Immunocytochemical Methods And Protocols

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Immunocytochemical Methods And Protocols

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Author : Constance Oliver
language : en
Publisher: Humana Press
Release Date : 2012-04-06

Immunocytochemical Methods And Protocols written by Constance Oliver and has been published by Humana Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-04-06 with Medical categories.

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

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Author : Lorette C. Javois
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-02

Immunocytochemical Methods And Protocols written by Lorette C. Javois and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-02 with Medical categories.

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

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Author : Luis Del Valle
language : en
Publisher: Humana
Release Date : 2021-12-04

Immunohistochemistry And Immunocytochemistry written by Luis Del Valle and has been published by Humana this book supported file pdf, txt, epub, kindle and other format this book has been release on 2021-12-04 with Medical categories.

This volume provides a comprehensive reference guide for researchers to study the applications of labeled antibodies. Chapters guide reader through the theory 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

Molecular Biomethods Handbook

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Author : Ralph Rapley
language : en
Publisher: Springer
Release Date : 2007-10-09

Molecular Biomethods Handbook written by Ralph Rapley and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2007-10-09 with Science categories.

An authoritative team of investigators illuminate the core bioanalytical techniques used every day in their own laboratories, and laboratories throughout the world. These highly experienced scientists fully explain both the theory behind, and the application of, these key techniques, and include extensive references for those seeking detailed laboratory protocols. The techniques covered range from the extraction, separation, detection, and characterization of nucleic acids to gene cloning and library production, mapping, expression, transgenesis, differential display, and DNA profiling, to name a few. Numerous key protein methods, as well as support and related techniques, are also included. The goal is to provide established scientists and novices who are new to these techniques with a deeper understanding of the widest variety of biotechniques and their applications.

Immunohistochemistry Basics And Methods

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Author : Igor B. Buchwalow
language : en
Publisher: Springer Science & Business Media
Release Date : 2010-01-26

Immunohistochemistry Basics And Methods written by Igor B. Buchwalow and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010-01-26 with Medical categories.

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.

Neuroprotection Methods And Protocols

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Author : Tiziana Borsello
language : en
Publisher: Springer Science & Business Media
Release Date : 2007-08-08

Neuroprotection Methods And Protocols written by Tiziana Borsello and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2007-08-08 with Medical categories.

This book examines current research into the role of neuronal death in cell signaling pathways, and its role in neurodegenerative diseases, such as Alzheimer's and Parkinson's. After introducing neurodegenerative, traumatic, and ishemic disorders, the authors cover in vitro and animal systems, and cellular and molecular mechanisms.

Confocal Microscopy

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Author : Stephen W. Paddock
language : en
Publisher: Springer Science & Business Media
Release Date : 2008-02-03

Confocal Microscopy written by Stephen W. Paddock and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2008-02-03 with Science categories.

Immunohistochemistry

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Author : Alexander E. Kalyuzhny
language : en
Publisher: Springer
Release Date : 2016-03-24

Immunohistochemistry written by Alexander E. Kalyuzhny and has been published by Springer this book supported file pdf, txt, epub, kindle and other format this book has been release on 2016-03-24 with Science categories.

Aimed at both beginners and experienced researchers unfamiliar with the technique, this book provides a concise and easy-to-understand guide to using immunohistochemistry. It identifies the proper place and purpose of each component of immunohistochemistry, emphasizes the components that are of critical importance, and explains the logistics of experimental approaches. Understanding the principles of immunohistochemistry allows for the identification and localization of protein targets and is of crucial use in the following experiments: · producing transgenic animals · studying the effects of drug treatments · electrophysiological studies · animal surgery · isolating and differentiating stem cells · super-resolution microscopy Immunohistochemistry for the Non-Expert is aimed at a wide audience, including molecular biologists, pharmacologists, immunologists, chemists, cell biologists, physiologists, and researchers involved with animal anatomy studies.

Immunocytochemistry

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Author : Richard W. Burry
language : en
Publisher: Springer Science & Business Media
Release Date : 2009-12-08

Immunocytochemistry written by Richard W. Burry and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2009-12-08 with Science categories.

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