In Situ Hybridization Protocols

ISBN-13: 9781617375446 / Angielski / Miękka / 2010 / 270 str.

Ian A. Darby; Tim D. Hewitson

Opis

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.

Recenzje

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 in the 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)

Spis treści

Treatment of Tissue Sections for In Situ Hybridization Gregory H. Tesch, Hui Y. Lan, and David J. Nikolic-Paterson Preparation of Template DNA and Labeling Techniques Peter J. Roche In Situ Hybridization Using cRNA Probes: Isotopic and Nonisotopic Detection Methods Ian A. Darby, Teresa Bisucci, Alexis Desmoulière, and Tim D. Hewitson Tyramide Signal Amplification for DNA and mRNA In Situ Hybridization Ernst J. M. Speel, Anton H. N. Hopman, and Paul Komminoth Expression Analysis of Murine Genes Using In Situ Hybridization With Radioactive and Nonradioactively Labeled RNA Probes Anne Chotteau-Lelièvre, Pascal Dollé, and Françoise Gofflot Nonradioactive In Situ Hybridization on Frozen Sections and Whole Mounts Julia Asp, Alexandra Abramsson, and Christer Betsholtz. In Situ Hybridization of Whole-Mount Embryos Murray Hargrave, Josephine Bowles, and Peter Koopman In Situ Detection of Epstein-Barr Virus and Phenotype Determination of EBV-Infected Cells Gerald Niedobitek and Hermann Herbst 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, and Tsuguhiro Kaneda Electron Microscopy and In Situ Hybridization: Expression of P2Y2 Receptor mRNA in the Cerebellum Andrzej Loesch and Rainer Glass In Situ Hybridization Using Riboprobes on Free-Floating Brain Sections Neil C. Owens, F. Martin Hess, and Emilio Badoer An In Situ Hybridization Technique to Detect Low-Abundance Slug mRNA in Adherent Cultured Cells Changsun Choi, Laurie G. Hudson, Pierre Savagner, and Donna F. Kusewitt Identification of Transplanted Human Cells in Animal Tissues Daniel Benten, Kang Cheng, and Sanjeev Gupta In Situ Hybridization to Plant Tissues and Chromosomes AndreasHouben, Sharon J. Orford, and Jeremy N. Timmis Histochemical Localization of Cell Proliferation Using In Situ Hybridization for Histone mRNA Tim D. Hewitson, Kristen J. Kelynack, and Ian A. Darby Histochemical Localization of Apoptosis With In Situ Labeling of Fragmented DNA Tim D. Hewitson, Teresa Bisucci, and Ian A. Darby The Use of Combined Immunohistochemical Labeling and In Situ Hybridization to Colocalize mRNA and Protein in Tissue Sections Malcolm D. Smith, Michael Ahern, and Mark Coleman Semiquantitative In Situ Hybridization Using Radioactive Probes to Study Gene Expression in Motoneuron Populations Paul D. Storer and Tracey DeLucia Quantitative In Situ Hybridization of Tissue Microarrays Adrian M. Jubb, Thinh Q. Pham, Gretchen D. Frantz, Franklin V. Peale, Jr., and Kenneth J. Hillan Index

O autorze

In Situ Hybridization has successfully provided basic and advanced laboratory techniques. With increasing technology and evolving applications, this edition concentrates on in situ hybridization of cells and tissues and the derivative techniques that have not been covered in the previous editions. The third edition will be a valuable aid in setting up the technique or improving the sensitivity and scope of applications of in situ hybridization in their own laboratories.

Blurb

In In Situ Hybridization, Third Edition, Ian Darby updates his highly successful second edition and shifts the focus to tissue and cell in situ hybridization. Drawing on experts working in diverse areas and taking advantage of advances in non-isotopic in situ hybridization and amplification techniques, the authors describe in detail their readily reproducible techniques for preparation, detection, and tissue hybridization. Among the new techniques detailed are PNA probes for viral diagnostics, plant in situ hybridization, cell proliferation detection, and quantitation of in situ hybridization. There are also cutting-edge techniques for tissue microarrays, expanded embryology-developmental gene detection, and expanded cell culture. Derivative techniques presented include the identification of transplanted cells, histones, nick-end labeling for apoptosis, the use of peptide nucleic acid probes, and in situ hybridization of plant specimens. The protocols follow the successful Methods in Molecular Biology™ series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.
Completely updated and highly practical, In Situ Hybridization, Third Edition allows both the molecular biologist with little experience of histology and the histologist with little experience of molecular biology to use these powerful new techniques successfully.