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Chromosomal analysis of non-small-cell lung cancer by Fluorescence in situ hybridisation (FISH) (CAT#: STEM-MB-1186-WXH)

Introduction

Lung cancer is the biggest cause of cancer mortality in the world and has one of the worst survival rates of any solid tumour. Around 75–80% of all cases of lung cancer diagnosed are non-small-cell lung cancer (NSCLC). <br />Multicolour fluorescent in situ hybridisation (M-FISH) is a karyotyping technique that uses whole chromosome paints (WCP) to label each chromosome a unique colour. Use of this technique facilitates the karyotyping of tumour cells and allows the identification of marker chromosomes and complex chromosomal rearrangements that would previously have been uninterpretable.




Principle

FISH uses fluorescent probes with complementary base sequences to locate the presence or absence of specific portions of DNA on chromosomes. The probe and target DNA must be denatured with heat or chemicals to break hydrogen bonds in the DNA and to allow hybridisation to occur once the two samples are mixed. The fluorescent probes form new hydrogen bonds with their complementary base pairs on the DNA, and these can then be detected via microscopy.

Applications

Detect and localize the presence or absence of specific DNA sequences on chromosomes.
Detect and localize specific RNA targets (mRNA, lncRNA and miRNA) in cells, circulating tumor cells, and tissue samples.

Procedure

1. Sample preparation
2. Co-denaturation and hybridization
3. Probe detection
4. Wash off of unbound probe
5. Analysis by flow cytometer/fluorescence microscopy

Materials

• Flow cytometer
• Fluorescence microscopy