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With much of what we know about the pathogenesis of disease coming from the systematic and careful study of histological material, histology has proven to be an incredibly valuable area of study. In "Histology Protocols", expert researchers in the field contribute detailed methods involving the preparation of tissue, providing an overview of fixation, embedding, and processing, along with the required techniques for the retrieval of RNA from histological sections, followed by sections on routine and specialist histological staining techniques, including immuno-, lectin, and hybridization histochemistry, histological staining, as well as specific methods for the in situ identification of hypoxia and apoptosis, and details on the advances in imaging and image analysis. Written in the highly successful Methods in Molecular BiologyTM series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, "Histology Protocols" will provide molecular biologists with the basic histochemical techniques and histologists with the molecular techniques necessary to realize the potential of their resource.
Ian Darby updates the highly successful premier edition with a full panoply of new and greatly augmented state-of-the-art techniques. This valuable new edition contains basic and advanced in situ hybridization techniques-many not covered in the first edition-and includes protocols for in situ hybridization of whole-mount embryo specimens, in situ hybridization at the electron microscope level, in situ detection of DNA fragmentation in apoptosis, localization of genes to particular chromosomes, and the use of DNA or RNA probes to detect expression in cells or tissue sections. Prepared by leading investigators who have daily worked with the techniques, fine-tuning them for reliability, In Situ Hybridization Protocols, 2nd Edition, offers a completely updated and extended collection of proven and readily reproducible methods suitable for both the novice and experienced investigator. Its state-of-the-art protocols constitute the new standard methods resource in the field, one that will enable researchers successfully to enhance and improve their current experimental repertoire.
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 BiologyTM 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.
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