Immunocytochemistry and In Situ Hybridization in the Biomedical Sciences

Immunocytochemistry and In Situ Hybridization in the Biomedical Sciences PDF Author: Julian E. Beesley
Publisher: Springer Science & Business Media
ISBN: 146120139X
Category : Science
Languages : en
Pages : 277

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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.

Immunocytochemistry and In Situ Hybridization in the Biomedical Sciences

Immunocytochemistry and In Situ Hybridization in the Biomedical Sciences PDF Author: Julian E. Beesley
Publisher: Springer Science & Business Media
ISBN: 146120139X
Category : Science
Languages : en
Pages : 277

Get Book Here

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.

Immunocytochemistry and in Situ Hybridization in the Biomedical Sciences

Immunocytochemistry and in Situ Hybridization in the Biomedical Sciences PDF Author:
Publisher:
ISBN:
Category : Immunocytochemistry
Languages : en
Pages : 0

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Book Description


In Situ Molecular Pathology and Co-Expression Analyses

In Situ Molecular Pathology and Co-Expression Analyses PDF Author: Gerard J. Nuovo
Publisher: Academic Press
ISBN: 0124160018
Category : Science
Languages : en
Pages : 298

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Book Description
In Situ Molecular Pathology and Co-Expression Analyses explains, in easy-to-understand language, simplified ways of understanding and performing in situ hybridization and immunohistochemistry tests. The book also focuses on straightforward protocols used to simultaneously detect two or more proteins/nucleic acids within intact tissue by doing co-expression analyses. The fields of in situ hybridization and immunohistochemistry have expanded rapidly due to the use of computer-based analysis. To get the most out of these automated platforms, researchers and diagnostic biomedical investigators must have a solid understanding of the basics of in situ-based tests, protocols, and regimens for troubleshooting. Practicing molecular pathologists, clinical chemists, and toxicologists, as well as clinicians and researchers in training, will benefit from this book's clear presentation of protocols and theoretical framework. - Includes over 200 easy-to-follow experimental protocols - Features chapter-ending summaries of "Key Points to Remember" to bring beginners up to speed with any seasoned veteran in the field - Offers two chapters written by industry leaders in the fields of in situ hybridization, immunohistochemistry, and computer software for co-expression analyses

In Situ Hybridization Methods

In Situ Hybridization Methods PDF Author: Giselbert Hauptmann
Publisher: Humana
ISBN: 9781493944637
Category : Medical
Languages : en
Pages : 0

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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.

Immunocytochemistry

Immunocytochemistry PDF Author: Richard W. Burry
Publisher: Springer Science & Business Media
ISBN: 1441913041
Category : Science
Languages : en
Pages : 225

