Fluorescent In Situ Hybridization Techniques in Pathology: Principle, Technique and Applications
Fluorescent in situ hybridization (FISH) technique is a popular molecular technique. Here the double-stranded DNA is at first converted into single-stranded DNA, and then subsequently a fluorescent-tagged probe is used to visualize the target DNA part. It is possible to do FISH on the paraffin-embedded tissue material, and therefore it can be used in archival materials. FISH technique bypasses the tedious cell culture technique. Most importantly cytogenetic abnormality of the cells can be demonstrated along with the morphology of the cell, and these two can be correlated. The chapter describes the basic principles, advantages, limitations, protocols and applications of FISH. In addition, troubleshooting of FISH has been also described here. Comparative genomic hybridization (CGH) provides the information on global view of gain or loss of chromosome of the tumor genome. The present chapter elucidates the basic principle, protocol and applications of CGH. The variant of CGH is array-based CGH (aCGH). In aCGH a specific target DNA sequence is used instead of metaphase chromosome. Microarray plate with multiple wells contains genomic bacterial artificial chromosome or cDNA in the array. The basic steps of aCGH have also been described here.
KeywordsFluorescent in situ hybridization technique FISH Troubleshooting of FISH Protocol of FISH Comparative genomic hybridization CGH Multi-coloured FISH Spectral karyotyping Three-dimensional FISH Living cell cytogenetic array-based CGH
- 13.DeVries S, Gray JW, Pinkel D, Waldman FM, Sudar D. Comparative genomic hybridization. Curr Protoc Hum Genet. 2001;Chapter 4:Unit4.6.Google Scholar
- 14.Lucito R, Healy J, Alexander J, Reiner A, Esposito D, Chi M, Rodgers L, Brady A, Sebat J, Troge J, West JA, Rostan S, Nguyen KC, Powers S, Ye KQ, Olshen A, Venkatraman E, Norton L, Wigler M. Representational oligonucleotide microarray analysis: a high-resolution method to detect genome copy number variation. Genome Res. 2003;13(10):2291–305.CrossRefPubMedPubMedCentralGoogle Scholar