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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
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.
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.
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.
Fluorescence in situ hybridization (FISH) has been developed as a powerful technology which allows direct visualisation or localisation of genomic alterations. The technique has been adopted to a range of applications in both medicine, especially in the areas of diagnostic cytogenetics, and biology. Topics described in this manual include: FISH on native human tissues, such as blood, bone marrow, epithelial cells, hair root cells, amniotic fluid cells, human sperm cells; FISH on archival human tissues, such as formalin fixed and paraffin embedded tissue sections, cryofixed tissue; simultaneous detection of apoptosis and xpression of apoptosis-related genes; comparative genomic ybridization; and special FISH techniques.
This book is a unique source of information on the present state of the exciting field of molecular cytogenetics and how it can be applied in research and diagnostics. The basic techniques of fluorescence in situ hybridization and primed in situ hybridization (PRINS) are outlined, the multiple approaches and probe sets that are now available for these techniques are described, and applications of them are presented in 36 chapters by authors from ten different countries around the world. The book not only provides the reader with basic and background knowledge on the topic, but also gives detailed protocols that show how molecular cytogenetics is currently performed by specialists in this field. The FISH Application Guide initially provides an overview of the (historical) development of molecular cytogenetics, its basic procedures, the equipment required, and probe generation. The book then describes tips and tricks for making different tissues available for molecular cytogenetic studies. These are followed by chapters on various multicolor FISH probe sets, their availability, and their pot- tial for use in combination with other approaches. The possible applications that are shown encompass the characterization of marker chromosomes, cryptic cytogenetic aberrations and epigenetic changes in humans by interphase and metaphase cyto- netics, studies of nuclear architecture, as well as the application of molecular cytogenetics to zoology, botany and microbiology.
In the era of precision medicine, physicians are increasingly in need of more definitive diagnostic, prognostic, and predictive information derived from small biopsy specimens such as cytology samples in order to guide effective patient care. Cytopathology is well poised to meet this challenge. Whilst the traditional cytomorphologic component of cytology practice is still valid, enormous advances have been made in the field of cytopathology thanks to transformative technology and innovative individuals that have augmented the cytologists' ability to meet the demands of modern medicine. The purpose of this book is to describe, illustrate, and review many of the most recent developments regarding modern techniques employed in cytopathology. This latest monograph is intended for all cytologists including cytopathologists, cytotechnologists, cytology lab assistants, trainees, research scientists, and anyone who is interested in the field of cytopathology. We have invited pioneering experts in their respective fields to author these chapters. This book is not only the culmination of their groundbreaking work and effort but also presents a critical review of the current literature. We have attempted to provide readers with an informative and comprehensive aid so that they may better appreciate how emerging technology has been applied to cytology. Each chapter in this book presents a stand-alone contemporary review of emerging topics in cytopathology. We hope that you will find this monograph thought-provoking and a valuable reference for your practice.
This volume contains a comprehensive compilation of chromogenic and fluorescent RNA in situ hybridization (ISH) technology in many of its various shades, forms, and applications. The book is organized into a number of parts and chapters focusing on the application of ISH methodologies to different animal species as used in Evolutionary Development (EvoDevo) and Biomedical research, and covering new developments in RNA visualization by fluorescent ISH (FISH). The described (F)ISH protocols employ effective strategies for signal enhancement and target amplification allowing for high signal intensities and drastically improved signal-to-noise ratios. Chromogenic and fluorescent ISH, as specified in the various chapters, are most essential for RNA expression profiling, applied to many fields of research including cellular, developmental, and evolutionary biology, neurobiology and neuropathology. Written for the popular Neuromethods series, chapters include the kind of detail and key implementation advice that ensures successful results in the laboratory. Essential and authoritative, In Situ Hybridization Methods provides detailed protocols for newcomers to ISH, and inspires researchers familiar with the technique to seek and find up-to-date methodology for new and specialized applications.