Electron Microscopic Immunocytochemistry

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Fine Structure Immunocytochemistry

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Author : Gareth Griffiths
language : en
Publisher: Springer Science & Business Media
Release Date : 2012-12-06

Fine Structure Immunocytochemistry written by Gareth Griffiths 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 2012-12-06 with Science categories.

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

Electron Microscopic Immunocytochemistry

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Author : Julia M. Polak
language : en
Publisher:
Release Date : 1992

Electron Microscopic Immunocytochemistry written by Julia M. Polak and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1992 with Medical categories.

Electron microscopic immunocytochemistry is a powerful technique combining the resolution of the electron microscope with the specificity and sensitivity of immunocytochemistry. This book discusses methods designed to make this intricate procedure more accessible, and covers recent developments that have simplified the technique and broadened the range of applications. It presents the principles of EM immunocytochemistry, detailed protocols, and examples of ways in which the technique can be applied to a variety of fields, including biomedical research and diagnostic pathology. The book begins with an overview of EM immunocytochemical procedures and of applications in histopathology. The three major chapters cover post-embedding, pre-embedding, and ultra-thin cryosection techniques. The final section illustrates applications of EM immunocytochemistry to the study of endocrine tumors, renal disease, neuropathology, dermatology, and microbiology. The emphasis throughout this detailed volume is on providing practical guidance to scientists and clinicians who may be using the technique for the first time.

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

Electron Microscopic Cytochemistry And Immunocytochemistry In Biomedicine

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Author : Kazuo Ogawa
language : en
Publisher: CRC Press
Release Date : 1993

Electron Microscopic Cytochemistry And Immunocytochemistry In Biomedicine written by Kazuo Ogawa and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 1993 with Medical categories.

Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine provides up-to-date, comprehensive surveys and descriptions of electron microscopic cytochemical and immunocytochemical methods. It also reviews selected topics of cell biology, pathology, histology, and biomedicine. These reviews illustrate the fundamental contributions of electron microscopic cytochemistry and immunocytochemistry to biology and pathology. By bringing together methodology and the application of methods, this book presents a broad discussion of the contributions, potentials, and limitations of cytochemistry and immunocytochemistry. Electron Microscopic Cytochemistry and Immunocytochemistry in Biomedicine will be a useful reference volume for cell biologists, histologists, anatomists, pathologists, physiologists, immunologists, and clinical researchers concerned with the basic mechanisms of diseases.

Immuno Gold Electron Microscopy In Virus Diagnosis And Research

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Author : Alexander D. Hyatt
language : en
Publisher: CRC Press
Release Date : 2024-12-20

Immuno Gold Electron Microscopy In Virus Diagnosis And Research written by Alexander D. Hyatt and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2024-12-20 with Science categories.

This book presents a wide variety of immuno-gold techniques for use in virus diagnosis and research. Protocols are presented for state-of-the-art techniques, including in situ hybridization, freeze substitution, and the utilization of ultra-small probes and replicas for use by virologists and electron microscopists identifying and studying viruses, their components, and replication in cells. The procedures are described by eminent scientists and are pertinent to both experienced researchers and newcomers to this field who are interested in the localization of low antigenic mass structures.

Introduction To Immunocytochemistry

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Author : 0 J.M. Polak and S. Van Noorden
language : en
Publisher: Taylor & Francis
Release Date : 2023-04-28

Introduction To Immunocytochemistry written by 0 J.M. Polak and S. Van Noorden and has been published by Taylor & Francis this book supported file pdf, txt, epub, kindle and other format this book has been release on 2023-04-28 with Medical categories.

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.

Immunocytochemistry

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Author : Lars-Inge Larsson
language : en
Publisher: CRC Press
Release Date : 2020-11-25

Immunocytochemistry written by Lars-Inge Larsson and has been published by CRC Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2020-11-25 with Medical categories.

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.

Biomedical Electron Microscopy

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Author : Arvid B. Maunsbach
language : en
Publisher: Elsevier
Release Date : 1998-11-03

Biomedical Electron Microscopy written by Arvid B. Maunsbach and has been published by Elsevier this book supported file pdf, txt, epub, kindle and other format this book has been release on 1998-11-03 with Science categories.

This comprehensive reference illustrates optimal preparation methods in biological electron microscopy compared with common methodological problems. Not only will the basic methodologies of transmission electron microscopy like fixation, microtomy, and microscopy be presented, but the authors also endeavor to illustrate more specialized techniques such as negative staining, autoradiography, cytochemistry, immunoelectron microscopy, and computer-assisted image analysis. - Authored by the key leaders in the biological electron microscopy field - Illustrates both optimal and suboptimal or artifactual results in a variety of electron microscopy disciplines - Introduces students on how to read and interpret electron micrographs

Diagnostic Immunocytochemistry And Electron Microscopy

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Author : Hossein M. Yazdi
language : en
Publisher: Igaku-Shoin Medical Publishers
Release Date : 1992

Diagnostic Immunocytochemistry And Electron Microscopy written by Hossein M. Yazdi and has been published by Igaku-Shoin Medical Publishers this book supported file pdf, txt, epub, kindle and other format this book has been release on 1992 with Medical categories.

Electron Microscopy Of Model Systems

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Author :
language : en
Publisher: Academic Press
Release Date : 2010-09-24

Electron Microscopy Of Model Systems written by and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2010-09-24 with Science categories.

The volume covers the preparation and analysis of model systems for biological electron microscopy. The volume has chapters about prokaryotic as well as eukaryotic systems that are used as so-called model organisms in modern cell biology. These systems include the most popular systems, such as budding and fission yeast, the roundworm C. elegans, the fly Drosophila, zebrafish, mouse, and Arabidopsis, but also organisms that are less frequently used in cell biology, such as Chlamydomonas, Dictyostelium, Trypanosoma, faltworms, Axolotl and others. In addition, tissues and tissue culture systems are also covered. These systems are used for very diverse areas of cell biology, such as cell division, abscission, intracellular transport, cytoskeletal organization, tissue regeneration and others. Moreover, this issue presents the currently most important methods for the preparation of biological specimens. This volume, however, is not a classic EM methods book. The methods are not the main focus of this issue. The main goal here is to cover the methods in the context of the specific requirements of specimen preparation for each model organism or systems. This will be the first compendium covering the various aspects of sample preparation of very diverse biological systems. - Covers the preparation and analysis of model systems for biological electron microscopy - Includes the most popular systems but also organisms that are less frequently used in cell biology - Presents the currently most important methods for the preparation of biological specimens - First compendium covering the various aspects of sample preparation of very diverse biological systems