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Novel tissue slice immunohistochemical operation method

A technology of tissue slicing and operation method, which is applied in the field of laboratory inspection and analysis, can solve the problems of low work efficiency and inaccurate dripping results, etc., and achieve the effects of convenient operation, conducive to standardized operation of experiments, and simple methods

Active Publication Date: 2016-10-12
SICHUAN KINGMED DIAGNOSTICS CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A new immunohistochemical operation method for tissue sections is now provided. By adjusting the operation steps of the existing operation method, the technical problems such as low work efficiency and inaccurate dropping results when adding antibodies in immunohistochemical operations are solved.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A novel immunohistochemical operation method for tissue sections, comprising the following steps: 1) blocking endogenous catalase: soaking in methanol with 0.3% hydrogen peroxide for 30 minutes;

[0032] 2) Staining: staining with hematoxylin for 2 minutes;

[0033] 3) Add I antibody dropwise: add I antibody 50ul dropwise, let stand at room temperature for 40 minutes;

[0034] 4) Add II antibody dropwise: add II antibody 40-50ul dropwise, and let it stand at room temperature.

[0035] Other routine experimental procedures such as slicing, copying, dewaxing, hydration, antigen retrieval, color development, and sealing of tissue sections that are not involved in this example are common knowledge in the art and will not be repeated here.

Embodiment 2

[0037] A novel immunohistochemical operation method for tissue sections, including the following steps: 1) Slicing and copying: Bake the tissue sections at 60°C for 20 minutes;

[0038] 2) Dewaxing and hydration: Soak tissue sections in xylene for 10 minutes, replace xylene and then soak for 10 minutes, then soak in absolute ethanol, 95% ethanol, and 70% ethanol for 5 minutes respectively;

[0039] 3) Block endogenous catalase: soak in methanol with 0.3% hydrogen peroxide for 30 minutes, then wash with distilled water for 6 minutes, and rinse with PBS for 6 minutes;

[0040] 4) Staining: Stain with hematoxylin for 2 minutes and differentiate with hydrochloric acid alcohol;

[0041] 5) Add I antibody dropwise: add I antibody 50ul dropwise, let stand at room temperature for 1 hour or overnight at 4°C or 1 hour at 37°C;

[0042] 6) Add II antibody dropwise: add II antibody 50ul dropwise, 37°C for 1 hour;

[0043] 7) Add III antibody dropwise, the dropping temperature is 37°C, a...

Embodiment 3

[0045] A novel immunohistochemical operation method for tissue sections, including the following steps: 1) Slicing and copying: Bake the tissue sections at 60°C for 20 minutes;

[0046] 2) Dewaxing and hydration: Soak tissue sections in xylene for 10 minutes, replace xylene and then soak for 10 minutes, then soak in absolute ethanol, 95% ethanol, and 70% ethanol for 5 minutes respectively;

[0047] 3) Block endogenous catalase: soak in methanol with 0.3% hydrogen peroxide for 30 minutes, then wash with distilled water for 6 minutes, and rinse with PBS for 6 minutes;

[0048] 4) Staining: Stain with hematoxylin for 2 minutes and differentiate with hydrochloric acid alcohol;

[0049] 5) Add I antibody dropwise: add I antibody 50ul dropwise, let stand at room temperature for 1 hour or overnight at 4°C or 1 hour at 37°C;

[0050] 6) Add II antibody dropwise: add II antibody 50ul dropwise, 37°C for 1 hour;

[0051] 7) Rinse the tissue sections with PBS buffer for 3 times in total...

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Abstract

The invention discloses a novel tissue slice immunohistochemical operation method. The novel tissue slice immunohistochemical operation method includes the steps of 1), closing endogenous catalase and soaking by 0.3% methanol-hydrogen peroxide for 30 minutes; 2), staining by hematoxylin for 2 minutes; 3), dropwise adding 40 microlitres of antibody I and standing at room temperature for 40 minutes-1 hour; 4), dropwise adding 40-50 microlitres of antibody II and standing at room temperature or 37 DEG C for 1 hour. The novel tissue slice immunohistochemical operation method has the advantages that a conventional operation step that the antibody I and the antibody II are stained by the hematoxylin before being dropwise added is changed to be beneficial to developing of the antibody I and the antibody II which are dropwise added. The novel tissue slice immunohistochemical operation method is simple in operation step and succeeds in changing existing operation effects only by making changes on operation sequences of existing operation steps, adopted reagents and operation time, and the antibodies are added dropwise after staining to facilitate confirmation of sizes and positions of tissues.

Description

technical field [0001] The invention relates to a laboratory test and analysis method, in particular to a novel immunohistochemical operation method for tissue sections. Background technique [0002] Immunohistochemistry is the application of the principle of immunological antigen-antibody reaction, that is, the principle of specific reaction between antigen and antibody. Through chemical reaction, the chromogenic agent of the labeled antibody, such as fluorescein, enzyme, metal ion, isotope, etc., is developed to determine the reaction tissue cells. Antigen, its localization, qualitative and quantitative research. [0003] For immunohistochemical operations, experimenters often encounter tissue sections that have no color after repairing. When adding I antibody and II antibody, they do not know the specific dripping location and tissue, so they can only take a large area. Dropping, resulting in inaccurate dropping results, unable to locate antigens in tissue cells, and una...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53
CPCG01N33/5302
Inventor 欧阳小峰刘叶谷和林李军秀
Owner SICHUAN KINGMED DIAGNOSTICS CENT
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