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Pretreatment liquid and pretreatment method used for fluorescence in situ hybridization of FFPE tissue slices

A technology of fluorescence in situ hybridization and pretreatment solution, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc., can solve problems affecting the interpretation of results, simplify the dewaxing step, reduce autofluorescence and background, and save the effect of time

Inactive Publication Date: 2016-08-17
WUHAN HEALTHCHART BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

When performing fluorescence in situ hybridization on certain tissue cells with vigorous metabolism or tissues with a large amount of collagen and elastin or reticular fibers, the hybridization results show strong autofluorescence and background under a fluorescent microscope, which seriously affects the interpretation of results , and the traditional processing method can not solve this problem very well

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  • Pretreatment liquid and pretreatment method used for fluorescence in situ hybridization of FFPE tissue slices
  • Pretreatment liquid and pretreatment method used for fluorescence in situ hybridization of FFPE tissue slices
  • Pretreatment liquid and pretreatment method used for fluorescence in situ hybridization of FFPE tissue slices

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Embodiment 1

[0034] In this embodiment, the breast cancer FFPE tissue sample is selected as a test sample, and the pretreatment method of the present invention is used for pretreatment, including the following steps:

[0035] (1) Baking slices: place the tissue slices on a 65°C grilling machine and bake the slices for 3-5 minutes;

[0036] (2) Pretreatment solution I treatment: immerse the tissue section in pretreatment solution I preheated to 68°C for 5 minutes; the components of the pretreatment solution I are 1wt% ethylphenyl polyethylene glycol and 0.5wt% alkylphenol polyoxyethylene ether.

[0037] (3) Pretreatment solution II treatment: take out the tissue sections in the pretreatment solution I, drain the remaining pretreatment solution I on absorbent paper, and then immerse the tissue sections in the pretreatment solution II preheated to 72°C for 10 min, Shake the slide up and down once every 2-3 minutes; the pretreatment solution II consists of 0.5 wt% ethylphenyl polyethylene gly...

Embodiment 2

[0073] In this embodiment, a lung adenocarcinoma FFPE tissue sample is selected as a test sample, and the pretreatment method for fluorescence in situ hybridization FFPE tissue section of the present invention is used for pretreatment, including the following steps:

[0074] (1) Baking slices: place the tissue slices on a 65°C grilling machine and bake the slices for 3-5 minutes;

[0075] (2) Pretreatment solution I treatment: immerse the tissue section in pretreatment solution I preheated to 68°C for 5 minutes; the pretreatment solution I is 1 wt% ethylphenyl polyethylene glycol and 0.5 wt% alkylphenol poly Oxyethylene ether.

[0076] (3) Pretreatment solution II treatment: take out the tissue sections in the pretreatment solution I, drain the remaining pretreatment solution I on absorbent paper, and then immerse the tissue sections in the pretreatment solution II preheated to 72°C for 10 min, The slide was shaken up and down every 2-3 minutes; the pretreatment solution II w...

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Abstract

The invention belongs to the biotechnical field, and concretely relates to a pretreatment method and a pretreatment liquid used for fluorescence in situ hybridization of FFPE tissue slices. The pretreatment liquid comprises a pretreatment liquid I and a pretreatment liquid II; the component of the pretreatment I is alkylphenol polyoxyethylene (TX-10) and / or octylphenoxypolyethoxyethanol; and components of the pretreatment liquid II comprise alkylphenol polyoxyethylene (TX-10) and / or octylphenoxypolyethoxyethanol, and also comprise ethylene diamine tetraacetic acid (EDTA) and / or a citric acid buffer. Paraffin is directly dissolved and emulsified in aqueous environment with the temperature being higher than the melting point of the paraffin through the pretreatment liquid I and the pretreatment liquid II by using the hydrophilic and lipophilic characteristics of an emulsifier in order to complete hydration and permeation, so steps are simplified, and time is saved. The pretreatment liquid II also can destroy formalin fixation induced cross-linking among proteins, so the FFPE tissues can be easily digested by proteases, and the autofluorescence and the background of the tissues are substantially reduced.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a pretreatment method and a pretreatment solution for fluorescence in situ hybridization FFPE tissue sections. Background technique [0002] Fluorescence in situ hybridization is a non-radioactive molecular genetic technique developed on the basis of radioactive in situ hybridization in the 1980s. It is a new in situ hybridization method formed by replacing radioactive isotope labels with fluorescein labels. FISH has the advantages of safety, rapidity, high sensitivity and simultaneous display of multiple colors. [0003] Using traditional methods in fluorescence in situ hybridization experiments usually requires steps such as dewaxing, rehydration, sodium thiocyanate treatment, protease digestion, 2×SSC rinsing, and alcohol dehydration and drying of FFPE sample sections. The whole process is complicated and time-consuming. longer ( figure 1 ), in addition, traditional ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6841
Inventor 晏星李雪梅叶伦程弘夏陈刚
Owner WUHAN HEALTHCHART BIOLOGICAL TECH
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