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Immunohistochemical staining kit containing pigment tissues and staining method

A technology of immunohistochemistry and staining reagents, applied in the field of immunostaining, which can solve the problems of affecting the interpretation of immunohistochemistry results, covering the morphology and structure of tissue cells, and low sensitivity of the detection system, so as to achieve clear and distinguishable immunohistochemistry results and staining background. Clean, signal-clear results

Active Publication Date: 2019-10-11
河南赛诺特生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A large number of these substances are distributed on the tissue cells, covering the morphological structure of the tissue cells. When the traditional immunohistochemical method is used, the tan or tan color of DAB is difficult to distinguish from the color of these substances, which seriously affects its immunity. Interpretation of histochemical results
[0004] There are generally two traditional solutions. One method is to remove the melanin from the tissue before the immunohistochemical test, and use a strong oxidant to remove the pigment. Partial antigen loss and tissue detachment during the test process. At the same time, other substances such as carbon particles cannot be removed by strong oxidants. This method has many limitations; The FAST RED detection system is used for the detection of this type of tissue. The positive particles of this color development system are red, which can be distinguished from pigments. However, this detection system has low sensitivity, long operation time, and easy to produce background coloring

Method used

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  • Immunohistochemical staining kit containing pigment tissues and staining method
  • Immunohistochemical staining kit containing pigment tissues and staining method
  • Immunohistochemical staining kit containing pigment tissues and staining method

Examples

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Embodiment 1

[0049] In this embodiment, the method for immunohistochemical staining of pigmented tissue comprises the following steps:

[0050] 1) Take the pigmented tissue section to be tested and add peroxidase blocking agent dropwise to block endogenous peroxidase;

[0051] 2) Incubate the tissue sections with the target antibody, and then incubate with the peroxidase-labeled secondary antibody polymer;

[0052] 3) Incubate and stain the tissue sections at room temperature for 5-10 minutes using the blue dye chromogenic solution; the blue dye chromogenic solution includes: the blue dye substrate based on the peroxidase system and the blue dye that matches the blue dye substrate buffer;

[0053] 4) Use the red contrast agent to stain the tissue sections.

Embodiment 2

[0055] In this embodiment, the method for immunohistochemical staining of pigmented tissue comprises the following steps:

[0056] 1) Take one melanoma tissue, prepare 6 paraffin tissue sections with a thickness of 3 microns, and bake the slices at 60°C for 1 hour.

[0057] 2) Routine dewaxing and hydration of tissue sections: incubate xylene Ⅰ and Ⅱ for 15 minutes each, ethanol Ⅰ and Ⅱ for 5 minutes each, 95% ethanol, 80% ethanol and 70% ethanol for 5 minutes each, Wash with pure water, soak for 3 minutes*3 times.

[0058] 3) Antigen restoration: use Tris-EDTA (pH9.0) restoration solution, repair under high pressure for 3 minutes, cool naturally, wash with PBST, soak for 3 minutes*3 times.

[0059] 4) Blocking with blocking agent: Incubate with peroxidase blocking agent to block endogenous peroxidase; 100 microliters of reagent per slice, incubate at room temperature for 5 minutes, wash with PBST buffer, soak for 3 minutes*3 times.

[0060] 5) Antibody incubation: Sections ...

Embodiment 3

[0068] In this embodiment, the method for immunohistochemical staining of pigmented tissue comprises the following steps:

[0069] 1) Take one melanoma tissue and one lung adenocarcinoma tissue, prepare two paraffin tissue sections with a thickness of 3 microns, and bake the slices at 60°C for 1 hour.

[0070] 2) Routine dewaxing and hydration of tissue sections: incubate xylene Ⅰ and Ⅱ for 15 minutes each, ethanol Ⅰ and Ⅱ for 5 minutes each, 95% ethanol, 80% ethanol and 70% ethanol for 5 minutes each, Wash with pure water, soak for 3 minutes*3 times.

[0071] 3) Antigen restoration: use Tris-EDTA (pH9.0) restoration solution, repair under high pressure for 3 minutes, cool naturally, wash with PBST, soak for 3 minutes*3 times.

[0072] 4) Blocking with blocking agent: Incubate with peroxidase blocking agent to block endogenous peroxidase; 100 microliters of reagent per slice, incubate at room temperature for 5 minutes, wash with PBST buffer, soak for 3 minutes*3 times.

[00...

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Abstract

The invention relates to an immunohistochemical staining kit containing pigment tissues and a staining method, belonging to the technical field of immunostaining. In the staining kit, a peroxidase block can block endogenous peroxidase of tissues, so as to prevent the endogenous peroxidase from affecting a staining result; a peroxidase-labeled secondary antibody polymer is based on a HRP enzyme labeling system and has high sensitivity; a blue stained substrate based on a peroxidase system and a blue stained buffer solution matching the blue stained substrate make up the blue stained color developing solution, under catalysis of the HRP enzyme, the blue stained color developing solution forms blue granular precipitates that can be observed under a microscope. The positive part of the tissuecan be positioned accurately, the signal is clear, the blue granular precipitates are in sharp contrast to color substances (melanin or carbon particles), and the immunohistochemical result is clear and distinctive. The enzyme labeling system in the invention has high sensitivity, high specificity, and has a clear staining background.

Description

technical field [0001] The invention relates to an immunohistochemical staining kit and a staining method for pigmented tissue, belonging to the technical field of immunostaining. Background technique [0002] Immunohistochemical staining technology is the application of the basic principle of immunology - antigen-antibody reaction, that is, the principle of specific combination of antigen and antibody, through chemical reaction to make the chromogen of the labeled antibody develop color to determine the antigen (polypeptide and protein) in the tissue section or cell. ), the technology for its positioning, qualitative or quantitative research has the characteristics of specificity, sensitivity and accuracy, and integrates form, function and metabolism. This technology plays an important role in modern pathological diagnosis. [0003] At present, the most conventional immunohistochemical chromogenic method is based on the HRP enzyme-labeled system, and the chromogenic substr...

Claims

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Application Information

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IPC IPC(8): G01N1/31G01N33/532
CPCG01N1/31G01N33/532G01N2001/302
Inventor 牛银银刘玲玲米贯勋杨潇燕臧素芳齐华李道明张会娟曹巍
Owner 河南赛诺特生物技术有限公司
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