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Book Description
Description: In biomedical research, because of a dramatic increase in productivity, immunocytochemistry has emerged as a major technique. The proposed book will provide the first practical guide to planning, performing, and evaluating immunocytochemical experiments. In today’s graduate education the emphasis is on doing research and not on formal class work. Graduate students therefore lack the background in many essential techniques necessary to perform research in fields in which they were not trained. As director of a university core microscopy facility which sees students and faculty from dozens of laboratories each year, Dr. Burry has surmised the vast majority of these novice microscope users need considerable help. In an attempt to educate users, Dr. Burry has initiated immunocytochemistry seminars and workshops which serve to train people in this powerful research tool. The proposed book is an outgrowth of these presentations and conversations with, by now, hundreds of people who have asked for help. The philosophy which separates this book from other books in this field is that it is practical, rather than academic. In looking at other important immunocytochemistry titles, the predominant orientation is academic, with the author attempting to comprehensively discuss the topic. For example, one book with sample preparation lists ten fixatives which can be used; however, only two such fixatives are commonly used today. In this particular title, the detailed discussion of old methods might be seen as important in establishing the author as an expert. By contrast, the approach for Burry’s book would be to discuss methods based on what works in animal research laboratories today, and focus only on the most productive methods. An additional distinction with this proposed book is the focus on animal research and not human pathology. There is a certification program for pathology technicians which requires them to learn a set body of material based on processing human tissue for examination by a pathologist. Many of the books on immunocytochemistry aim at this large pathology user base. Due to historical reasons, pathology laboratories process human tissues in a specific way and embed the tissue in paraffin, as has been done for over a century. In the last ten years, the power of immunocytochemistry in clinical diagnosis has become clear and has accordingly been adapted to pathology. However, the extensive processing needed for paraffin sections is not needed if the tissues are from research animals. Processing for animal-based tissues takes about a third of the time and results in higher quality images. The focus of this book is on processing these animal research tissues for immunocytochemistry. Today, there are no technique books which are aimed at this user base. As a subject matter expert in the area of the proposed book, Dr. Burry will make recommendations and offer opinions. Because this field is new and is emerging, there are numerous advantages of specific methods over other, more generalized methods. The purpose of this book is to show a novice how to do immunocytochemistry without engaging in a discussion of possible advanced methods. For the advanced user, there are several good books which discuss the unusual methods, yet for the novice there are currently none. Main Author : Richard W. Burry, The Ohio State University (United States). The Outline of the Book : Each chapter supplies a set of important principals and steps necessary for good immunocytochemistry. The information is distilled down to include only the most important points and does not attempt to cover infrequently used procedures or reagents. At the end of most chapters is a section on trouble-shooting many of the common problems using the Sherlock Holmes method. Each chapter also includes specific protocols which can be used. The goal of each chapter is to present the reader with enough information to successfully design experiments and solve many of the problems one may encounter. Using immunocytochemical protocols without the understanding of their workings is not advised, as the user will need to evaluate his or her results to determine whether the results are reliable. Such evaluation is extremely important for users who need reliable images which will clearly answer important scientific questions. 1. Introduction Definitions (immunocytochemistry and immunohistochemistry) Scope: animal research and not human pathology, paraffin sections, epitope retrieval, or immunohistochemistry Focus: fluorescence and enzyme detection Why do immunocytochemistry? Immunocytochemistry "individual study" rather than "population study" Example of a two-label experiment What is included in these chapters? Overview of the theory Background with enough information to help solve common problems. Advantages and disadvantages of different options Opinions and suggestions 2. Fixation and Sectioning Chemistry of fixation Denaturing vs cross-linking fixatives Application of fixative Perfusion, drop-in, cultures, fresh-frozen Selection of sample section type Sectioning tissue Rapid freezing, cryostat, freezing microtome, vibratome Storage of tissue Protocols 3. Antibodies Introduction Isoforms, structure, reactivity Generation Polyclonal vs monoclonal Antibodies as reagents Antibody specificity and sources Storage and handling 4. Labels for antibodies Fluorescence, enzymes and particulates Fluorescence theory Fluorescent labels - four generations Enzymes theory Selecting enzymes vs. fluorescence Selecting a label- advantages and disadvantages Protocols 5. Methods of applying antibodies Direct method Indirect method Antibody amplification methods ABC TSA Protocols 6. Blocking and Permeability Theory of blocking Theory of detergents Protocols 7. Procedure- Single primary antibody Planning steps Sample, fixation, sectioning Vehicle Antibody dilutions Controls Protocols 8. Multiple primary antibodies - primary antibodies of different species Procedure Controls Protocols 9. Multiple primary antibodies-primary antibodies of same species Block-between Zenon HRP-chromogen development High-titer incubations Controls Protocols 10. Microscopy Wide-field fluorescence microscope Confocal microscope Bright field—enzyme chromogen Choice Problems 11. Images Size, intensity, and pixels Manipulation—what is ethical? Manuscript Figures 11. Planning and Troubleshooting Scheme for discussion-making in planning experiments Case studies with Sherlock Holmes detective work 12. So you want to do electron microscopic ICC? Criteria in decision-making Summary of the two techniques

Morphology Methods

Morphology Methods PDF Author: Ricardo V. Lloyd
Publisher: Springer Science & Business Media
ISBN:
Category : Medical
Languages : en
Pages : 456

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Book Description
This text explores the most significant aspects of sophisticated and recent cell biological techniques that are needed to keep up with research in morphology. Researchers in morphology will find many step-by-step protocols to conduct their work including state-of -- the -- art laser capture microdissection methods, real time PCR and advanced FISH techniques. The chapters include detailed protocols for setting up and performing these techniques. This book covers all of the major areas relevant to molecular morphology today. Potential pitfalls and anticipated problems are also discussed. Morphology Methods will prove essential to researchers interested in molecular morphology for the purpose of experiments, and for teaching graduate, or professional students wishing to keep up with the literature.

