Histological classification immunohistochemical multiple staining detection method for lung cancer

A detection method, immunohistochemical technology, applied in the field of lung cancer histological typing immunohistochemical multiple staining detection, can solve the problems of time-consuming and labor-intensive detection of immunohistochemical technology, difficult typing and other problems, achieve intuitive comparison of experimental results, staining The effect of fewer steps and shorter experiment time

Active Publication Date: 2014-12-17
FUZHOU MAIXIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the disadvantages of difficult typing of lung cancer histology and the time-consuming and laborious detection of immunohistochemical techniques respectively, and provide a method for detecting lung cancer histological typing and immunohistochemical multiple staining

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  • Histological classification immunohistochemical multiple staining detection method for lung cancer
  • Histological classification immunohistochemical multiple staining detection method for lung cancer
  • Histological classification immunohistochemical multiple staining detection method for lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The research objects were formaldehyde-fixed-paraffin-embedded lung adenocarcinoma tissues (Department of Pathology, Fujian Provincial Hospital). The steps of immunohistochemical experiment are as follows:

[0028] (1) The tissue slices were dry-baked in a 67°C incubator for 2 hours.

[0029] (2) Conventional xylene dewaxing 3 times, 6 minutes each time, hydration in 100%, 100%, 95%, 85% gradient ethanol, 3 minutes each time, and finally rinsed with tap water.

[0030] (3) Boil and repair directly in EDTA antigen retrieval solution with pH 9.0, cool naturally to room temperature, rinse with tap water, and rinse with PBS for 3×3 minutes.

[0031] (4) Incubate in 3% hydrogen peroxide at room temperature for 10 minutes to block endogenous peroxidase, wash with PBS for 3×3 minutes.

[0032] (5) Block with normal animal serum for 10 minutes, shake off excess serum, do not wash, add Napsin A, TTF-1, Desmoglein3 mixed primary antibody dropwise, the antibody titers in the mix...

Embodiment 2

[0039] The research objects were formaldehyde-fixed-paraffin-embedded lung squamous cell carcinoma tissues (Department of Pathology, Fujian Provincial Hospital). The steps of immunohistochemical experiment are as follows:

[0040] (1) The tissue slices were dry-baked in a 67°C incubator for 2 hours.

[0041] (2) Conventional xylene dewaxing 3 times, 6 minutes each time, hydration in 100%, 100%, 95%, 85% gradient ethanol, 3 minutes each time, and finally rinsed with tap water.

[0042] (3) Boil and repair directly in EDTA antigen retrieval solution with pH 9.0, cool naturally to room temperature, rinse with tap water, and rinse with PBS for 3×3 minutes.

[0043] (4) Incubate in 3% hydrogen peroxide at room temperature for 10 minutes to block endogenous peroxidase, wash with PBS for 3×3 minutes.

[0044] (5) Block with normal animal serum for 10 minutes, shake off excess serum, do not wash, add Napsin A, TTF-1, CK5 / 6 mixed primary antibody dropwise, and the titers of each anti...

Embodiment 3

[0051] The research objects were formaldehyde-fixed-paraffin-embedded lung small cell carcinoma tissues (Department of Pathology, Fujian Provincial Hospital). The steps of immunohistochemical experiment are as follows:

[0052] (1) The tissue slices were dry-baked in a 67°C incubator for 2 hours.

[0053] (2) Conventional xylene dewaxing 3 times, 6 minutes each time, hydration in 100%, 100%, 95%, 85% gradient ethanol, 3 minutes each time, and finally rinsed with tap water.

[0054] (3) Boil and repair directly in EDTA antigen retrieval solution with pH 9.0, cool naturally to room temperature, rinse with tap water, and rinse with PBS for 3×3 minutes.

[0055] (4) Incubate in 3% hydrogen peroxide at room temperature for 10 minutes to block endogenous peroxidase, wash with PBS for 3×3 minutes.

[0056] (5) Block with normal animal serum for 10 minutes, shake off excess serum, do not wash, add Napsin A, TTF-1, p40, TRIM29 mixed primary antibody dropwise, and the titers of each a...

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Abstract

The invention belongs to the technical field of immunohistochemistry, and in particular relates to a histological classification immunohistochemical multiple staining detection method for lung cancer. The primary antibody in the detection method is various mixed primary antibodies. In order to overcome the defects that existing lunge cancer is not easy to classify in histology, and detection based on an immunohistochemical technology consumes time and wastes labor, the staining process is simple and quick by adopting the method, so that more abundant information with strong contrast can be obtained on a same slice quickly and conveniently. According to the invention, the mixed primary antibodies and various detection amplifying systems are applied to identifying histological classification of lung cancer, so that not only can the same sensitivity and specificity of each antibody in the mixed antibodies be maintained in comparison with those in simple staining, but also the method has the advantages of less staining steps, short experimental time, high stability, intuitionistic comparison of experimental result, far great information amount than single staining and the like, especially has remarkable advantages of biopsic pathological diagnosis with small specimen amount.

Description

technical field [0001] The invention belongs to the technical field of immunohistochemistry, in particular, the invention relates to a lung cancer histological typing immunohistochemical multiple staining detection method. Background technique [0002] Lung cancer is the most common malignant tumor of the respiratory system and the most common cause of death. It can be divided into small cell lung cancer and non-small cell lung cancer. The common histological types of the latter are squamous cell carcinoma, adenocarcinoma, and large cell carcinoma, and occasionally some other types or mixed cell types are less frequent. The distinction between lung small cell carcinoma and non-small cell carcinoma has always been considered the most important issue in pathological diagnosis, and the distinction between the two has important therapeutic significance. Pathologists do not make the differential diagnosis of the two based on cell morphology and immunohistochemical methods. not d...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574
Inventor 杨清海王小亚
Owner FUZHOU MAIXIN BIOTECH CO LTD
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