attached Figure Description

[0011] Figure 1 This is a comparative schematic diagram of tissue antigen repair solutions in the embodiments of the present invention, wherein a is the antigen repair solution of this application; b is 95° antigen repair; and c is a commercially available antigen repair solution.

[0012] Figure 2 This is a schematic diagram illustrating the effect of frozen sections in an embodiment of the present invention;

[0013] Figure 3 This is a schematic diagram of the effect of the repair solution on the antigen repair of a mouse paraffin fracture sample in an embodiment of the present invention, where green represents CTSK, red represents trem2, and blue represents trap;

[0014] Figure 4 This is a schematic diagram of the antigen repair effect of the repair solution on a rat paraffin femur sample in an embodiment of the present invention, where white represents OSX, red represents CD31, and blue represents DAPI;

[0015] Figure 5 This is a schematic diagram of the antigen repair effect of the repair solution on a rat paraffin femur sample in an embodiment of the present invention, where white represents COL-1, red represents VEGFA, and blue represents dapi;

[0016] Figure 6 This is an immunofluorescence retrieval effect image of paraffin-embedded brain sections in an embodiment of the present invention, where the red area represents ctsh;

[0017] Figure 7 This is an immunofluorescence image of the repair effect of paraffin-embedded muscle sections in an embodiment of the present invention, where the yellow area represents laminin. Detailed Implementation

[0018] To enable those skilled in the art to better understand the present invention, the technical solution of the present invention will be described in further detail below with reference to the embodiments and accompanying drawings. Obviously, the described embodiments are merely some, not all, of the embodiments of the present invention. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort should fall within the scope of protection of the present invention.

[0019] It should be noted that, unless otherwise specified, the embodiments and features described in the present invention can be combined with each other. The present invention will now be described in detail with reference to the embodiments.

[0020] Example:

[0021] This invention provides a tissue antigen repair solution, which consists of 0.125% trypsin and calcium- and magnesium-free PBS solution. The tissue antigen repair solution is prepared by dissolving 0.125% trypsin in calcium- and magnesium-free PBS solution diluted by one-fold and adjusting the pH to 7.0.

[0022] The preparation method of calcium- and magnesium-free PBS is as follows: Weigh 8.0g NaCl, 0.2g KCl, 1.44g Na2HPO4 and 0.24g KH2PO4 and dissolve them in 800mL distilled water. Adjust the solution to 7.4 with HCl and finally add distilled water to make up to 1L.

[0023] Experiments were conducted to observe the effects of four different treatment methods on the expression rates of OPN and OSX antibodies. The results showed that using the tissue antigen retrieval solution of this invention, the expression rates of both OPN and OSX antibodies reached 100%, demonstrating the best performance. After antigen retrieval treatment at 95°C, the expression rates of OPN and OSX antibodies were 85% and 80%, respectively, both maintaining high expression levels. However, when using commercially available antigen retrieval reagents, the expression rates of both OPN and OSX antibodies were 0%, making them completely undetectable. In the frozen section method, the expression rate of OPN antibody was only 50%, but the expression rate of OSX antibody was as high as 95%, indicating a significant difference in the applicability of this method to the two antibodies. In conclusion, the tissue antigen retrieval solution of this invention can simultaneously achieve complete expression of both OPN and OSX antibodies, significantly superior to the other three conventional treatment methods.

[0024] The above specific embodiments are merely explanations of the present invention and are not intended to limit the present invention. After reading this specification, those skilled in the art can make modifications to these embodiments without contributing any inventive step, but as long as they are within the scope of the claims of the present invention, they are protected by patent law.

Claims

1. A tissue antigen repair solution, characterized in that: The tissue antigen repair solution consists of 0.125% trypsin and calcium- and magnesium-free PBS solution.

2. The tissue antigen repair solution as described in claim 1, characterized in that: The preparation method of the tissue antigen repair solution is as follows: dissolve trypsin with a mass fraction of 0.125% in calcium- and magnesium-free PBS diluted by one time, and adjust the pH to 7.

0.

3. The tissue antigen repair solution as described in claim 4, characterized in that: The preparation method of the calcium- and magnesium-free PBS is as follows: Weigh 8.0g NaCl, 0.2g KCl, 1.44g Na2HPO4 and 0.24g KH2PO4 and dissolve them in 800mL of distilled water. Adjust the solution to 7.4 with HCl and finally add distilled water to make up to 1L.

4. The tissue antigen repair solution as described in claim 1, characterized in that: The preparation method of the tissue antigen repair solution is as follows: The tissue antigen repair solution can significantly improve the effect of antigen repair treatment on paraffin sections, so that the section samples exhibit higher image clarity and more detailed tissue structure under microscopic observation.

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