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.

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.

Histochemistry deals with the activities of chemical components in cells, and immunohistochemistry addresses the function of cell types in tissue or organs, such as those leading to acceptance or rejection of grafts or organs. This book is a methods volume focusing on antigen retrieval, particularly methods used in disease-related antigens. Because the book is a methods volume and a lab manual, it will have an audience of pathologists, biochemists, and lab technicians.

1. Introduction (S Renshaw) 2. Antibodies for immunochemistry (C Onley) 3. The selection of reporter labels (S Mardle) 4. Immunochemical staining techniques (S Renshaw) 5. Multiple immunochemical staining techniques (I Jones) 6. Confocal microscopy and immunohistochemistry (M Cuttle) 7. Ultrastructural immunochemistry (J Skepper) 8. Image capture, analysis and quantification (D Tannahill) 9. Quality assurance in immunohistochemistry (P Jackson) 10. Automated immunochemistry (E Schenck) List of suppliers; Index

Classical histology has been augmented by immunohistochemistry (the use of specific antibodies to stain particular molecular species in situ). Immunohistochemistry has allowed the identification of many more cell types than could be visualized by classical histology, particularly in the immune system and among the scattered hormone-secreting cells of the endocrine system. This book discusses all aspects of immunohistochemistry and in situ hybridization technologies and the important role they play in reaching a cancer diagnosis. It provides step-by-step instructions on the methods of additional molecular technologies such as DNA microarrays, and microdissection, along with the benefits and limitations of each method. * The only book available that translates molecular genetics into cancer diagnosis * Methods were developed by internationally-recognized experts and presented in step-by-step manner * Results of each Immunohistochemical and in situ hybridization are presented in the form of color illustrations

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.

The past several decades have witnessed an impressive array of conceptual and techno logical advances in the biomedical sciences. Much of the progress in this area has developed directly as a result of new morphology-based methods that have permitted the assessment of chemical, enzymatic, immunological, and molecular parameters at the cellular and tissue levels. Additional novel approaches including laser capture microdissection have also emerged for the acquisition of homogeneous cell popula tions for molecular analyses. These methodologies have literally reshaped the approaches to fundamental biological questions and have also had a major impact in the area of diagnostic pathology. Much of the groundwork for the development of morphological methods was estab th lished in the early part of the 19 century by Francois-Vincent Raspail, generally acknowledged as the founder of the science of histochemistry. The earliest work in the field was primarily in the hands of botanists and many of the approaches to the under standing of the chemical composition of cells and tissues involved techniques such as microincineration, which destroyed structural integrity. The development of aniline th dyes in the early 20 century served as a major impetus to studies of the structural rather than chemical composition of tissue. Later in the century, however, the focus returned to the identification of chemical constituents in the context of intact cell and tissue structure.