In Situ Hybridization Protocols

1. Front Matter

   Pages i-xi

   PDF

2. Treatment of Tissue Sections for In Situ Hybridization

   • Gregory H. Tesch, Hui Y. Lan, David J. Nikolic-Paterson

   Pages 1-7

3. Preparation of Template DNA and Labeling Techniques

   • Peter J. Roche

   Pages 9-16

4. In Situ Hybridization Using cRNA Probes

   • Ian A. Darby, Teresa Bisucci, Alexis Desmoulière, Tim D. Hewitson

   Pages 17-31

5. Tyramide Signal Amplification for DNA and mRNA In Situ Hybridization

   • Ernst J. M. Speel, Anton H. N. Hopman, Paul Komminoth

   Pages 33-60

6. Expression Analysis of Murine Genes Using In Situ Hybridization With Radioactive and Nonradioactively Labeled RNA Probes

   • Anne Chotteau-Lelièvre, Pascal Dollé, Françoise Gofflot

   Pages 61-87

7. Nonradioactive In Situ Hybridization on Frozen Sections and Whole Mounts

   • Julia Asp, Alexandra Abramsson, Christer Betsholtz

   Pages 89-102

8. In Situ Hybridization of Whole-Mount Embryos

   • Murray Hargrave, Josephine Bowles, Peter Koopman

   Pages 103-113

9. In Situ Detection of Epstein—Barr Virus and Phenotype Determination of EBV-Infected Cells

   • Gerald Niedobitek, Hermann Herbst

   Pages 115-137

10. PNA-In Situ Hybridization Method for Detection of HIV-1 DNA in Virus-Infected Cells and Subsequent Detection of Cellular and Viral Proteins

    • Tomoko Hagiwara, Junko Hattori, Tsuguhiro Kaneda

    Pages 139-149

11. Electron Microscopy and In Situ Hybridization

    • Andrzej Loesch, Rainer Glass

    Pages 151-162

12. In Situ Hybridization Using Riboprobes on Free-Floating Brain Sections

    • Neil C. Owens, F. Martin Hess, Emilio Badoer

    Pages 163-171

13. An In Situ Hybridization Technique to Detect Low-Abundance Slug mRNA in Adherent Cultured Cells

    • Changsun Choi, Laurie G. Hudson, Pierre Savagner, Donna F. Kusewitt

    Pages 173-188

14. Identification of Transplanted Human Cells in Animal Tissues

    • Daniel Benten, Kang Cheng, Sanjeev Gupta

    Pages 189-201

15. In Situ Hybridization to Plant Tissues and Chromosomes

    • Andreas Houben, Sharon J. Orford, Jeremy N. Timmis

    Pages 203-218

16. Histochemical Localization of Cell Proliferation Using In Situ Hybridization for Histone mRNA

    • Tim D. Hewitson, Kristen J. Kelynack, Ian A. Darby

    Pages 219-226

17. Histochemical Localization of Apoptosis With In Situ Labeling of Fragmented DNA

    • Tim D. Hewitson, Teresa Bisucci, Ian A. Darby

    Pages 227-234

18. The Use of Combined Immunohistochemical Labeling and In Situ Hybridization to Colocalize mRNA and Protein in Tissue Sections

    • Malcolm D. Smith, Michael Ahern, Mark Coleman

    Pages 235-245

19. Semiquantitative In Situ Hybridization Using Radioactive Probes to Study Gene Expression in Motoneuron Populations

    • Paul D. Storer, Tracey DeLucia

    Pages 247-254

20. Quantitative In Situ Hybridization of Tissue Microarrays

    • Adrian M. Jubb, Thinh Q. Pham, Gretchen D. Frantz, Franklin V. Peale Jr., Kenneth J. Hillan

    Pages 255-264

The technique of in situ hybridization, in its various forms, has been used routinely in many laboratories for a number of years. In the post-genome era, gene arrays and proteomics have allowed us to identify hitherto unknown unrecognized pathways and mechanisms. However, rather than diminish the importance of in situ hybridization, the now widespread use of screening te- nologies has increased the need to temporally and spatially localize the dist- bution of mRNA expression. Our intention, in In Situ Hybridization Protocols is to provide ample inf- mation for novices planning to set up the in situ hybridization technique and use it in their laboratory for the first time, as well as giving updates of recent developments for those laboratories where in situ hybridization techniques are already in use. Despite its widespread significance, in situ hybridization has retained a re- tation as one of the more difficult and capricious molecular biological te- niques. This may in part be because of the hybrid nature of the technique, which often requires a mixture of molecular biological and histological skills. The two techniques are usually taught and acquired in different streams of biolo- cal science. The step-by-step and detailed protocols provided in In Situ Hybridization Protocols by researchers active in the field should make it p- sible for both the molecular biologist with little experience of histology and the histologist with little experience of molecular biology to use the technique s- cessfully in their laboratories.

• DNA
• Microarray
• Termination
• electron microscopy
• gene expression
• genes
• hybridization
• molecular biology

Reviews of Previous Editions:

"...an indispensable laboratory manual for the current techniques used...essential for the current techniques used...essential for any laboratory that is using, or wants to use, the...technique in their repertoire."-Molecular Biotechnology

" . . .a good book for anyone wishing to exploit all that in situ hybridization has to offer."
-- Biopharm Manufacturing
Reviews for Second Edition...
"This book should be of interest to anyone using or considering in situ hybridization, as it provides well written, if rather specialized, examples of the many applications of this extremely useful but notoriously tricky technique...The text is well referenced and includes useful details of reagent suppliers as well as links to further information available over the internet. This is a book that aims to make in situ hybridization accessible to all, and to inspire new applications for the technique. In my opinion, it does a reasonably sound job of both."-Heredity

From Reviews of the Second Edition...

"...highly recommended for those researchers who use in situ hybridization in their work or for those who want to exploit for the first time the possibilities that this technique offers biomedical research" -European Urology Today

"...should be available in every major biomedical library as it represents the approach that morphological scientists can use to determine gene expression under different physiological and disease conditions in mature and developing tissues." -Journal of Anatomy

From the reviews of the third edition:

"In situ hybridization is already 25 years known method, which allows to detect specific RNA and DNA sequences at the cellular level within tissue sections. The third edition of this book focuses on tissue and cell in situ hybridization. … The book is written with a broad readership in mind. It provides information for all researchers to set up this technique inthe laboratory, as well as to update the recent protocols, which are already in use." (H. Plchová, Biologia Plantarum, Vol. 50 (4), 2006)

"Darby and Hewitson have extensively changed the content and in doing so have made the book much more relevant and useful to the veterinary pathologist. The new third edition focuses on hybridization of cells and tissues, instead of having a major focus on chromosomes as in the previous edition. … reading the book cover-to-cover allows one to gain a collective appreciation for various alternatives in methodology as well as interesting innovative procedural details." (Dr. R. A. Westhouse, Veterinary Pathology, Vol. 43 (4), 2006)

• RMIT University, Melbourne, Australia

  Ian A. Darby

• The Royal Melbourne Hospital and University of Melbourne, Melbourne, Australia

  Tim D. Hewitson

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