In Situ Hybridization In Electron Microscopy

In situ hybridization is a technique that allows for the visualization of specific DNA and RNA sequences in individual cells, and is an especially important method for studying nucleic acids in heterogeneous cell populations. in situ Hybridization in Electron Microscopy reviews the three main methods developed for the ultrastructural visualization

Hybridization Techniques for Electron Microscopy examines the use of in situ hybridization techniques, including an overview of current perspectives and future developments. The book features in situ methods for fluorescence probes and confocal scanning microscopes. Three in situ hybridization methods for electron microscopes are analyzed: the non-embedded tissue method using ultrathin frozen sections, pre-embedded method, and post-embedded method using material embedded in hydrophilic resin. Positive and negative features are discussed, and clear instructions regarding implementation of techniques are provided. Particular aspects of the techniques are examined in detail, such as preparation of tissue, pretreatment, hybridization procedures, revelation (autoradiography and immunocytology) and checking procedures, in addition to the illustration, interpretation, and discussion of methods and results. The main applications described include virus detection, chromosomal gene mapping, detection of ribosomic nucleic acid, and detection of messenger RNA in animals and plants. Hybridization Techniques for Electron Microscopy is an excellent reference for cytologists, cell biologists, histochemists, cytochemists, molecular endocrinologists, and neuroendocrinologists.

In Situ hybridization allows the visualization of specific DNA/RNA sequences in individual cells in tissue sections, single cells, or chromosome preparations, and is an especially important method for studying DNA and RNA in heterogeneous cell populations. This book delves into in situ hybridization methods through the use of light microscopy used

InIn Situ Hybridization Protocols, 2nd Edition, Ian Darby updates the highly successful 1st 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. Each protocol provides detailed, easy-to-follow instructions, along with realistic commentaries on how to vary their application and how to troubleshoot problems that arise. 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.

In situ hybridization is used to reveal the location of specific nucleic acids sequences on chromosomes or in tissues. Visualization of the location of genes on chromosomes or of specific mRNAs or viruses in tissues is crucial for understanding the organization, regulation, and function of genes. It is a therefore a core technique in all areas of biomedical research. In Situ Hybridization: A Practical Approach 2/e is the second edition of one of the most successful Practical Approach books, published in 1992. Since the first edition was published, a number of important technical advances have been made. The new edition has been thoroughly updated to contain protocols detailing the major techniques of in situ hybridization currently in use: in situ hybridization to mRNA with oligonucleotide and RNA probes (radiolabelled and hapten labelled); analysis using light and electron microscopes; whole mount in situ hybridization; double detection of RNAs, and RNA plus protein; and fluorescent in situ hybridization to detect chromosomal sequences. The protocols are complemented by advice on strategies for successful results, descriptions of the theoretical basis of in situ hybridization and important new developments in gene expression databases. The procedures described are widely applicable to many systems. The use of in situ hybridization in PCR is covered in a separate volume: Herrington and O'Leary (Eds) PCR 3 - PCR in situ hybridization: A Practical Approach (OUP, 1997). All the authors have extensive practical experience of establishing reliable techniques of in situ hybridization. This book will be useful to all researchers at all levels who use in situ hybridization.

Immunocytochemistry and in situ hybridization are widely used biomedical sciences. They are essential in medical diagnosis and in cell biology research. Affinity labeling is the central goal of the experimental strategy involving a series of techniques in a logical order; from the effects of specimen fixation, through specimen preparation to expose the antigen, to optimizing immunolabeling, to assessing the result and finally to safety considerations. Numerous examples of these techniques in biomedical sciences are included, as well as experimental assays and practical tips. This survey of methods will serve as an invaluable reference source in any laboratory setting (academic, industrial or clinical) involved in research in almost every branch of biology or medicine, as well as in pharmaceutical, biotechnological and clinical applications.

Detailed protocols for analyzing DNA strand breaks, specific RNA/DNA expressions, and DNA binding proteins such as transcription factors at the individual cell level. It describes in situ nick translation and TUNEL in detail, along with radioactive and non-radioactive methods for in situ hybridization. Of special significance is the description of Southwestern histochemistry, which makes it possible to analyze the expression of transcription regulatory factors in individual cells. This volume provides an overview of the current status of molecular histochemistry along with practical how-to-do-it details. The methodology is set out in easy-to-follow language by leading scientists working in the field today. Both a rich source of the latest information and a practical lab manual.

This practical guide provides a comprehensive description of in situ hybridization, from background information to detailed and practical applications. In situ hybridization is a powerful link between cellular and molecular biology, and can be used to localize nucleic acid sequences.