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.




Metal-Enhanced Fluorescence


Book Description

Discover how metal-enhanced fluorescence is changing traditional concepts of fluorescence This book collects and analyzes all the current trends, opinions, and emerging hot topics in the field of metal-enhanced fluorescence (MEF). Readers learn how this emerging technology enhances the utility of current fluorescence-based approaches. For example, MEF can be used to better detect and track specific molecules that may be present in very low quantities in either clinical samples or biological systems. Author Chris Geddes, a noted pioneer in the field, not only explains the fundamentals of metal-enhanced fluorescence, but also the significance of all the most recent findings and models in the field. Metal-enhanced fluorescence refers to the use of metal colloids and nanoscale metallic particles in fluorescence systems. It offers researchers the opportunity to modify the basic properties of fluorophores in both near- and far-field fluorescence formats. Benefits of metal-enhanced fluorescence compared to traditional fluorescence include: Increased efficiency of fluorescence emission Increased detection sensitivity Protect against fluorophore photobleaching Applicability to almost any molecule, including both intrinsic and extrinsic chromophores Following a discussion of the principles and fundamentals, the author examines the process and applications of metal-enhanced fluorescence. Throughout the book, references lead to the primary literature, facilitating in-depth investigations into particular topics. Guiding readers from the basics to state-of-the-technology applications, this book is recommended for all chemists, physicists, and biomedical engineers working in the field of fluorescence.




RNA Tagging


Book Description

This book provides a compendium of state-of-the-art methods for the labeling, detection, and purification of RNA and RNA-protein complexes and thereby constitutes an important toolbox for researchers interested in understanding the complex roles of RNA molecules in development, signaling, and disease. Beginning with a section on in situ detection of RNA molecules using FISH techniques, the volume continues with parts exploring in vivo imaging of RNA transport and localization, imaging and analysis of RNA uptake and transport between cells, identification and analysis of RNA-binding proteins, guide RNAs in genome editing, as well as other specific analytical techniques. Written for the highly successful Methods in Molecular Biology series, 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, RNA Tagging: Methods and Protocols serves as a vital reference for researchers looking to further the increasingly important research in RNA biology.




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.




Handbook of Immunohistochemistry and in Situ Hybridization of Human Carcinomas


Book Description

The various cell types have traditionally been recognized and classified according to their appearance in the light microscope following the process of fixing, processing, sectioning, and staining tissues that is known as histology. Classical histology has been augmented by immunohistochemistry (the use of specific antibodies to stain particular molecular species in situ). Immunohistochemistry has allowed the identification of many more cell types than could be visualized by classical histology, particularly in the immune system and among the scattered hormone-secreting cells of the endocrine system. Handbook of Immunohistochemistry and in Situ Hybridization of Human Carcinomas discusses all aspects of immunohistochemistry and in situ hybridization technologies and the important role they play in reaching a cancer diagnosis. It provides step-by-step instructions on the methods of additional molecular technologies such as DNA microarrays, and microdissection, along with the benefits and limitations of each method. The topics of region-specific gene expression, its role in cancer development and the techniques that assist in the understanding of the molecular basis of disease are relevant and necessary in science today, ensuring a wide audience for this book. - The only book available that translates molecular genetics into cancer diagnosis - Provides the readers with tools necessary to perform and optimize sensitive, powerful techniques, including immunohistochemistry and fluorescence in situ hybridization, used in tumor diagnosis - Written by experts in this field, the book provides theoretical considerations as well as practical approaches to carry out effectively these techniques - Offers suggestions, tips, cautions, and guidelines to avoid artifacts and misdiagnosis - Introduces new techniques to detect genes and proteins involved in the initiation and progression of cancer - Covers the latest developments and a wide range of applications to the detection of antigens and single-copy DNA and RNA - Written in a uniform format, each chapter includes Introduction, Materials required, step-by-step detailed Methods, Results, Discussion, and comprehensive up-to-date References




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.




Techniques and Methods in Biology


Book Description

This comprehensive and accessible text is aimed at undergraduate and postgraduate students of biotechnology, and postgraduate students of botany and zoology. The book discusses spectroscopy, which forms the core of modern research, and microscopy, which is now an indispensable analytical tool. It also illustrates radioactivity and related phenomena, and evaluates the role of statistics.




In Situ Hybridization Protocols


Book Description

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.




Radiative Decay Engineering


Book Description

During recent years our enthusiasm for this field has continually increased. This book presents expert contributions describing the fundamental principles for the widespread use of radiative decay engineering in the biological sciences and nanotechnology.




ICRF Handbook of Genome Analysis


Book Description

The combined power of genetic analysis and recombinant DNA technology to analyse entire genomes has moved biomedical research into a new and revolutionary phase. The complete sequencing and mapping of the human genome, as well as the genomes of other model organisms, will be the basis for our future understanding of human disease, and will allow us to answer fundamental questions about development and evolution. T The new ICRF Handbook of Genome Analysis is the essential guide to the enormous range of techniques available to the researcher for both the genetic and physical mapping of the genome, as well as the sequencing and analysis of DNA. It is both a protocol manual and a comprehensive information resource. Written by international experts, each chapter presents a state-of-the-art review of a methodology. Methods are fully described and evaluated; their advantages and disadvantages discussed; and their suitability for different investigations considered. Step-by-step protocols, including computer analyses, are given for 123 essential experimental procedures. 'Troubleshooting' sections discuss possible reasons for failure and offer remedies. The primary focus is on human genetics and the benefits of an understanding of the genome for the diagnosis and treatment of human disease. The book also considers the current state of progress in the analysis of genomes of many model organisms, including plants. A major part of the work provides detail on Internet resources as well as basic data on human and other genomes, including mapped disease genes and mouse knockouts. Covers not only the human genome in relation to cancers and other human diseases, but also the genomes of all important model organisms Contains 123 easy-to-follow protocols for essential experimental procedures Reviews a vast range of other information resources, including journals and the Internet * provides an invaluable listing of suppliers of laboratory materials Has been written by international experts from their own practical experience Is mandated by the Imperial Cancer Research Fund - a leader in research in this field Has a sturdy spiral binding within a hardback case for ease of use in the lab