Immunocytochemical Methods and Protocols

Immunocytochemical Methods and Protocols PDF Author: Constance Oliver
Publisher: Humana Press
ISBN: 9781588294630
Category : Medical
Languages : en
Pages : 0

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Book Description
Antibodies tagged with fuorescent markers have been used in histochemistry for over 50 years. Although early applications were focused on the detection of microbial antigens in tissues, the use of immunocytochemical methods now has spread to include the det- tion of a wide array of antigens including proteins, carbohydrates, and lipids from virtually any organism. Today, immunohistochemistry is widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions. The method gains its strength from the extremely sensitive interaction of a specifc antibody with its antigen. For some scientifc areas, books have been published on applications of immu- cytochemical techniques specifc to that area. What distinguished Immunocytochemical Methods and Protocols from earlier books when it was frst published was its broad appeal to investigators across all disciplines, including those in both research and clinical settings. The methods and protocols p- sented in the frst edition were designed to be general in their application; the accompa- ing “Notes” provided the reader with invaluable assistance in adapting or troubleshooting the protocols. These strengths continued to hold true for the second edition and again for the third edition. Since the publication of the frst edition, the application of immuno- tochemical techniques in the clinical laboratory has continued to rise and this third edition provides methods that are applicable to basic research as well as to the clinical laboratory.

Immunocytochemical Methods and Protocols

Immunocytochemical Methods and Protocols PDF Author: Constance Oliver
Publisher: Humana Press
ISBN: 9781588294630
Category : Medical
Languages : en
Pages : 0

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Book Description
Antibodies tagged with fuorescent markers have been used in histochemistry for over 50 years. Although early applications were focused on the detection of microbial antigens in tissues, the use of immunocytochemical methods now has spread to include the det- tion of a wide array of antigens including proteins, carbohydrates, and lipids from virtually any organism. Today, immunohistochemistry is widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions. The method gains its strength from the extremely sensitive interaction of a specifc antibody with its antigen. For some scientifc areas, books have been published on applications of immu- cytochemical techniques specifc to that area. What distinguished Immunocytochemical Methods and Protocols from earlier books when it was frst published was its broad appeal to investigators across all disciplines, including those in both research and clinical settings. The methods and protocols p- sented in the frst edition were designed to be general in their application; the accompa- ing “Notes” provided the reader with invaluable assistance in adapting or troubleshooting the protocols. These strengths continued to hold true for the second edition and again for the third edition. Since the publication of the frst edition, the application of immuno- tochemical techniques in the clinical laboratory has continued to rise and this third edition provides methods that are applicable to basic research as well as to the clinical laboratory.

Immunohistochemistry and Immunocytochemistry

Immunohistochemistry and Immunocytochemistry PDF Author: Luis Del Valle
Publisher:
ISBN: 9781071619483
Category : Electronic books
Languages : en
Pages : 286

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Book Description
This volume provides a comprehensive reference guide for researchers to study the applications of labeled antibodies. Chapters guide reader through the the and practice of immunohistochemistry, immunocytochemistry and immunofluorescence techniques. 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 useful tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Immunohistochemistry and Immunofluorescence: Methods and Protocols aims to be a useful practical guide to scientists to help further their study in this field.

Neuroanatomical Tract-Tracing Methods 2

Neuroanatomical Tract-Tracing Methods 2 PDF Author: Laszlo Zaborszky
Publisher: Springer Science & Business Media
ISBN: 1475720556
Category : Medical
Languages : en
Pages : 429

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Book Description
This new edition presents readers with the latest information on neuroscience. This book explores the advances in molecular techniques, genomics and proteomics and the progress in fluorescence.

