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Presenting an area of research that intersects with and integrates diverse disciplines, including genomics, epigenetics, proteomics, and cellular biology, among others, Formalin-Fixed Paraffin-Embedded Tissues: Methods and Protocols collects contributions from expert researchers in order to provide practical guidelines to this complex study. Compiled in order to provide researchers with up-to-date methodological information pertaining to the utilization of genomic, transcriptomic, and proteomic data in diagnosis, prognosis, and tailored therapy, the ultimate aim of this volume is to decipher diseases at a molecular level. Divided into multiple convenient chapters, this detailed book covers various techniques to construct and utilize tissue arrays, it also provides detailed protocols in immunohistochemistry, immunofluorescence, fluorescent and chromogenic in situ hybridization, and ultimately introduces protocols for FFPET microdissection and nucleic acids extraction for their utilization in advanced techniques such as microarray CGH, DNA methylation and pyrosequencing. The volume also discusses FFPET research from an ethical standpoint and concludes with a chapter on novel tissue fixative. Written in the highly successful Methods in Molecular BiologyTM series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and accessible, Formalin-Fixed Paraffin-Embedded Tissues: Methods and Protocols serves as a practical guide for scientists of all backgrounds and aims to convey the appropriate sense of fascination associated with this vital field of research.
In the past decade there has been a major sea change in the way disease is diagnosed and investigated due to the advent of high throughput technologies, such as microarrays, lab on a chip, proteomics, genomics, lipomics, metabolomics etc. These advances have enabled the discovery of new and novel markers of disease relating to autoimmune disorders, cancers, endocrine diseases, genetic disorders, sensory damage, intestinal diseases etc. In many instances these developments have gone hand in hand with the discovery of biomarkers elucidated via traditional or conventional methods, such as histopathology or clinical biochemistry. Together with microprocessor-based data analysis, advanced statistics and bioinformatics these markers have been used to identify individuals with active disease or pathology as well as those who are refractory or have distinguishing pathologies. New analytical methods that have been used to identify markers of disease and is suggested that there may be as many as 40 different platforms. Unfortunately techniques and methods have not been readily transferable to other disease states and sometimes diagnosis still relies on single analytes rather than a cohort of markers. There is thus a demand for a comprehensive and focused evidenced-based text and scientific literature that addresses these issues. Hence the formulation of Biomarkers in Disease. The series covers a wide number of areas including for example, nutrition, cancer, endocrinology, cardiology, addictions, immunology, birth defects, genetics and so on. The chapters are written by national or international experts and specialists.
Methods in Enzymology volumes provide an indispensable tool for the researcher. Each volume is carefully written and edited by experts to contain state-of-the-art reviews and step-by-step protocols. In this volume, we have brought together a number of core protocols concentrating on Cell, Lipid and Carbohydrate, complementing the traditional content that is found in past, present and future Methods in Enzymology volumes. - Indispensable tool for the researcher - Carefully written and edited by experts to contain step-by-step protocols - In this volume we have brought together a number of core protocols concentrating on Cell, Lipid and Carbohydrate
A huge amount of fixed and paraffin-embedded tissue is stored in every hospital. This is very precious material that can be used for translational research and for diagnostics. The molecular methods employed for analysis of these tissues are similar to the usual molecular biology and proteomics methods, but reliable results can be obtained only if specific steps are followed with great care. This book provides detailed and precise guidelines for molecular analysis of archive tissues and will serve as an invaluable aid for researchers and pathologists involved in translational research and diagnostics. Clear notes and explanations are included to simplify use of the protocols for the less experienced. The authors are a group of acknowledged experts who have developed the described methods and validated them within the European project "Archive Tissues: Improving Molecular Medicine Research and Clinical Practice - IMPACTS", which has involved 21 leading institutions in 11 countries.
Ribonucleic acids are central to cellular and molecular processes and perform vital functions in both structural and functional roles. RNA molecules form the bridge between the stable genetic information contained within DNA and enzymes and proteins that carry out much of the metabolism within the cell. Many of the sites of protein synthesis, the ribosomes within the cell, are composed of these ribonucleic acids as are the tRNA molecules that deliver the amino acid building blocks to the ribosomes. Of all the RNA species, the nucleic acid intermediate, messenger RNA, is a desirable source of material to biologists, since this reflects much of, what ultimately, is translated into enzymes and proteins. In order to determine the qualitative and quantitative changes in mRNA expression, a vast number of molecular biological techniques have been developed. Key molecular methods that provide the means to initially isolate and analyze RNA molecules are the focus of this volume. In putting together this collection of protocols, we have tried to provide techniques that are most applicable and widely used. In particular, there are a number of iso- tion techniques included that have been developed, modified, or adapted to enable extraction from a variety of cell types, organisms, or subcellular organelles. Successful isolation of intact RNA is an essential starting point for any sub- quent analysis. This is why we have aimed to make this section comprehensive. The analysis of RNA is the focus of the following chapters.
This volume not only discusses various common biobanking topics, it also delves into less-discussed subjects such as what is needed to start a biobank, training of new biobanking personnel, and ethnic representation in biospecimen research. Other chapters in this book span practical topics including: disaster prevention and recovery; information technology; flora and fauna preservation including zoological fluid specimen photography; surgical and autopsy biobanking; biobanking of bodily fluids; biosafety; cutting frozen sections; immunohistochemistry; nucleic acid extraction; and biospecimen shipping. 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. Unique and comprehensive, Biobanking: Methods and Protocols is a valuable resource for novice and practicing biobankers, and for end-user researchers. This book aims to bring new insight into the field and expand on current biomedical biobanking studies.
This second edition provides updated and comprehensive methods on miRNA biogenesis and their role in the development and progression of various human diseases. Chapters detail miRNA biogenesis, isolating RNA, extracellular vesicles (EVs), circulating miRNAs, analyzing miRNA and miRDeep-P2, protocols for total RNA isolation from cells, cell-derived products, isolation and characterization of exosomes, serum, plasma specimens, and software tools. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, MicroRNA Profiling: Methods and Protocols, Second Edition aims to provide comprehensive and accessible methods to undergraduate, graduate, and established scientist.
This edited book, "Nucleic Acids - From Basic Aspects to Laboratory Tools", contains a series of chapters that highlight the development and status of the various aspects of the nucleic acids related to DNA chemistry and biology and the molecular application of these small DNA molecules and related synthetic analogues within biological systems. Furthermore, it is hoped that the information in the present book will be of value to those directly engaged in the handling and use of nucleic acids, and that this book will continue to meet the expectations and needs of all who are interested in the different fascinating aspects of molecular biology.
This book provides a practical and clinically oriented guide to the concepts of pathologic diagnosis of surgical specimens. Concise and highly illustrated chapters cover essential information required within patient management. Telepathology, ancillary techniques, and surgical oncology concepts are also examined. Surgical Pathology: A Practical Guide aims to bridge the knowledge gap between surgeons and pathologists to promote mutual understanding and a better working relationship. This book is relevant to general surgeons, and surgical oncologists, whether in training or in practice. It also serves as an introduction for first-year pathology residents, and medical students interested in surgical pathology.