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Recent advances in fish cytogenetics have enhanced the interest in chromosome analysis in both fundamental (systematics and comparative genomics among fishes and other vertebrate groups) and applied (aquaculture, conservation and response to pollutants, whole genome sequencing of model fish species) research. Although the genomic material, the chro
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.
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.
Cytogenetics is the study of chromosome morphology, structure, pathology, function, and behavior. The field has evolved to embrace molecular cytogenetic changes, now termed cytogenomics. Cytogeneticists utilize an assortment of procedures to investigate the full complement of chromosomes and/or a targeted region within a specific chromosome in metaphase or interphase. Tools include routine analysis of G-banded chromosomes, specialized stains that address specific chromosomal structures, and molecular probes, such as fluorescence in situ hybridization (FISH) and chromosome microarray analysis, which employ a variety of methods to highlight a region as small as a single, specific genetic sequence under investigation. The AGT Cytogenetics Laboratory Manual, Fourth Edition offers a comprehensive description of the diagnostic tests offered by the clinical laboratory and explains the science behind them. One of the most valuable assets is its rich compilation of laboratory-tested protocols currently being used in leading laboratories, along with practical advice for nearly every area of interest to cytogeneticists. In addition to covering essential topics that have been the backbone of cytogenetics for over 60 years, such as the basic components of a cell, use of a microscope, human tissue processing for cytogenetic analysis (prenatal, constitutional, and neoplastic), laboratory safety, and the mechanisms behind chromosome rearrangement and aneuploidy, this edition introduces new and expanded chapters by experts in the field. Some of these new topics include a unique collection of chromosome heteromorphisms; clinical examples of genomic imprinting; an example-driven overview of chromosomal microarray; mathematics specifically geared for the cytogeneticist; usage of ISCN’s cytogenetic language to describe chromosome changes; tips for laboratory management; examples of laboratory information systems; a collection of internet and library resources; and a special chapter on animal chromosomes for the research and zoo cytogeneticist. The range of topics is thus broad yet comprehensive, offering the student a resource that teaches the procedures performed in the cytogenetics laboratory environment, and the laboratory professional with a peer-reviewed reference that explores the basis of each of these procedures. This makes it a useful resource for researchers, clinicians, and lab professionals, as well as students in a university or medical school setting.
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.
In the past 20 years, fish cytogenetics has become an essential tool in fields as diverse as systematics and evolution, conservation, aquaculture and more recently, genomics. This book is organized in four sections (systematics and evolution; biodiversity conservation; stock assessment and aquaculture; genomics) covering the major fields of present fish cytogenetic research. The eighteen contributions from thirteen countries which make up this book, provide a comprehensive picture of the ongoing research around the world. Due to the diversified arrays of themes approached, including speciation and evolution, biodiversity and conservation and genomics, the book is addressed not only to specialists in cytogenetics but to all scientists interested in fish biology.
This reference book provides information on plant cytogenetics for students, instructors, and researchers. Topics covered by international experts include classical cytogenetics of plant genomes; plant chromosome structure; functional, molecular cytology; and genome dynamics. In addition, chapters are included on several methods in plant cytogenetics, informatics, and even laboratory exercises for aspiring or practiced instructors. The book provides a unique combination of historical and modern subject matter, revealing the central role of plant cytogenetics in plant genetics and genomics as currently practiced. This breadth of coverage, together with the inclusion of methods and instruction, is intended to convey a deep and useful appreciation for plant cytogenetics. We hope it will inform and inspire students, researchers, and teachers to continue to employ plant cytogenetics to address fundamental questions about the cytology of plant chromosomes and genomes for years to come. Hank W. Bass is a Professor in the Department of Biological Science at Florida State University. James A. Birchler is a Professor in the Division of Biological Sciences at the University of Missouri.
Following a section on tissue culture, chromosome staining and basic information about karyotyping, this text presents nomenclature and quality standards, as well as protocols of relevance to comprehensive cytogenetic diagnostics.