Specimen pretreatment method for reducing auto-fluorescence interference in FISH (fluorescence in situ hybridization) and reagent kit therefor

A fluorescence in situ hybridization and autofluorescence technology, used in fluorescence/phosphorescence, microbial determination/inspection, biochemical equipment and methods, etc. Background fluorescence ratio, affecting signal counting and interpretation, etc., can improve the quality and accuracy of inspection, reduce autofluorescence in human tissue, and improve signal resolution.

Active Publication Date: 2013-11-06
CHUNG GUNG MEDICAL FOUNDATION LINKOU BRANCH
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, commonly used fluorescent substances for labeling DNA or antibodies are FITC fluorescein isothiocyanate, Oregon green (Oregon green) 488, and Spectrum Green (SpectrumGreen). ) Such as: collagen, elastin, reduced nicotinamide adenine dinucleotide NADH, flavin mononucleotide (FMN, Flavin mononucleotide), flavin adenine dinucleotide (FAD, Flavin adenine dinucleotide) and aromatic Amino acids (aromatic amino acids), if the specimen has strong background autofluorescence, it wil

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Specimen pretreatment method for reducing auto-fluorescence interference in FISH (fluorescence in situ hybridization) and reagent kit therefor
  • Specimen pretreatment method for reducing auto-fluorescence interference in FISH (fluorescence in situ hybridization) and reagent kit therefor
  • Specimen pretreatment method for reducing auto-fluorescence interference in FISH (fluorescence in situ hybridization) and reagent kit therefor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0025] Embodiments are carried out on the premise of the technical solutions of the present invention, and detailed implementation methods and specific operation processes are provided, but the protection scope of the present invention is not limited to the following embodiments.

[0026] The methods used in the following examples are conventional methods unless otherwise specified.

[0027] refer to figure 1 As shown, the present invention provides a sample pretreatment method for reducing autofluorescence interference in fluorescence in situ hybridization, comprising the following steps:

[0028] Step 1 (S1): formalin-fixed paraffin-embedded (FFPE) tissue;

[0029] Step 2 (S2): slicing and drying;

[0030] Step three (S3): sodium thiocyanate (NaSCN) effect;

[0031] Step 4 (S4): Add 20 μl of 5-10 mg / mL type IV collagenase or 1 mg / mL elastase to the tissue section with an area of ​​24×32 mm that has been treated as above, and seal the slice at 37°C for 0.5-1 hr to degrade...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a specimen pretreatment method for reducing auto-fluorescence interference in FISH (fluorescence in situ hybridization) and a reagent kit therefor. The specimen pretreatment method comprises a first step of formalin fixation of the paraffin-embedded tissue, a second step of sectioning and drying, a third step of functioning of sodium thiocyanate, a fourth step of addition of 20[mu]l of collagenase type IV with the concentration of 5-10 mg/mL or elastinase with the concentration of 1mg/mL in the tissue with the area of 24*32mm and post-sealing functioning at the temperature of 37 DEG C for 0.5-1hr for degradation of the extracellular matrix with the auto-fluorescence characteristics, and a fifth step of functioning of pepsin at the temperature of 37 DEG C. The invention also provides a reagent kit for the specimen pretreatment method for reducing auto-fluorescence interference in FISH (fluorescence in situ hybridization). Through the specimen pretreatment based on the invention, the auto-fluorescence phenomena of the tissue is remarkably reduced, the ratio of fluorescent probe signals to background fluorescence is increased, and the detection quality and accuracy of FISH are improved.

Description

technical field [0001] The present invention provides a sample pretreatment method for reducing autofluorescence interference in fluorescence in situ hybridization (FISH, Fluorescence In Situ Hybridization). The treatment step adds collagenase type IV or elastase treatment step to reduce the autofluorescence phenomenon of human tissue and improve the resolution of fluorescent probe signal by fluorescence in situ hybridization. Background technique [0002] Fluorescence in situ hybridization (FISH) is a technique commonly used in clinical diagnosis, which can be used for early diagnosis, selection of treatment methods or monitoring of disease activity. For example, the treatment of breast cancer patients with trastuzumab is based on the detection of human epidermal growth factor receptor-2 (HER2) gene expression by fluorescence in situ hybridization (FISH) or when immunohistochemical staining (IHC) shows human epidermal growth factor receptor 2 (HER2) gene expression. Fluore...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12Q1/68G01N21/64
Inventor 陈泽卿黄琼瑰李尚恩徐圣琦
Owner CHUNG GUNG MEDICAL FOUNDATION LINKOU BRANCH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap