Rich double-staining kit and use method thereof

A technology of dyeing reagents and kits, which is applied in the preparation of test samples, biological testing, sampling, etc., and can solve the problem of the difficulty in accurately distinguishing the positional relationship between tumor cells and elastic fibers, the difficulty in interpreting the pleura of lung cancer, and the difficulty in accurately displaying the relationship between tumor cells and the pleura. Elastic fiber layer relationship and other issues

Pending Publication Date: 2021-02-02
项征
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In 2000, the Japanese Lung Cancer Society basically agreed with the Hammar classification method (except Px), but there were differences in the division of T stages for tumors with visceral pleural invasion
The disadvantage is that the elastic fibers are black, which is the same color as the tumor cells, and are often mixed with pleural carbon foam. It is difficult to accurately distinguish the positional relationship between the tumor cells and the elastic fibers, making it difficult to interpret pleural invasion, and the subjective opinions of doctors are at odds.
In addition, immunohistochemical staining can be used. However, although immunohistochemical staining alone can display tumor cells through the reaction of antigen-antibody complexes, it cannot display elastic fibers, and it is also difficult to accurately display the relationship between tumor cells and the elastic fiber layer of the pleura. It is difficult to interpret the pleural invasion of lung cancer, so it is difficult to accurately stage, which in turn affects the choice of the next treatment plan for patients after surgery

Method used

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  • Rich double-staining kit and use method thereof
  • Rich double-staining kit and use method thereof
  • Rich double-staining kit and use method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 Ordinary elastic fiber dyeing method

[0068] In this embodiment, the elastic fiber dyeing kit purchased from Yili Company was used, referring to the instruction manual, and carried out according to the following operations:

[0069] 1) Paraffin tissue with conventional standardized treatment, sliced ​​at 3-5 μm;

[0070] 2) Bake in a 70°C incubator for 2 hours;

[0071] 3) Conventional xylene dewaxing twice, 10min each time, and then hydrating with gradient ethanol for a few seconds with 100% ethanol twice, 95% ethanol aqueous solution twice, 85% ethanol aqueous solution once, fully rinsed with tap water, and rinsed with distilled water 3 times;

[0072] 4) Add iron iodine hematoxylin for 10-15 minutes, wash with water;

[0073] 5) Differentiate the differentiation solution for a few seconds until the excess liquid fades away;

[0074] 6) Wash with water, add 95% ethanol to remove iodine for 1min;

[0075] 7) Counterstain in VG solution for 1 min;

[0...

Embodiment 2

[0080] Example 2 Double staining method using anti-TTF-1 antibody as primary antibody of the present invention

[0081] For paraffin tissue with routine standardized treatment, select the tissue block that needs to be stained, slice 3-5 μm, and then bake in a 70°C incubator for 2 hours; conventional xylene dewaxing 2 times, 10 minutes each time; then 100% ethanol 2 times each time 3 minutes each time, 95% ethanol water solution twice for 3 minutes each time, 85% ethanol water solution once for 3 minutes each time, followed by gradient ethanol hydration for 10 seconds, fully rinsed with tap water, rinsed with distilled water for 3 times; put the slices into pH 9.0, 1mM EDTA antigen retrieval solution Heated in a pressure cooker to 100°C for 1-5 minutes (2-3 minutes after gassing), cooled naturally to room temperature, washed 3 times with distilled water, 3 minutes each time; then sliced ​​in 3% hydrogen peroxide solution at room temperature Incubate for 10 minutes to remove e...

Embodiment 3

[0085] Example 3 Double staining method using AE1 / AE3 as the primary antibody of the present invention

[0086] For routine normalized paraffin tissue, select the tissue block that needs to be stained, slice 3-5 μm, and then bake in a 70°C incubator for 2 hours; conventional xylene dewaxing twice, each 10min; and then in 100% ethanol twice 3 minutes each time, 95% ethanol water solution twice for 3 minutes each time, 85% ethanol water solution once for 3 minutes each time, followed by gradient ethanol hydration for 10 seconds, fully rinsed with tap water, rinsed with distilled water for 3 times; put the slices into pH 9.0, 1mM EDTA antigen retrieval solution Heat it in a pressure cooker to 100°C for 1-5min (continue 2-3min after gassing), cool naturally to room temperature, wash 3 times with distilled water, 3min each time; then place the slices in 3% aqueous hydrogen peroxide solution, Incubate at room temperature for 10 minutes to remove endogenous peroxidase, then wash wi...

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Abstract

The invention provides a staining kit for all pathological tissue sections (especially lung cancer TNM staging) and a use method thereof. According to the staining kit and the use method thereof, immunohistochemical staining and special staining are combined for application, two staining modes can be completed on the same slice, staining time is shortened, displayed information amount is far larger than signals and information amount provided by traditional separate single staining, missed diagnosis of tiny infiltration is avoided, effective help can be provided for clinical later treatment, an important diagnosis basis is provided for defining clinical stages of a patient, and whether vascular tumor embolism exists in other types of tumors or not can be accurately judged. If other types of markers are selected, the markers can also be used as an index for cardiovascular disease discrimination.

Description

technical field [0001] The invention belongs to a pathological staining method in the technical field of biological detection. Background technique [0002] The morbidity and mortality of malignant tumors are increasing year by year, seriously threatening the health of patients. Accurate tumor staging is a prerequisite for precise treatment. For example, whether lung cancer has been invaded by the pleura is one of the key indicators for tumor staging and postoperative adjuvant chemotherapy. [0003] Regarding the relationship between pleural invasion of lung cancer and prognosis and treatment, as early as 1958, some scholars observed that pleural invasion of lung cancer was a poor prognostic factor. In 1975, visceral pleural invasion was used as TNM staging (T: primary tumor; N: regional Lymph nodes; M: distant metastasis) is a separate concept. In 1988, Hammar first proposed a grading method for pleural invasion of lung cancer: P0-no pleural invasion, the tumor did not ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/532G01N33/577G01N33/68G01N1/30
CPCG01N33/532G01N33/577G01N33/68G01N1/30G01N2001/302
Inventor 项征
Owner 项征
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