Immunocytochemical Methods and Protocols

Immunocytochemical Methods and Protocols PDF Author: Lorette C. Javois
Publisher: Springer Science & Business Media
ISBN: 1592595219
Category : Medical
Languages : en
Pages : 414

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Book Description
The principle that antibodies can be used as cytochemical agents provided they are tagged with suitable markers has been evident for over 50 years. During this time the use of immunocytochemical meth ods has spread to a wide array of biological disciplines. Early applica tions focused on the detection of microbial antigens in tissues, while more recent applications have used monoclonal antibodies to study cell differentiation during embryonic development. For a select few disci plines, volumes have been published focusing on the specific applica tion of immunocytochemical techniques to that discipline. What distinguishes the present book, Immunocytochemical Meth ods and Protocols, from earlier books is its broad appeal to researchers in all disciplines, including those in both research and clinical settings. The methods and protocols presented here are designed to be general in their application and the accompanying "Notes" provide invaluable assistance in adapting or troubleshooting the protocols. Interspersed throughout the book are chapters providing overviews of select topics related to immunocytochemistry.

Immunohistochemistry and Immunocytochemistry

Immunohistochemistry and Immunocytochemistry PDF Author: Simon Renshaw
Publisher: John Wiley & Sons
ISBN: 1118717775
Category : Science
Languages : en
Pages : 258

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Book Description
Immunohistochemistry and immunocytochemistry are invaluable tools for the visualization of tissue and cellular antigens in diagnostic and biological research environments. The need to obtain accurate, reliable and reproducible results is paramount. It is with this fundamental aim in mind that we have compiled Immunohistochemistry: Essential Methods. We have achieved this by examining each aspect of immunochemistry in turn, with each chapter including detailed information regarding the subject matter in question. Each chapter is written by an expert in their field and includes protocols that are typically used in their own research. Subjects covered are, amongst others, antibodies and their production; selection of reporter labels; immunochemical staining methods and experimental design (both using single and multiple reporter labels); quality assurance; automated immunochemistry; confocal microscopy and electron microscopy. In addition, benefits and limitations of each approach are discussed within the chapters.

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

Immunohistochemistry: Basics and Methods

Immunohistochemistry: Basics and Methods PDF Author: Igor B. Buchwalow
Publisher: Springer Science & Business Media
ISBN: 3642046096
Category : Medical
Languages : en
Pages : 153

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Book Description
This concise yet comprehensive guide to the methods and protocols of immunohistochemistry covers established techniques and current developments in the field such as the use of epitope tags, multiple immunolabeling and diagnostic immunohistochemistry.

Applications of Immunocytochemistry

Applications of Immunocytochemistry PDF Author: Hesam Dehghani
Publisher: IntechOpen
ISBN: 9789535102298
Category : Medical
Languages : en
Pages : 332

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Book Description
Immunocytochemistry is classically defined as a procedure to detect antigens in cellular contexts using antibodies. However, over the years many aspects of this procedure have evolved within a plethora of experimental setups. There are different ways to prepare a given specimen, different kinds of antibodies to apply, different techniques for imaging, and different methods of analyzing the data. In this book, various ways of performing each individual step of immunocytochemistry in different cellular contexts are exemplified and discussed. Applications of Immunocytochemistry offers technical and background information on different steps of immunocytochemistry and presents the application of this technique and its adaptations in cell lines, neural tissue, pancreatic tissue, sputum cells, sperm cells, preimplantation embryo, arabidopsis, fish gonads, and Leishmania.

Immunocytochemistry

Immunocytochemistry PDF Author: Lars-Inge Larsson
Publisher: CRC Press
ISBN: 9780849360787
Category : Medical
Languages : en
Pages : 292

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Book Description
A complete and balanced overview of all aspects of immunocytochemistry is presented providing a clear understanding of their impact on experiment. All available techniques and many diagnostic and research applications are included, as well as practical step-by-step instructions for carrying out recommended methods. Intended for the novice as well as the experienced researchers.

Basic Methods in Antibody Production and Characterization

Basic Methods in Antibody Production and Characterization PDF Author: Gary C. Howard
Publisher: CRC Press
ISBN: 142003653X
Category : Medical
Languages : en
Pages : 286

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Book Description
Written for researchers and professionals in the fields of biomedical research, immunology, biochemistry, molecular biology, pathology, and biotechnology, Basic Methods in Antibody Production and Characterization uses a cookbook approach to presenting the methods for the production, characterization, and use of antibodies. Antibodies described