In Situ Hybridization Protocols

by Darby, Ian A.; Hewitson, Tim D.

• ISBN13: 9781588294029
• ISBN10: 1588294021
• Edition: 3rd
• Format: Hardcover
• Copyright: 2006-01-30
• Publisher: Humana Pr Inc

Summary

This revised and updated edition of a recognized classic emphasizes tissue and cell in situ hybridization methods. 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 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.

Table of Contents

Preface	v
Contributors	ix
1 Treatment of Tissue Sections for In Situ Hybridization
Gregory H. Tesch, Hui Y. Lan, and David J. Nikolic-Paterson	1	(8)
2 Preparation of Template DNA and Labeling Techniques
Peter J. Roche	9	(8)
3 In Situ Hybridization Using cRNA Probes: Isotopic and Nonisotopic Detection Methods
Ian A. Darby, Teresa Bisucci, Alexis Desmoulière, and Tim D. Hewitson	17	(16)
4 Tyramide Signal Amplification for DNA and mRNA In Situ Hybridization
Ernst J.M. Speel, Anton H.N. Hopman, and Paul Komminoth	33	(28)
5 Expression Analysis of Murine Genes Using In Situ Hybridization With Radioactive and Nonradioactively Labeled RNA Probes
Anne Chotteau-Lelièvre, Pascal Dollé, and Françoise Gofflot	61
6 Nonradioactive In Situ Hybridization on Frozen Sections and Whole Mounts
Julia Asp, Alexandra Abramsson, and Christer Betsholtz	89	(14)
7 In Situ Hybridization of Whole-Mount Embryos
Murray Hargrave, Josephine Bowles, and Peter Koopman	103	(12)
8 In Situ Detection of Epstein—Barr Virus and Phenotype Determination of EBV-Infected Cells
Gerald Niedobitek and Hermann Herbst	115	(24)
9 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	139	(12)
10 Electron Microscopy and In Situ Hybridization: Expression of P2Y2 Receptor mRNA in the Cerebellum
Andrzej Loesch and Rainer Glass	151	(12)
11 In Situ Hybridization Using Riboprobes on Free-Floating Brain Sections
Neil C. Owens, F. Martin Hess, and Emilio Badoer	163	(10)
12 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	173	(16)
13 Identification of Transplanted Human Cells in Animal Tissues
Daniel Benten, Kang Cheng, and Sanjeev Gupta	189	(14)
14 In Situ Hybridization to Plant Tissues and Chromosomes
Andreas Houben, Sharon J. Orford, and Jeremy N. Timmis	203	(16)
15 Histochemical Localization of Cell Proliferation Using In Situ Hybridization for Histone mRNA
Tim D. Hewitson, Kristen J. Kelynack, and Ian A. Darby	219	(8)
16 Histochemical Localization of Apoptosis With In Situ Labeling of Fragmented DNA
Tim D. Hewitson, Teresa Bisucci, and Ian A. Darby	227	(8)
17 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	235	(12)
18 Semi quantitative In Situ Hybridization Using Radioactive Probes to Study Gene Expression in Motoneuron Populations
Paul D. Storer and Tracey DeLucia	247	(8)
19 Quantitative In Situ Hybridization of Tissue Microarrays
Adrian M. Jubb, Thinh Q. Pham, Gretchen D. Frantz, Franklin V. Peale, Jr., and Kenneth J. Hillan	255	(10)
Index	265