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.

In Situ Hybridization Protocols, Fourth Edition contains 21 protocols that utilize the in situ hybridization technology to document or take advantage of the visualization of specific RNA molecules. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, In Situ Hybridization Protocols, Fourth Edition seeks to aid scientists in the further discovery of new RNA species and uncovering of their cellular functions.

Molecular biology and genetic engineering techniques have revolutionized our concepts of gene activity in cellular function. In situ hybridization has contributed to this revolution. It is the only technique which gives precise data on the content and distribution of DNA and mRNA in intactcells and tissues. It is now an invaluable test in understanding the pathophysiology of cellular functions. This book covers all aspects of in situ hybridization. It describes practical procedures and protocols, the scientific background, areas of application, and the limitations of the technology. The contributing authors are all experts. Each chapter contains up-to-date references, many colourillustrations, the latest advances, and prospects for the future. This practical guide will be invaluable to: molecular, cell, and developmental biologists; cyto- and molecular geneticists; clinicians, especially pathologists, biologists, and microbiologists; and scientists seriously interested inthe pathophysiology of cell function and its regulation.

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 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 book provides detailed information on basic and advanced laboratory techniques in histopathology and cytology. It discusses the principles of and offers clear guidance on all routine and special laboratory techniques. In addition, it covers various advanced laboratory techniques, such as immunocytochemistry, flow cytometry, liquid based cytology, polymerase chain reaction, tissue microarray, and molecular technology. Further, the book includes numerous color illustrations, tables and boxes to familiarize the reader with the work of a pathology laboratory. The book is mainly intended for postgraduate students and fellows in pathology as well as practicing pathologists. The book is also relevant for all the laboratory technicians and students of laboratory technology.

John Walker and Ralph Rapley have collected a wide-ranging group of molecular and biochemical techniques that are the most frequently used in medical and clinical research, especially diagnostics. The authors-well-established investigators who run their own research programs and use the methods on a regular basis-outline the practical procedures for using them and describe a variety of pertinent applications. Among the technologies presented are southern and western blotting, electrophoresis, PCR, cDNA and protein microarrays, liquid chromatography, in situ hybridization, karyotyping, flow cytometry, bioinformatics, genomics, and ribotyping. The applications include assays for mutation detection, mRNA analysis, chromosome translocations, inborn errors of metabolism, protein therapeutics, and gene therapy.

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 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.