Antibody Preparation -- Overview of Antibodies for Immunochemistry -- to the Purification of Antibodies -- Antibody Purification: Ammonium Sulfate Fractionation or Gel Filtration -- Antibody Purification: Ion-Exchange Chromatography -- Antibody Purification: Affinity Chromatography - Protein A and Protein G Sepharose -- Conjugation of Fluorochromes to Antibodies -- Biotinylation of Antibodies -- Tissue Preparation for Light Microscopic Analysis -- Cell Fixatives for Immunostaining -- Permeabilization of Cell Membranes -- Preparation of Frozen Sections for Analysis -- Processing of Cytological Specimens -- Processing of Tissue Culture Cells -- Processing of Tissue Specimens -- Heat-Induced Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections -- Bright Field Detection Systems -- Fluorochromes: Properties and Characteristics -- Direct Immunofluorescent Labeling of Cells -- Fluorescence Labeling of Surface Antigens of Attached or Suspended Tissue-Culture Cells -- Fluorescence Labeling of Intracellular Antigens of Attached or Suspended Tissue-Culture Cells -- Fluorescent Visualization of Macromolecules in Drosophila Whole Mounts -- Overview of Conventional Fluorescence Photomicrography -- Overview of Confocal Microscopy -- Overview of Laser Microbeam Applications as Related to Antibody Targeting -- Immuno-Laser Capture Microdissection of Rat Brain Neurons for Real Time Quantitative PCR -- Overview of Antigen Detection Through Enzymatic Activity -- The Peroxidase-Antiperoxidase (PAP) Method and Other All-Immunologic Detection Methods -- The Avidin-Biotin Complex (ABC) Method and Other Avidin-Biotin Binding Methods -- Avidin-Biotin Labeling of Cellular Antigens in Cryostat-Sectioned Tissue -- Multiple Antigen Immunostaining Procedures -- Immunoenzymatic Quantitative Analysis of Antigens Expressed on the Cell Surface (Cell-ELISA) -- Use of Immunogold with Silver Enhancement -- Fluorescence-Activated Cell Sorter (FACS) Analyses -- Overview of Flow Cytometry and Fluorescent Probes for Flow Cytometry -- Tissue Disaggregation -- Indirect Immunofluorescent Labeling of Viable Cells -- Indirect Immunofluorescent Labeling of Fixed Cells -- Fluorescent Labeling of DNA -- Deparaffinization and Processing of Pathologic Material -- Colloidal Gold Detection Systems for Electron Microscopic Analysis -- Fixation and Embedding -- Preparation of Colloidal Gold -- Conjugation of Colloidal Gold to Proteins -- Colloidal Gold/Streptavidin Methods -- Pre-embedding Labeling Methods -- Postembedding Labeling Methods -- The Clinical Laboratory -- The Clinical Immunohistochemistry Laboratory: Regulations and Troubleshooting Guidelines.

The landscape of immunocytochemistry has significantly evolved in past years. Once a laborious technique with limited application, the introduction of new detection systems and instrumentation has facilitated the widespread use of immunocytochemical methods. In the post-genomic era, the use of immunocytochemical methods has grown to include the identification of a broad spectrum of antigens including proteins, carbohydrates and lipids from virtually every organism. In Immunocytochemical Methods and Protocols, Third Edition, expert researchers explore the modern methods employed in the field today, now widely used to identify, in situ, various components of cells and tissues in both normal and pathological conditions, and to demonstrate the localization of subcellular components. Chapters provide methods that are designed to be general in their application and to appeal to investigators across all disciplines, including those in research and clinical settings. Composed in the highly successful Methods in Molecular Biology™ series format, chapters contain a brief introduction, step-by-step methods, a list of necessary materials, and a Notes section which shares tips on troubleshooting and avoiding known pitfalls. Comprehensive and ground-breaking, Immunocytochemical Methods and Protocols, Third Edition, brings together a variety of methods that are normally limited to applications in a specific area, providing a unique and vital guide for researchers and clinicians alike.