Immunocytochemical methods and protocols

Part I. Antibody Preparation -- Overview of Antibody Use in Immunocytochemistry / Su-Yau Mao, Lorette C. Javois, and Ute M. Kent -- Purification of Antibodies Using Ammonium Sulfate Fractionation or Gel Filtration / Ute M. Kent -- Purification of Antibodies Using Ion-Exchange Chromatography / Ute M. Kent -- Purification of Antibodies Using Affinity Chromatography / Ute M. Kent -- Purification of Antibodies Using Protein A-Sepharose and FPLC / Ute M. Kent -- Conjugation of Fluorochromes to Antibodies / Su-Yau Mao -- Biotinylation of Antibodies / Su-Yau Mao -- Part II. Tissue Preparation for light Microscopic Analyses -- Overview of Cell Fixatives and Cell Membrane Permeants / Melissa A. Melan -- Preparation of Frozen Sections for Analysis / Gary L. Bratthauer -- Processing of Cytological Specimens / Gary L. Bratthauer -- Processing of Tissue Culture Cells / Gary L. Bratthauer -- Processing of Tissue Specimens / Gary L. Bratthauer -- Antigen Retrieval for Immunohistochemical Reactions in Routinely Processed Paraffin Sections / Laszlo Krenacs, Tibor Krenacs, and Mark Raffeld -- Part III. Light Microscopic Detection Systems -- Overview of Fluorochromes / J. Michael Mullins -- Direct Immunofluorescent Labeling of Cells / Lorette C. Javois -- Fluorescence Labeling of Surface Antigens on Attached or Suspended Tissue-Culture Cells / Mark C. Willingham -- Fluorescence Labeling of Intracellular Antigens in Attached or Suspended Tissue-Culture Cells / Mark C. Willingham -- Fluorescent Labeling of Surface or Intracellular Antigens in Whole-Mounts / Lorette C. Javois -- TUNEL Assay for Apoptotic Cells / Virginia M. Heatwole -- Overview of Fluorescence Analysis with the Confocal Microscope / Liana Harvath -- Overview of Laser Microbeam Applications as Related to Antibody Targeting / P. Scott Pine -- Overview of Fluorescence Photomicography / Lorette C. Javois and J. Michael Mullins -- Overview of Antigen Detection Through Enzymatic Activity / Gary L. Bratthauer -- The Peroxidase-Antiperoxidase (PAP) Method and Other All-Immunologic Detection Methods / Gary L. Bratthauer -- The Avidin Biotin Complex (ABC) Method and Other Avidin Biotin Binding Methods / Gary L. Bratthauer -- Avidin Biotin Labeling of Cellular Antigens in Cryostat Sectioned Tissue / Mark Raffeld and Elaine S. Jaffe -- Multiple Antigen Immunostaining Procedures / Laszlo Krenacs, Tibor Krenacs, and Mark Raffeld -- ELISA on Attachment Dependent or Suspension Grown Cells / Julie Ann Brent -- Use of Immunogold with Silver Enhancement / Constance Oliver -- Part IV. Fluorescence Activated Cell Sorter (FACS) Analyses -- Overview of Flow Cytometry and Fluorescent Probes for Cytometry / Robert E. Cunningham -- Tissue Disaggregation/ Robert E. Cunningham -- Indirect Immunofluorescent Labeling of Viable Cells / Robert E. Cunningham -- Indirect Immunofluorescent Labeling of Fixed Cells / Robert E. Cunningham -- Fluorescent Labeling of DNA / Robert E. Cunningham -- Deparaffinization and Processing of Pathologic Material / Robert E. Cunningham -- Assay for Phagocytosis / Liana Harvath and Douglas A. Terle -- Assay for Filamentous Actin / Liana Harvath -- Assay for Chemoattractant Binding / Liana Harvath, Robert R. Aksamit, and Robert E. Cunningham -- Assay for Oxidative Metabolism / Liana Harvath and Douglas A. Terle -- Part V. Colloidal Gold Detection Systems For Electron Microscopic Analyses -- Fixation and Embedding / Constance Oliver and Maria Celia Jamur -- Preparation of Colloidal Gold / Constance Oliver -- Conjugation of Colloidal Gold to Proteins / Constance Oliver -- Colloidal Gold/Streptavidin Methods / Constance Oliver -- Pre-Embedding Labeling Methods / Constance Oliver -- Postembedding Labeling Methods / Constance Oliver -- Part VI. Applications For Nucleic Acid Localization By In Situ Hybridization -- Overview of In Situ Hybridization / Roland M. Nardone -- In Situ Hybridization to Human Chromosomes of an Alkaline Phosphatase-Labeled Centromeric Probe / Roland M. Nardone -- Fluorescence In Situ Hybridization Using Whole Chromosome Library Probes / Roland M. Nardone -- In Situ RT-PCR and Hybridization Techniques / Vishakha Thaker -- Part VII. Special Applications For the Clinical Laboratory -- Overview of the Clinical Immunohistochemistry Laboratory : Regulations and Troubleshooting Guidelines / Patricia A. Fetsch and Andrea Abati -- Overview of Immunohistochemical Approaches to the Differential Diagnosis of Tumors / Dennis M. Frisman -- Overview of Automated Immunostainers / Gilbert E. Herman, Edna A. Elfont, and Alton D. Floyd.