Immunocytochemistry and in situ hybridization are widely used biomedical sciences. They are essential in medical diagnosis and in cell biology research. Affinity labeling is the central goal of the experimental strategy involving a series of techniques in a logical order; from the effects of specimen fixation, through specimen preparation to expose the antigen, to optimizing immunolabeling, to assessing the result and finally to safety considerations. Numerous examples of these techniques in biomedical sciences are included, as well as experimental assays and practical tips. This survey of methods will serve as an invaluable reference source in any laboratory setting (academic, industrial or clinical) involved in research in almost every branch of biology or medicine, as well as in pharmaceutical, biotechnological and clinical applications.

Major advancements in the field of in situ molecular pathology have occurred since publication of the first edition. In Situ Molecular Pathology and Co-expression Analyses, Second Edition, continues to teach both the molecular basis for the improvements and the actual protocols. This is the unique feature that separates it from the pack of other "cook-book" type approaches. The fields of in situ hybridization and immunohistochemistry have expanded rapidly where computer-based analyses systems have greatly expanded the power of these methods. Further, knowledge of the marked improvements in the reagents themselves since the first edition can make the difference of excellent versus misleading data. The automated platforms require that researchers and diagnostic biomedical investigators have a good understanding of the basics of in situ based tests, protocols, and biochemistry for troubleshooting in order to maximize the use of these platforms. This second edition focuses attention on straightforward protocols used to simultaneously detect two or more proteins/nucleic acids within intact tissue by doing co-expression analyses. Practicing molecular pathologists, diagnostic pathologists, laboratory directors, and toxicologists, as well as clinicians and researchers in training, will benefit from this clear presentation of protocols and theoretical framework. Data derived from in situ hybridization and immunohistochemistry. Explains the theory and foundation of immunohistochemistry and in situ hybridization and presents easy-to-follow experimental protocols with tricks of the trade Includes two new chapters: Recent improvements in immunohistochemistry and in situ hybridization, Quality control for immunohistochemistry and in situ hybridization: How to know if the color change is signal or background The second edition also includes a detailed test to help one learn the basics of histologic interpretation of tissues and a separate detailed test in how to differentiate signal from background Includes chapter-ending summaries of Key Points to Remember, bringing beginners up to speed with any seasoned veteran in the field Thoughtfully tackles the molecular basis if IHC and ISH, along with application of that knowledge to improving the techniques is significant

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.

"This second edition volume provides detailed protocols that address the challenges of signal-transduction IHC. This book delves into chapters that discuss the nature of signal transduction phenomena and approaches to making phosphor-specific antibodies, as well as numerous bona fide methods methods on digital imaging techniques, preservation of tissue targets, multicolor detection, flow cytometry, lipophagy analysis, apoptosis, and the combination of IHC with in situ hybridization. Written in the highly successful Methods in Molecular Biologyseries format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Thorough and comprehensive, Signal Transduction Immunohistochemistry: Methods and Protocols, Second Edition is a valuable resource to both novices and experts in other fields of biomedical research who need advice on IHC protocols to study signal transduction. This book will also be useful for researchers in academia, government labs, and the biotech industry." -- OCLC.

Lorette Javois' timely new 2nd edition revises and updates her widely acclaimed collection of step-by-step immunocytochemical methods, one that is now used in many biological and biomedical research programs. The methods are designed for researchers and clinicians who wish to visualize molecules in plant or animal embryos, tissue sections, cells, or organelles. In addition to cutting-edge protocols for purifying and preparing antibodies, light microscopic analysis, confocal microscopy, FACS, and electron microscopy, this revised edition contains many new methods for applying immunocytochemical techniques in the clinical laboratory and in combination with in situ hybridization.