In Situ Hybridization Protocols

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.

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.