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Immunohistochemistry and Immunocytochemistry by Luis Del Valle Pdf
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
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 and Immunocytochemistry by Simon Renshaw Pdf
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.
Fine Structure Immunocytochemistry by Gareth Griffiths Pdf
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
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.
Resin Microscopy and On-Section Immunocytochemistry by Geoffrey R. Newman,Jan A. Hobot Pdf
Since antibodies tagged with markers have been developed, immunocytochemistry has become an important method for identifying tissue substances and the localisation of nucleic acid in tissue by in situ hybridisation in molecular biology. This laboratory book covers the embedding of tissue using less sensitive epoxy resin methods to the more sensitive procedures employing the acrylics. The possibilities are discussed and results are presented so that an understanding of the techniques can be acquired.
Immunocytochemistry for Steroid Receptors by Louis P. Pertschuk Pdf
A comprehensive review of the literature on steroid hormone receptor immuncytochemistry. It includes the development of the first monoclonal antireceptor antibody and initial procedures designed to localize steroid receptors in situ up to the present. The step-by-step instructions provide easy reading of topics such as tissue handling, processing, and interpretation. This text places particular emphasis on the studies of human material-especially mammary carcinoma. It presents specific details with chapters on estrogen and progesterone receptors in breast malignancies and benign breast disease. Each section of this outstanding book contains comprehensive photomicrographs that illustrate the most salient features discussed. Detailed graphs pertaining to theoretical particulars concerning technology and the relationship between image analysis and endocrine response are also included. This monograph is an excellent resource for all researchers concerned with hormone-dependent tissues, pathologists who have an interest in working with novel techniques for localizing steroid hormone receptors, and oncologists who must use assay results in the treatment of breast cancer.
Color Atlas of Immunocytochemistry in Diagnostic Cytology by Parvin Ganjei-Azar,Mehrdad Nadji Pdf
This atlas examines the benefits and limitations of immunocytochemistry (ICC) in contemporary cytology. The text addresses offers practical advice on choice of markers, and illustrates the use of ICC in fine needle aspiration and body cavity fluid cytology. This practical handbook will allow for quick reference in the selection and interpretation of markers in specific differential diagnostic in the daily practice of diagnostic cytology. Includes 299 color illustrations.
Immunocytochemistry by Julia M. Polak,Susan Van Noorden Pdf
Immunocytochemistry: Practical Applications in Pathology and Biology focuses on the processes, principles, methodologies, and approaches involved in the applications of immunocytochemistry in pathology and biology. The selection first takes a look at the development and scope of immunocytochemistry; techniques and practice used in immunocytochemistry; and raising and testing antibodies for immunocytochemistry. The book then ponders on raising antibodies to small peptides, use of semithin frozen sections in immunocytochemistry, and colloidal gold probes in immunocytochemistry. Discussions focus on the production of gold probes, localization of mouse serum albumin in the mammary gland, immunogens and the immune response, production of antisera, and antibody specificity in immunocytochemistry. The publication takes a look at double immunoenzymatic labelling, lectin histochemistry, immunocytochemical localization of noradrenaline, adrenaline, and serotonin, and immunocytochemistry of cell and tissue cultures. Immunocytochemistry in endocrine pathology and indirect immunofluorescence in the study and diagnosis of organ-specific autoimmune diseases are also discussed. The selection is a dependable source of information for researchers interested the practical applications of immunocytochemistry in pathology and biology.
Applications of Immunocytochemistry by Hesam Dehghani Pdf
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.
0 J.M. Polak and S. Van Noorden,Dame Julia Polak,Susan Van Noorden
Author : 0 J.M. Polak and S. Van Noorden,Dame Julia Polak,Susan Van Noorden Publisher : Taylor & Francis Page : 170 pages File Size : 45,8 Mb Release : 2023-04-28 Category : Science ISBN : 9781000947946
Introduction to Immunocytochemistry by 0 J.M. Polak and S. Van Noorden,Dame Julia Polak,Susan Van Noorden Pdf
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 of Plant Cells by Kevin Vaughn Pdf
Immunocytochemistry of plant cells is the first book exclusively dedicated to this topic. The first and largest portion of the book is concerned with a group of proven protocols and variations on these protocols that might prove useful, many developed or modified in the author's laboratory. The second portion of the book covers the studies that have been published previously on each of the plant organelles. Numerous state of the art micrographs from researchers around the world are included to demonstrate typical results.
Applications of Immunocytochemistry by Hesam Dehghani Pdf
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.
Practical Immunocytochemistry in Diagnostic Cytology by Eugenio Leonardo,Ricardo H. Bardales Pdf
This book provides current information for a judicious use of conventional and molecular ICC markers performed in cytologic samples to help in the cost-effective diagnosis of superficial and deep-seated masses. By providing both an eminently practical information for appropriate technical preparation of cytologic samples and a systematic exposure of the immunophenotypic and biomolecular characteristics of normal and pathological cells, the reported morphological-molecular findings highlight the characteristics of different lesions that are useful for the practical solution of a particular differential diagnosis. Chapters include description of the cytomorphology followed by a practical ICC staining pattern for each entity. Cytologic materials include serous effusions, exfoliative cytology, fine needle aspiration cytology of superficial and deep-seated masses, and squash preparation from central nervous system masses. Practical Immunocytochemistry in Diagnostic Cytology serves as a valuable resource towards understanding critical differences between tests performed on tissue samples and those carried out on cytologic samples.
Immunocytochemistry in Diagnostic Cytology by Pranab Dey Pdf
Cytology refers to a branch of pathology that deals with making diagnoses of diseases and conditions through the examination of tissue samples from the body (MedicineNet.com). Immunocytochemistry is a laboratory method that uses antibodies to check for certain antigens (markers) in a sample of cells. The antibodies are usually linked to an enzyme or a fluorescent dye. After the antibodies bind to the antigen in the cell sample, the enzyme or dye is activated, and the antigen can then be seen under a microscope. Immunocytochemistry is used to help diagnose diseases, such as cancer. It may also be used to help tell the difference between different types of cancer (National Cancer Institute). This book is a comprehensive guide to the techniques and application of immunocytochemistry in cytology. Divided into two parts, the first section discusses basic principles and preparation, techniques and quality control, and automated immune staining. Section Two covers diagnostic applications of immunocytochemistry for many different types of tumour. The text is highly illustrated with microphotographs, tables and boxes to assist learning and interpretation of findings for accurate diagnosis. Key points Comprehensive guide to techniques and application of immunocytochemistry in cytology Provides thorough understanding of basic principles and methods Covers diagnostic applications for many different types of tumour Highly illustrated with microphotographs, tables and boxes