In Situ Hybridization Methods


Book Description

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.




Molecular Cytogenetics


Book Description

The new techniques of molecular cytogenetics, mainly fluorescence in situ hybridization (FISH) of DNA probes to metaphase chromosomes or interphase nuclei, have been developed in the past two decades. Many FISH techniques have been implemented for diagnostic services, whereas some others are mainly used for investigational purposes. Several hundreds of FISH probes and hybridization kits are now commercially available, and the list is growing rapidly. FISH has been widely used as a powerful diagnostic tool in many areas of medicine including pediatrics, medical genetics, maternal–fetal medicine, reproductive medicine, pathology, hematology, and oncology. Frequently, a physician may be puzzled by the variety of FISH techniques and wonder what test to order. It is not uncommon that a sample is referred to a laboratory for FISH without indicating a specific test. On the other hand, a cytogeneticist or a technologist in a laboratory needs, from case to case, to determine which procedure to perform and which probe to use for an informative result. To obtain the best results, one must use the right DNA probes and have reliable protocols and measures of quality assurance in place. Also, one must have sufficient knowledge in both traditional and molecular cytogenetics, as well as the particular areas of medicine for which the test is used in order to appropriately interpret the FISH results, and to correlate them with clinical diagnosis, treatment, and prognosis.




Molecular Pathology of Lung Diseases


Book Description

This major work, complete with 150 illustrations, many of them in color, bridges the gap between clinical pulmonary pathology and basic molecular science. Through a highly visual approach that features an abundance of tables and diagrams, the book offers a practical disease-based overview. The first two sections of the volume provide the reader with general concepts, terminology and procedures in molecular pathology. The remainder of the volume is subdivided into neoplastic and non-neoplastic lung diseases with detailed chapters covering the current molecular pathology of specific diseases. The book will be essential reading for pathologists, pulmonologists, thoracic surgeons and other health care providers interested in lung disease.




Fluorescence In Situ Hybridization (FISH) - Application Guide


Book Description

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.




FISH Technology


Book Description

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.




Chromosome Structure and Aberrations


Book Description

This book is a compilation of various chapters contributed by a group of leading researchers from different countries and covering up to date information based on published reports and personal experience of authors in the field of cytogenetics. Beginning with the introduction of chromosome, the subsequent chapters on organization of genetic material, karyotype evolution, structural and numerical variations in chromosomes, B-chromosomes and chromosomal aberrations provide an in-depth knowledge and easy understanding of the subject matter. A special feature of the book is the inclusion of a series of chapters on various types of chromosomal aberrations and their impact on breeding behaviour and crop improvement. The possible mechanism, their consequences and role in genetic analysis has been emphasized in these chapters. A few chapters have also been dedicated on various techniques routinely used in the laboratory by students and researchers. Each chapter ends with an extensive bibliography so that the students and researchers may find it relevant to consult more literature on the subject than a book of this size can offer. The book is intended to fulfill the needs of undergraduate and post graduate students of botany, zoology and agriculture besides, teachers and researchers engaged in the field of genetics, cytogenetics, and molecular genetics. In general the readers will find each chapter of the book informative and easy to understand.




Immunocytochemistry and In Situ Hybridization in the Biomedical Sciences


Book Description

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.




Basic and Advanced Laboratory Techniques in Histopathology and Cytology


Book Description

The second edition of this well-received book provides detailed information on the basic and advanced laboratory techniques in histopathology and cytology. It offers clear guidance on the principles and techniques of routine and special laboratory techniques. It also covers advanced laboratory techniques such as immunocytochemistry, flow cytometry, liquid-based cytology, polymerase chain reactions, tissue microarray, molecular technology, etc. The book's second edition covers several important recent topics with many new chapters, such as liquid biopsy, artificial neural network, digital pathology, and next-generation sequencing. Each chapter elucidates basic principle, practical methods, troubleshooting, and clinical applications of the technique. It includes multiple colored line drawings, microphotographs, and tables to illustrate each technique. The book is a helpful guide to the post-graduate students and fellows in pathology, practicing pathologists, as well as laboratory technicians, and research students.




Modern Techniques in Cytopathology


Book Description

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.




Medical BioMethods Handbook


Book Description

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.