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A kind of immunohistochemical operation method

An operation method and immunohistochemical technology, applied in the field of laboratory inspection and analysis, can solve the problems of low work efficiency and large waste of solvent, and achieve the effects of convenient operation, avoidance of waste and conducive to standardized experimental operation.

Active Publication Date: 2018-12-28
SICHUAN KINGMED DIAGNOSTICS CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

An immunohistochemical operation method is now provided. By adjusting the operation steps of the existing operation method, the technical problems such as low work efficiency and large waste of solvent when adding antibodies in the immunohistochemical operation are solved.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] An immunohistochemical operation method, comprising the following steps: 1) sectioning and copying: placing the tissue section at room temperature for 60 minutes;

[0035] 2) Dewaxing and hydration: soak the tissue sections in xylene for 10 minutes, replace the xylene and then soak for 10 minutes, and then soak in absolute ethanol, 95% ethanol, and 70% ethanol for 5 minutes respectively;

[0036] 3) Antigen retrieval: Heat the 0.01M sodium citrate buffer solution with a pH of 6.0 to 95°C in an electric furnace, put it into tissue slices and heat for 10 minutes;

[0037] 4) Block endogenous catalase: soak in 0.3% hydrogen peroxide in methanol for 30 minutes;

[0038] 5) Staining: staining with hematoxylin for 2 minutes;

[0039] 6) Add I antibody dropwise: add 50 microliters of I antibody dropwise, and let stand at room temperature for 1 hour;

[0040] 7) Add II antibody dropwise: add 40-50 microliters of II antibody dropwise, and let it stand at room temperature;

[...

Embodiment 2

[0044] An immunohistochemical operation method, comprising the following steps: 1) sectioning and copying: baking the tissue section at 60°C for 20 minutes;

[0045] 2) Dewaxing and hydration: soak the tissue sections in xylene for 10 minutes, replace the xylene and then soak for 10 minutes, and then soak in absolute ethanol, 95% ethanol, and 70% ethanol for 5 minutes respectively;

[0046] 3) Antigen retrieval: Heat pH6.0 0.01M sodium citrate buffer solution to 95°C in a water bath, put it into tissue slices and heat for 15 minutes;

[0047] 4) Block endogenous catalase: soak in 0.3% hydrogen peroxide in methanol for 30 minutes, then wash with distilled water for 6 minutes, and rinse with PBS for 6 minutes;

[0048] 5) Staining: Staining with hematoxylin for 2 minutes and differentiation with hydrochloric acid alcohol;

[0049] 6) Add I antibody dropwise: add 50 microliters of I antibody dropwise, let stand at room temperature for 1 hour or overnight at 4°C or 1 hour at 37°C...

Embodiment 3

[0055] An immunohistochemical operation method, comprising the following steps:

[0056] 1) Slicing and copying: place the tissue slices at room temperature for 60 minutes;

[0057] 2) Dewaxing and hydration: soak the tissue sections in xylene for 10 minutes, replace the xylene and then soak for 10 minutes, and then soak in absolute ethanol, 95% ethanol, and 70% ethanol for 5 minutes respectively;

[0058] 3) Antigen retrieval: Heat 0.01M sodium citrate buffer solution with a pH of 6.0 to 95°C in a water bath, put it into tissue slices and heat for 12 minutes;

[0059] 4) Block endogenous catalase: soak in 0.3% hydrogen peroxide in methanol for 30 minutes;

[0060] 5) Staining: staining with hematoxylin for 2 minutes;

[0061] 6) Add I antibody dropwise: add 50 microliters of I antibody dropwise, and let it stand at 37°C for 1 hour; wherein, the composition of I antibody includes 3% BSA and 10% serum

[0062] 7) Add II antibody dropwise: add 40-50 microliters of II antibody...

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Abstract

The invention discloses an immunohistochemical operation method. The immunohistochemical operation method comprises the following steps of: (1) slicing and baking slices; (2) dewaxing and hydrating; (3) repairing an antigen; (4) sealing endogenous catalase; (5) dyeing; (6) dropwise adding an antibody I; (7) dropwise adding an antibody II; (8) developing, namely developing by adopting a DAB developing agent at a room temperature; and (9) sealing the slices. According to the technical scheme disclosed by the invention, a conventional operation step of dropwise adding the antibody I and the antibody II before dyeing by haematoxylin, the antibody can be easily developed after the antibody I and the antibody II are dropwise added. The immunohistochemical operation method is simple in operation steps; only an operation sequence of existing operation steps, adopted reagents and operation time are changed and the change of an existing operation effect can be realized; the antibodies are dropped after dyeing so that the size and position of tissues can be conveniently grasped. The operation steps are simple and certain technical detail problems in experiment check analysis can be effectively solved; standard operation of an experiment are easy to realize, wastes of experiment reagents can be avoided and certain errors in the experiment are also reduced as much as possible.

Description

technical field [0001] The invention relates to a laboratory test and analysis method, in particular to an immunohistochemical operation method. Background technique [0002] When the laboratory was doing immunohistochemical antibody titration, it was found that the incubation box and buffer were colorless, and the tissue sections had no color after repairing. The antibody had no sense of direction, and the size and position of the antibody tissue could not be grasped. You can only keep looking carefully and increase the amount of dripping. [0003] During the whole process of antibody dripping, multiple drops, wrong drops, and missing drops will cause waste of reagents, increase testing costs, affect results, and delay film production time. [0004] In the process of film production, it is necessary to search carefully and drop more reagents, which will increase the working steps, prolong the working time, reduce the working efficiency, and indirectly increase the labor co...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N1/30G01N1/44G01N21/78
CPCG01N1/30G01N1/44G01N21/78
Inventor 欧阳小峰刘叶谷和林李军秀
Owner SICHUAN KINGMED DIAGNOSTICS CENT
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