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 field of biotechnology, and in particular relates to a pretreatment method and a pretreatment solution for fluorescence in situ hybridization FFPE tissue sections. Described pretreatment liquid comprises pretreatment liquid I and pretreatment liquid II; The component of described pretreatment liquid I is alkylphenol polyoxyethylene ether (TX-10) and / or ethylphenyl polyethylene glycol; The components of the pretreatment liquid II include alkylphenol polyoxyethylene ether (TX‑10) and / or ethylphenyl polyethylene glycol, and also include ethylenediaminetetraacetic acid (EDTA) and / or citric acid buffer liquid. The present invention uses the pretreatment liquid I and the pretreatment liquid II to directly dissolve the emulsified paraffin in an aqueous environment higher than the melting point of the paraffin by using the hydrophilic and lipophilic characteristics of the emulsifier, completing the hydration and penetration, simplifying the steps and saving time. Pretreatment solution II can also break the interprotein cross-links caused by formaldehyde fixation, making FFPE tissues easier to be digested by proteases, and significantly reducing tissue autofluorescence and background.

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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