Immunocytochemical Methods and Protocols / Edition 2by Lorette C. Javois
Pub. Date: 06/13/2008
Publisher: Springer-Verlag New York, LLC
Lorette Javois' timely new 2nd edition of Immunocytochemistry Methods 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 ContentsAntibody Preparation: Overview of Antibody Use in Immunocytochemistry. Purification of Antibodies Using Ammonium Sulfate Fractionation or Gel Filtration. Purification of Antibodies Using Affinity Chromatography. Purification of Antibodies Using Protein A-Sepharose and FPLC. Conjugation of Fluorochromes to Antibodies. Biotinylation of Antibodies. Tissue Preparation for Light Microscopic Analyses. Overview of Cell Fixation and Permeabilization. Preparation of Frozen Sections for Analysis. Processing of Cell-Touch Preparations. Processing of Cytological Specimens. Processing of Tissue-Culture Cells. Processing of Tissue Specimens. Light Microscopic Detection Systems. Overview of Fluorophores. Direct Immunofluorescent Labeling of Cells. Fluorescence Labeling of Surface Antigens on Attached or Suspended Tissue Culture Cells. Fluorescence Labeling of Intracellular Antigens in Attached or Suspended Cells. Fluorescent Labeling of Surface of Intracellular Antigens in Whole Mounts. Overview of Antigen Detection Through Enzymatic Activity. The Peroxidase-Antiperoxidase (PAP) Method. The Avidin-Biotin Complex (ABC) Method. Labeled Avidin Binding (LAB) Method. Avidin-Biotin Labeling of Cellular Antigens in Cryostat Sectioned Tissue. ELISA on Attachment-Dependent or Suspension Grown Cells. Use of Immunogold with Silver Enhancement. Fluorescence Activated Cell Sorter (FACS) Analyses. Overview of Flow Cytometry and Fluorescent Probes for Cytometry. Tissue Disaggregation. Indirect Immunofluorescent Labeling of Viable Cells. Indirect Immunofluorescent Labeling of Fixed Cells. Fluorescent Labeling of DNA. Deparaffinization and Processing of Pathological Material. Assay for Phagocytosis. Assay forFilamentous Actin. Assay for Chemoattractant Binding. Assay for Oxidative Metabolism. Colloidal Gold Detection Systems for Electron Microscopic Analyses. Fixation and Embedding. Preparation of Colloidal Gold. Conjugation of Colloidal Gold to Proteins. Colloidal Gold/Streptavidin Methods. Pre-Embedding Labeling Methods. Postembedding Labeling Methods. Photography. Overview of Fluorescence Photography. Special Applications. Overview of Fluorescence Analysis with the Confocal Microscope. Overview of Laser Microbeam Applications as Related to Antibody Targeting. Nucleic Acid Immunocytochemistry: In Situ Hybridization to Human Chromosomes of an Alkaline Phosphatase- Labeled Centromeric Probe. Fluorescence In Situ Hybridization Using Whole Chromosome Library Probes. Overview of Automated Immunostainers. Overview of Immunohistochemical Approaches to the Differential Diagnosis of Tumors. Index.
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