Antigen repair liquid for frozen tissue slice

Abstract

The invention provides an antigen repair liquid for frozen tissue slices, and particularly relates to the technical field of immunohistochemical detection, the antigen repair liquid comprises the following components: an antigen repair agent with a molar concentration of 1-5 mM; the mass fraction (m / V) of the tissue penetrant I is 0.01%-0.05%; a tissue penetrant II with a volume fraction (V / V) of 0.1-0.5%; and a pH stabilizer. The method comprises the following steps: soaking a cut frozen tissue section in an antigen repair solution, repairing for 5-10 minutes at 15-25 DEG C, flushing away the antigen repair solution left on the tissue section by using a TBS buffer solution, completing repair, and carrying out IHC staining. The method can quickly complete repair to achieve the effect of exposing the epitope, is simple and convenient to use, and can improve the overall dyeing effect.

Description

Technical Field

[0001] The present invention belongs to the technical field of immunohistochemical detection, and particularly relates to an antigen repair solution for frozen tissue sections. Background Art

[0002] Immunohistochemistry, an important technique commonly used in clinical tumor diagnosis, can not only help determine the type, nature, and origin of tumors, but also be used for treatment guidance and prognosis judgment of lesions, enabling the diagnosis of many difficult tumors. Its principle is a common pathological test method that uses known antibodies to detect antigens in the tissue to be tested, and finally presents the results in the form of tissue coloring. For the tissue to be detected, it is usually fixed to better preserve the antigen. However, aldehyde reagents commonly used for fixing cells or tissues will cause cross-linking between proteins after fixation, resulting in insufficient exposure of antigen sites and weakened staining effects. Therefore, paraffin-embedded tissue sections usually need to be antigen repaired, and the detection time takes 3 - 4 hours. However, in the actual clinical application process, the results of immunohistochemical staining need to be feedback within 30 minutes to guide surgeons to retain normal tissues as much as possible while removing tumor tissues, avoid damaging the normal tissue and organ functions of patients, and improve the quality of life of patients.

[0003] In order to develop rapid immunohistochemical staining reagents, the current mainstream idea is to improve the sensitivity of enzyme-labeled antibodies, that is, through a material containing a large number of active sites such as amino groups or aldehyde groups as an intermediate, and then coupling enzymes and antibodies respectively to form antibody polymers. However, the reaction time of such antibody polymers is long, the preparation difficulty is high, and the raw materials are expensive, greatly increasing the cost of rapid immunohistochemical staining and forming a burden on patients.

[0004] Existing technologies generally do not perform repair treatment on frozen tissues. Because conventional antigen repair solutions are designed for antigen repair of paraffin tissue sections and need to be used under high-temperature conditions above 70°C, with a repair time of up to 20 - 40 minutes, they do not meet the usage requirements of rapid immunohistochemical staining reagents.

[0005] In view of the above deficiencies, the present invention specifically develops an antigen repair solution suitable for frozen tissue sections. Incubating at room temperature for 5 - 10 minutes can achieve the effect of exposing antigen epitopes, with simple operation and convenient use. Summary of the Invention

[0006] The purpose of the present invention is to provide an antigen repair solution for frozen tissue sections, which can quickly complete the repair to achieve the effect of exposing antigen epitopes, is easy to use, and can improve the overall staining effect.

[0007] The present invention provides the following technical solutions:

[0008] An antigen repair solution for frozen tissue sections, comprising the following components:

[0009] An antigen repair agent with a molar concentration of 1 mM to 5 mM;

[0010] A tissue penetrant I with a mass fraction (m / V) of (0.01% to 0.05%, preferably 0.02%);

[0011] A tissue penetrant II with a volume fraction (V / V) of (0.1% to 0.5%, preferably 0.5%);

[0012] A pH stabilizer and water, and the pH of the antigen repair solution is 7.4 - 7.6.

[0013] Preferably, the antigen repair agent is sodium EDTA.

[0014] Preferably, the tissue penetrant I is sodium dodecyl sulfate.

[0015] Preferably, the tissue penetrant II is Triton.

[0016] Preferably, the pH stabilizer has a pH of 7.4 - 7.6.

[0017] Preferably, the pH stabilizer is a Tris-Hcl buffer solution with a molar concentration of 10 mM.

[0018] A method for repairing an antigen repair solution for frozen tissue sections is as follows: Immerse the cut frozen tissue sections in the antigen repair solution, and under the condition of 15 - 25 °C, repair for 5 - 10 min, then rinse the residual antigen repair solution on the tissue sections with TBS buffer solution, and the repair is completed, and the IHC staining step is carried out.

[0019] Advantages of the present invention:

[0020] 1. The present invention can act at room temperature, effectively remove the cross-linking between proteins, achieve the effect of more fully exposing antigen epitopes, quickly perform protein repair, is simple to operate and convenient to use, and can improve the staining effect of frozen tissue sections, effectively assisting in the clinical diagnosis of tumors.