Molecular Diagnostics

Molecular Diagnostics PDF Author: Wayne W. Grody
Publisher: Academic Press
ISBN: 0080919049
Category : Medical
Languages : en
Pages : 518

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Book Description
Advances in genomic and proteomic profiling of disease have transformed the field of molecular diagnostics, thus leading the way for a major revolution in clinical practice. While the range of tests for disease detection and staging is rapidly expanding, many physicians lack the knowledge required to determine which tests to order and how to interpret results. Molecular Diagnostics provides a complete guide to the use and interpretation of molecular testing in the clinical arena. No other available resource offers this emphasis, comprehensive scope, and practical utility in the clinical setting. - Serves as the definitivereference for molecular pathologists worldwide - Covers a variety of molecular techniques including next generation sequencing, tumor somatic cell genotyping, infectious and genetic disease tecting, and pharmacogenetics - Discusses in the detail issues concerning quality assurance, regulation, ethics, and future directions for the science

Biological Electron Microscopy

Biological Electron Microscopy PDF Author: Michael J. Dykstra
Publisher: Springer Science & Business Media
ISBN: 9780306477492
Category : Science
Languages : en
Pages : 558

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Book Description
Electron microscopy is frequently portrayed as a discipline that stands alone, separated from molecular biology, light microscopy, physiology, and biochemistry, among other disciplines. It is also presented as a technically demanding discipline operating largely in the sphere of "black boxes" and governed by many absolute laws of procedure. At the introductory level, this portrayal does the discipline and the student a disservice. The instrumentation we use is complex, but ultimately understandable and, more importantly, repairable. The procedures we employ for preparing tissues and cells are not totally understood, but enough information is available to allow investigators to make reasonable choices concerning the best techniques to apply to their parti cular problems. There are countless specialized techniques in the field of electron and light microscopy that require the acquisition of specialized knowledge, particularly for interpretation of results (electron tomography and energy dispersive spectroscopy immediately come to mind), but most laboratories possessing the equipment to effect these approaches have specialists to help the casual user. The advent of computer operated electron microscopes has also broadened access to these instruments, allowing users with little technical knowledge about electron microscope design to quickly become operators. This has been a welcome advance, because earlier instru ments required a level of knowledge about electron optics and vacuum systems to produce optimal photographs and to avoid "crashing" the instruments that typically made it difficult for beginners.

In Situ Hybridization in Neurobiology

In Situ Hybridization in Neurobiology PDF Author: James H. Eberwine
Publisher:
ISBN: 9780195075076
Category : Medical
Languages : en
Pages : 230

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Book Description
In situ hybridization has become an important and widely used research tool for neurobiologists since the authors' first book on the subject, In Situ Hybridization: Applications to Neurobiology (Oxford, 1987). This new volume examines the methodological advances that have been made in in situ hybridization techniques and their impact on the study of the nervous system. These include methods for improving sensitivity, ways of examining multiple mRNAs within the same section, ways of examining both proteins and mRNAs within the same section, and methods for using in situ hybridization technology to examine areas of cellular biological significance, such as translational control. The development of alternative labels to radioactivity for the detection of hybridized probes, and the combined use of in situ hybridization and immunocytochemistry is also covered in this book. It discusses the quantification of in situ hybridization signal and presents an overview of information generated by using in situ hybridization in diagnostic medicine. This volume will be essential to researchers in the neurosciences as well as those in other disciplines.

Essential Concepts in Molecular Pathology

Essential Concepts in Molecular Pathology PDF Author: William B. Coleman
Publisher: Academic Press
ISBN: 0128132582
Category : Medical
Languages : en
Pages : 636

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Book Description
Essential Concepts in Molecular Pathology, Second Edition, offers an introduction to molecular genetics and the "molecular" aspects of human disease. The book illustrates how pathologists harness their understanding of these entities to develop new diagnostics and treatments for various human diseases. This new edition offers pathology, genetics residents, and molecular pathology fellows an advanced understanding of the molecular mechanisms of disease that goes beyond what they learned in medical and graduate school. By bridging molecular concepts of pathogenesis to the clinical expression of disease in cell, tissue and organ, this fully updated, introductory reference provides the background necessary for an understanding of today's advances in pathology and medicine. - Explains the practice of "molecular medicine" and the translational aspects of molecular pathology, including molecular diagnostics, molecular assessment and personalized medicine - Orients non-pathologists on what pathologists look for and how they interpret their observational findings based on histopathology - Provides the reader with what is missing from most targeted introductions to pathology—the cell biology behind pathophysiology