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Same-section multi-target protein immunohistochemical or immunofluorescent labeling method

A technique of immunohistochemistry and immunofluorescence staining, applied in the field of multi-target protein immunohistochemistry or immunofluorescence staining on the same section, which can solve the problems of color distinction and technical operation difficulties.

Active Publication Date: 2013-05-15
FUZHOU MAIXIN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the above-mentioned multi-antigen labeling technology is difficult to realize by immunohistochemical technology. At present, two types of antibody labeling can be carried out on the same section, that is, labeling horseradish enzyme to produce brown and alkaline phosphatase to produce blue-purple. However, there are certain difficulties in the color distinction and technical operation of the two, and the application of two different enzyme-labeled antibodies for immunohistochemical staining is rarely used

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 (immunohistochemistry 2 target markers):

[0023] Fix the wax block containing liver tumor tissue on the tissue slicer, and slice the tissue. When the blade runs to the target tissue, start picking, and put a series of continuous slices together in 60% ethanol solution for 1 minute. Then put it in hot water at 60°C for display, and select two consecutive slices. When picking up slides, the back of the first slice should face up, and the face of the second slice should face up immediately after picking up the slice. , to ensure that the surfaces of the two tissue sections used for staining belong to the same section. Two slices of the same section are regarded as a pair, and the detection of different proteins to be tested is carried out separately.

[0024]A pair of paraffin sections were dewaxed, hydrated, rinsed with tap water, and then heated to boiling EDTA solution (pH 9.0) at 100°C for 20 minutes. Add 150 μL of peroxidase blocking agent, incubate at ro...

Embodiment 2

[0025] Example 2 (immunohistochemical 4 target markers):

[0026] Fix the wax block containing pancreatic tumor tissue on the tissue slicer, and slice the tissue. When the blade runs to the target tissue, start picking, and put a series of serial slices together in 40% ethanol solution for 5 minutes. Then put it in hot water at 40°C to develop the slides, and select two consecutive slices. When picking up slides, the back of the first slice should face up, and the face of the second slice should face up immediately after picking up the slice. , to ensure that the surfaces of the two tissue sections used for staining belong to the same section. Two slices of the same section are regarded as a pair, and the detection of different proteins to be tested is carried out separately.

[0027] A pair of paraffin sections were dewaxed, hydrated, rinsed with tap water, and then heated to boiling EDTA solution (pH 9.0) at 100°C for 20 minutes. Add 160 μL of peroxidase blocking agent, in...

Embodiment 3

[0028] Example 3 (immunofluorescence 4 target labeling):

[0029] Fix the wax block containing pancreatic tumor tissue on the tissue slicer, and slice the tissue. When the blade runs to the target tissue, start picking, and put a series of serial slices together in 40% ethanol solution for 1 minute. Then put it in hot water at 40°C to develop the slides, and select two consecutive slices. When picking up slides, the back of the first slice should face up, and the face of the second slice should face up immediately after picking up the slices. Ensure that the surfaces of the two tissue sections used for staining belong to the same section. Two slices of the same section are regarded as a pair, and the detection of different proteins to be tested is carried out separately.

[0030] A pair of paraffin sections were dewaxed, hydrated, rinsed with tap water, and then heated to boiling EDTA solution (pH 9.0) at 100°C for 20 minutes. Rinse with PBS (the ratio is the same as in Exam...

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PUM

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Abstract

The invention provides a same-section multi-target protein immunohistochemical or immunofluorescent labeling method. The method comprises the following steps of: unfolding slices in corresponding liquid in the process of slicing a tissue wax block, selecting two continuously connected slices in the process of catching a target slice, placing the back surface of the first slice upwards in the process of catching the slice by employing a glass slide, and placing the front surface upwards in the process of catching the second slice, so that the condition that the surfaces of the two tissue slice for staining belong to the same slicing surface is ensured; and respectively performing immunohistochemical or immunofluorescent staining on different target proteins for two slices from the same slicing surface, performing overlap comparison on the tissue staining result in a same area, so as to realize existential analysis of a same cell or tissue site multi-target protein. The effect of performing multi-target protein labeling on the same cell or tissue site to be detected is achieved.

Description

technical field [0001] The invention belongs to the field of pathology. Specifically, the invention relates to a method for immunohistochemistry or immunofluorescence staining and marking of multi-target proteins in the same section. Background technique [0002] Tumor markers are physiologically active substances that are closely related to the growth and metabolism of tumor cells produced by tumor cells during malignant transformation, generation, development, invasion, and metastasis. They are often only found in embryonic tissues, and rarely exist in normal adult tissues, but their content in adult tumor tissues greatly exceeds that in normal tissues. By detecting their presence and concentration in corresponding cells or tissues To determine the primary tumor of a metastatic tumor of unknown origin, diagnose and classify the tumor and its characteristics, monitor surgery, chemotherapy, and radiotherapy, and evaluate the prognosis and treatment effect of the patient base...

Claims

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Application Information

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IPC IPC(8): G01N1/28
Inventor 林齐心陈赞烽熊玉林王小亚
Owner FUZHOU MAIXIN BIOTECH CO LTD
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