[0021] 2. The present invention is simple to use, convenient to operate, and has a low configuration cost, laying a foundation for further and extensive clinical applications. Description of the Drawings

[0022] The drawings are used to provide a further understanding of the present invention, and constitute a part of the specification, and are used to explain the present invention together with the embodiments of the present invention, and do not constitute a limitation to the present invention. In the drawings:

[0023] Figure 1Staining diagram after repair with the antigen repair solution prepared in Example 1;

[0024] Figure 2 Staining diagram after repair with the antigen repair solution prepared in Example 2;

[0025] Figure 3 Staining diagram after repair with the antigen repair solution prepared in Example 3;

[0026] Figure 4 Staining diagram after repair without using the antigen repair solution of the present invention. Detailed implementation manners

[0027] Example 1

[0028] An antigen repair solution for frozen tissue sections, comprising disodium EDTA with a molar concentration of 1 mM, sodium dodecyl sulfate with a mass fraction (m / V) of 0.01%, Triton with a volume fraction (V / V) of 0.1%, Tris-Hcl buffer with a molar concentration of 10 mM, and water. The pH of the antigen repair solution is 7.4.

[0029] Repair method: Immerse the cut frozen tissue sections in this antigen solution, and repair at 15°C for 7 min. Further, rinse the residual repair solution on the tissue sections with TBS buffer, and perform IHC staining;

[0030] Example 2

[0031] An antigen repair solution for frozen tissue sections, comprising disodium EDTA with a molar concentration of 5 mM, sodium dodecyl sulfate with a mass fraction (m / V) of 0.05%, Triton with a volume fraction (V / V) of 0.3%, Tris-Hcl buffer with a molar concentration of 10 mM, and water. The pH of the antigen repair solution is 7.6;

[0032] Repair method: Immerse the cut frozen tissue sections in this antigen solution, and repair at 25°C for 10 min. Further, rinse the residual repair solution on the tissue sections with TBS buffer, and perform IHC staining.

[0033] Example 3

[0034] An antigen repair solution for frozen tissue sections, comprising guanidine isothiocyanate with a molar concentration of 2 mM, sodium dodecyl sulfate with a mass fraction (m / V) of 0.02%, Triton with a volume fraction (V / V) of 0.5%, Tris-Hcl buffer with a molar concentration of 10 mM, and water. The pH of the antigen repair solution is 7.5.

[0035] Repair method: Immerse the cut frozen tissue sections in this antigen solution, and repair at 20 °C for 5 min. Further, rinse the residual repair solution on the tissue sections with TBS buffer, and perform IHC staining.

[0036] Experimental control:

[0037] After repairing the frozen tissue sections with the antigen repair solutions obtained in Examples 1, 2, and 3, perform staining. The specific results are shown in Figure 1 、 Figure 2 and Figure 3 ;

[0038] For the frozen tissue sections, directly perform normal IHC staining without using the antigen repair solution of the present invention. The specific results are shown in Figure 4 ;

[0039] By Figure 1 and Figure 2 , Figure 3 and Figure 4 By comparison, it can be seen that at room temperature, repairing the frozen tissue sections with the antigen repair solution of the present invention for 5 - 10 minutes can improve the staining intensity and positive rate, and the overall staining effect is improved, especially in the case of poor staining effect.

[0040] The above are only the preferred embodiments of the present invention and are not used to limit the present invention. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements for some of the technical features. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. An antigen repair solution for frozen tissue sections, characterized in that: The components include: Antigen retrieval agent with a molar concentration of 1mM to 5mM; Tissue penetrant I with a mass fraction (m / V) of 0.01% to 0.05%; Tissue penetrant II with a volume fraction (V / V) of 0.1% to 0.5%; pH stabilizer and water, and the pH of the antigen retrieval solution is 7.4-7.6; The calculation of molar concentration, mass fraction and volume fraction is based on the entire antigen retrieval solution.

2. The antigen repair solution for frozen tissue sections according to claim 1, characterized in that: The antigen repair agent is EDTA sodium salt or guanidine isothiocyanate.

3. The antigen repair solution for frozen tissue sections according to claim 1, characterized in that: The molar concentration of the antigen repair agent is 2 mM; the mass fraction of the tissue penetrant I is 0.02%; the volume fraction of the tissue penetrant II is 0.5%; and the pH of the antigen repair solution is 7.

5.

4. The antigen repair solution for frozen tissue sections according to claim 3, characterized in that: Tissue penetrant I is sodium dodecyl sulfate.

5. The antigen repair solution for frozen tissue sections according to claim 3, characterized in that: The tissue penetrant II is Triton.

6. The antigen repair solution for frozen tissue sections according to claim 1, characterized in that: The pH stabilizer is pH 7.4-7.

6.

7. The antigen repair solution for frozen tissue sections according to claim 6, characterized in that: The pH stabilizer was a Tris-HCl buffer with a molar concentration of 10 mM.

8. The antigen repair solution for frozen tissue sections according to claim 1, wherein the repair method is as follows: Immerse the cut frozen tissue sections in antigen repair solution at 15-25°C for 5-10 minutes, then use TBS buffer to rinse off the residual antigen repair solution on the tissue sections. The repair is completed, and then IHC staining is performed.

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