About the Book
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.
Table of Contents:
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.
Review :
This second edition provides detailed protocols for immunocytochemical localization of tissue constituents. The book is well balanced and suited for basic and clinical research scientists...The editor has successfully compiled this book for the intended readers. The editor and contributors are credible...The protocol style format is useful for a first experiment. The illustrations are attractive and informative. The book as a whole is nicely done...I find this a very useful and practical addition to all fields of biology and medicine. It is very user friendly and should be a great reference for those designing experiments and writing. - Doody's Health Sciences Book Review Journal ...this book would make a valuable addition to laboratories currently involved in research using immunocytochemical techniques and would be particularly useful for those wishing to introduce these techniques into their research programmes...Given the depth of coverage of theory behind various techniques and the detailed notes that accompany each set of protocols, the book would also make an ideal teaching aid. - Molecular Biotechnology this excellent book on Methods and Protocols can be recommended to both investigators starting out in the field of immunocytochemistry and to basic and clinical researchers already working in the field because it contains a plethora of methods and protocols for the demonstration of molecules in many different normal and pathological situations and also under different experimental conditions. - Journal of Chemical Neuroanatomy
