Journal of Molecular Histology

, Volume 38, Issue 2, pp 151–157 | Cite as

Fluorescent in situ hybridization on tissue microarrays: challenges and solutions

Original Paper

Abstract

Tissue microarray (TMA) technology has provided a high throughput means of evaluating potential biomarkers and therapeutic targets in archival pathological specimens. TMAs facilitate the rapid assessment of molecular alterations in hundreds of different tumours on a single slide. Sections from TMAs can be used for any in situ tissue analysis, including fluorescent in situ hybridization (FISH). FISH is a molecular technique that detects numerical and structural abnormalities in both metaphase chromosomes and interphase nuclei. FISH is commonly used as a prognostic and diagnostic tool for the detection of translocations and for the assessment of gene deletion and amplification in tumours. Performing FISH on TMAs enables researchers to determine the clinical significance of specific genetic alterations in hundreds of highly characterized tumours. The use of FISH on archival paraffin embedded tissues is technically demanding and becomes even more challenging when applied to paraffin embedded TMAs. The problems encountered with FISH on TMAs, including probe preparation, hybridization, and potential applications of FISH, will be addressed in this review.

Keywords

Fluorescent in situ hybridization Tissue microarray Formalin fixed paraffin embedded tissue 

References

  1. Andersen CL et al (2001) Improved procedure for fluorescence in situ hybridization on tissue microarrays. Cytometry 45(2):83–86PubMedCrossRefGoogle Scholar
  2. Brown LA et al (2006) Amplification of EMSY, a novel oncogene on 11q13, in high grade ovarian surface epithelial carcinomas. Gynecol Oncol 100(2):264–270PubMedCrossRefGoogle Scholar
  3. Chin SF et al (2003) A simple and reliable pretreatment protocol facilitates fluorescent in situ hybridisation on tissue microarrays of paraffin wax embedded tumour samples. Mol Pathol 56(5):275–279PubMedCrossRefGoogle Scholar
  4. dos Santos NR et al (2001) Molecular mechanisms underlying human synovial sarcoma development. Genes Chromosomes Cancer 30(1):1–14PubMedCrossRefGoogle Scholar
  5. Ensinger C et al (1997) Improved technique for investigations on archival formalin-fixed, paraffin-embedded tumors by interphase in-situ hybridisation. Anticancer Res 17(6D):4633–4637PubMedGoogle Scholar
  6. Feinberg AP, Vogelstein B (1983) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132(1):6–13PubMedCrossRefGoogle Scholar
  7. Feinberg AP, Vogelstein B (1984) A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Addendum. Anal Biochem 137(1):266–267PubMedCrossRefGoogle Scholar
  8. Fisher C (1998) Synovial sarcoma. Ann Diagn Pathol 2(6):401–421PubMedCrossRefGoogle Scholar
  9. Fletcher JA (1999) DNA in situ hybridization as an adjunct in tumor diagnosis. Am J Clin Pathol 112(1 Suppl 1):S11–S18PubMedGoogle Scholar
  10. Hughes-Davies L et al (2003) EMSY links the BRCA2 pathway to sporadic breast and ovarian cancer. Cell 115(5):523–535PubMedCrossRefGoogle Scholar
  11. Kearney L (1999) The impact of the new fish technologies on the cytogenetics of haematological malignancies. Br J Haematol 104(4):648–658PubMedCrossRefGoogle Scholar
  12. Knuutila S et al (1998) DNA copy number amplifications in human neoplasms: review of comparative genomic hybridization studies. Am J Pathol 152(5):1107–1123PubMedGoogle Scholar
  13. Kononen J et al (1998) Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nat Med 4(7):844–847PubMedCrossRefGoogle Scholar
  14. Lee CH et al (2005) Assessment of Her-1, Her-2, And Her-3 expression and Her-2 amplification in advanced stage ovarian carcinoma. Int J Gynecol Pathol 24(2):147–152PubMedCrossRefGoogle Scholar
  15. Makretsov N et al (2004) A fluorescence in situ hybridization study of ETV6-NTRK3 fusion gene in secretory breast carcinoma. Genes Chromosomes Cancer 40(2):152–157PubMedCrossRefGoogle Scholar
  16. Pergament E et al (2000) The clinical application of interphase FISH in prenatal diagnosis. Prenat Diagn 20(3):215–220PubMedCrossRefGoogle Scholar
  17. Prentice LM et al (2005) NRG1 gene rearrangements in clinical breast cancer: identification of an adjacent novel amplicon associated with poor prognosis. Oncogene 24(49):7281–7289PubMedCrossRefGoogle Scholar
  18. Rigby PW et al (1977) Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol 113(1):237–251PubMedCrossRefGoogle Scholar
  19. Schraml P et al (1999) Tissue microarrays for gene amplification surveys in many different tumor types. Clin Cancer Res 5(8):1966–1975PubMedGoogle Scholar
  20. Spiridon CI et al (2002) Targeting multiple Her-2 epitopes with monoclonal antibodies results in improved antigrowth activity of a human breast cancer cell line in vitro and in vivo. Clin Cancer Res 8(6):1720–1730PubMedGoogle Scholar
  21. Terry J et al (2005) Fluorescence in situ hybridization for the detection of t(X;18)(p11.2;q11.2) in a synovial sarcoma tissue microarray using a breakapart-style probe. Diagn Mol Pathol 14(2):77–82PubMedCrossRefGoogle Scholar
  22. Tomlins SA et al (2005) Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 310(5748):644–648PubMedCrossRefGoogle Scholar
  23. Tomlins SA et al (2006) TMPRSS2:ETV4 gene fusions define a third molecular subtype of prostate cancer. Cancer Res 66(7):3396–3400PubMedCrossRefGoogle Scholar
  24. Yoshimoto M et al (2006) Three-color FISH analysis of TMPRSS2/ERG fusions in prostate cancer indicates that genomic microdeletion of chromosome 21 is associated with rearrangement. Neoplasia 8(6):465–469PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2007

Authors and Affiliations

  1. 1.Genetic Pathology Evaluation Centre of the Prostate CentreUniversity of British ColumbiaVancouverCanada
  2. 2.Department of Pathology of Vancouver Coastal Health Research Institute, British Columbia Cancer AgencyUniversity of British ColumbiaVancouverCanada
  3. 3.Department of Pathology, Center for Translational and Applied Genomics, British Columbia Cancer AgencyUniversity of British ColumbiaVancouverCanada

Personalised recommendations