Chapter 6. Tissue In Situ Hybridization

  1. Mary Hannon-Fletcher Lecturer3 and
  2. Perry Maxwell Principal Clinical Scientist4
  1. Anthony O' Grady Chief Medical Scientist1,
  2. John O' Loughlin Chief Medical Scientist2 and
  3. Hilary Magee Senior Medical Scientist2

Published Online: 11 MAY 2009

DOI: 10.1002/9780470745069.ch6

Advanced Techniques in Diagnostic Cellular Pathology

Advanced Techniques in Diagnostic Cellular Pathology

How to Cite

O' Grady, A., O' Loughlin, J. and Magee, H. (2009) Tissue In Situ Hybridization, in Advanced Techniques in Diagnostic Cellular Pathology (eds M. Hannon-Fletcher and P. Maxwell), John Wiley & Sons, Ltd, Chichester, UK. doi: 10.1002/9780470745069.ch6

Editor Information

  1. 3

    The University of Ulster, Coleraine, UK

  2. 4

    Belfast Health & Social Care Trust, Centre for Cancer Research & Cell Biology, Belfast, UK

Author Information

  1. 1

    Department of Pathology, Royal College of Surgeons in Ireland Education & Research Centre, Beaumont Hospital, Dublin, Ireland

  2. 2

    Department of Cellular Pathology, Adelaide & Meath Hospital, Dublin incorporating the National Children's Hospital, Tallaght, Dublin, Ireland

Publication History

  1. Published Online: 11 MAY 2009
  2. Published Print: 17 APR 2009

ISBN Information

Print ISBN: 9780470515976

Online ISBN: 9780470745069



  • tissue in situ hybridization;
  • Gall and Pardue and John et al and in situ hybridization (ISH) technique;
  • fluorescent in situ hybridization (FISH) - paving the way for development of simultaneous detection of multiple targets;
  • single-stranded DNA probes - generated using polymerase chain reaction (PCR);
  • biotinylated probes - first non-isotopic probes for tissue ISH;
  • ISH automation;
  • tissue preparation and ISH procedure;
  • signal amplification - tyramide signal amplification (TSA), catalysed signal amplification (CSA) and catalysed reporter deposition signal amplification (CARD);
  • clinical applications of tissue ISH and dual-colour chromogenic ISH;
  • FICTION (Fluorescence Immunophenotyping and Interphase Cytogenetics as tool for Investigation of Neoplasms) technique


The discovery of the structure of DNA in 1953 was the “sine qua non” for the development of the in situ hybridization (ISH) technique. Since then tissue ISH has developed to become a key research and diagnostic tool in the pathology laboratory. Today, a wide range of probes (both commercial and in-house) and detection systems are used to identify nucleic acid targets. Tissue ISH has become the technique of choice in determining the diagnosis and directing the treatment of many solid tumours. For example, the use of tissue ISH in the study of haematological malignancies (translocations, deletions etc.) has to a great extent superseded traditional labour-intensive karyotyping techniques. Its use in establishing the HER-2 gene status of women with breast cancer means that this population of patients can be screened rapidly and placed on the appropriate treatment regimen. The development of automation for slide pre-treatment and staining and access to a wider range of commercially available probes together with image capture and analysis software have provided the necessary standardization and reproducibility associated with other diagnostic assays. New techniques such as dual colour CISH and FICTION, the results of which can be viewed using bright-field light microscopy, have found favour amongst the pathology community because gene/protein expression can be related to tissue morphology. The continuous discovery of novel genetic signatures in a wide variety of solid tumours and the potential of these being candidates for targeted therapy combined with the aforementioned automated staining and analysis systems means that the in situ hybridization technique has a bright future in the ongoing molecular investigation of tissue samples.