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Author: Luis Del Valle
Publisher: Humana
ISBN: 9781071619506
Category : Medical
Languages : en
Pages : 0
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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 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
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
Author: Julia M. Polak
Publisher: Springer
ISBN: 9780387915135
Category : Immunocytochemistry
Languages : en
Pages : 141
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Book Description
Handbook describing practical immunocytochemical techniques and their underlying rationales. Discusses the problems of specificity and sensitivity.
Author: 0 J.M. Polak and S. Van Noorden
Publisher: Taylor & Francis
ISBN: 1000947947
Category : Science
Languages : en
Pages : 170
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Book Description
This book provides a practically directed basis for carrying out immunocytochemical methods with enough theoretical information to allow a newcomer to understand the whys and wherefores of the technique. It explains tissue preparation, cytological preparations and new methods of multiple staining.
Author: Julia M. Polak
Publisher: Oxford University Press, USA
ISBN:
Category : Architecture
Languages : en
Pages : 76
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Book Description
This work gives a concise exposition of the different immunocytochemical methods for identifying tissue constituents. It discusses ways of preparing tissues according to the antigen to be localized, and most commonly used methods, sources of error and nonspecifity, and techniques for recording results photographically.
Author: Luis Del Valle
Publisher: Humana
ISBN: 9781071619476
Category : Medical
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 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
Author: Fan Lin
Publisher: Springer
ISBN: 1493915789
Category : Medical
Languages : en
Pages : 756
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Book Description
In a conceptually current, quick-reference, Question & Answer format, the second edition of Handbook of Practical Immunohistochemistry: Frequently Asked Questions continues to provide a comprehensive and yet concise state-of-the-art overview of the major issues specific to the field of immunohistochemistry. With links to the authors Immunohistochemical Laboratory website, this volume creates a current and up-to-date information system on immunohistochemistry. This includes access to tissue microarrays (TMA) of over 10,000 tumors and normal tissue to validate common diagnostic panels and provide the best reproducible data for diagnostic purposes. Fully revised and updated from the first edition, the new features of the second edition include over 200 additional questions or revised questions with an IHC panel to answer each question; over 250 new color photos and illustrations; over 20 new useful biomarkers; hundreds of new references; several new chapters to cover phosphoproteins, rabbit monoclonal antibodies, multiplex IHC stains, overview of predictive biomarkers, and integration of IHC into molecular pathology; many new coauthors who are international experts in a related field; many updated IHC panels using Geisinger IHC data collected from over 10,000 tumors and normal tissues; and updated appendices containing detailed antibody information for both manual and automated staining procedures. Comprehensive yet practical and concise, the Handbook of Practical Immunohistochemistry: Frequently Asked Questions, Second Edition will be of great value for surgical pathologists, pathology residents and fellows, cytopathologists, and cytotechnologists.
Author: Gareth Griffiths
Publisher: Springer Science & Business Media
ISBN: 3642770959
Category : Science
Languages : en
Pages : 477
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Book Description
Electron microscopy in the biological sciences can be divided into two disciplines. The first, concerned with high resolution detail of particles or periodic structures, is mostly based on sound theoretical principles of physics. The second, by far the larger discipline, is interested in the information obtainable from thin sections. The theoretical back ground to those groups of techniques for preparing and looking at thin sections is often inexact and "loose", for want of a better word. What should be chemistry is often closer to alchemy. This kind of electron microscopy is often enshrined with mystical recipes, handed down from generation to generation. Admittedly, many of the processes involved, such as those required to embed tissue in epoxy resins, involve multiple interconnected steps, which make it difficult to follow the details of anyone of these steps. If all these steps are shrouded in some mystery, however, can one really trust the final image that emerges on the EM screen? When we present the data in some semi quantitative form is there really no better way to do it than to categorize the parameters with ++, +/-, etc? What happens when one labels the sections with antibodies? Does the whole business necess arily need to be more of an "art" than a "science"? Upon reflecting on these problems in 1981, I had the impression that many of the multi-authored textbooks that existed then (and that have appeared since) tended to exacerbate or at least perpetuate this
Author: Julia M. Polak
Publisher: Springer
ISBN: 9789813083356
Category : Immunocytochemistry
Languages : en
Pages : 141
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Book Description
Author: Mai P. Hoang
Publisher: Cambridge University Press
ISBN: 1107150167
Category : Medical
Languages : en
Pages : 315
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Book Description
One of only a few textbooks on the market dedicated to the important role of immunohistochemistry in diagnostic dermatopathology.
Author: Constance Oliver
Publisher: Humana Press
ISBN: 9781617796821
Category : Medical
Languages : en
Pages : 416
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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.
