1st Edition

Manual of Immunological Methods

By Canadian Networking Copyright 1998

    The Manual of Immunological Methods represents the collaboration of the Canadian Network of Toxicology Centers, a non-profit network of university-based scientists dedicated to research, training, risk assessment, and communication. This manual provides detailed immunological methods that can be utilized by researchers or practitioners who want to enhance the successful application of this science. It emphasizes the need for continuously improving the quality of experiments performed and maintaining consistency in the results obtained.
    The Manual of Immunological Methods presents validated assays that are accompanied by a series of tools that will guide readers in performing their own laboratory work in the spirit of the Good Laboratory Practices. It covers specific methodologies including sample collection, preparation of cells, assay conditions, and data evaluation. Discussion of species differences and comparisons of a wide variety of species is an important feature. Both graduate students and practicing researchers in the areas of immunology and immunotoxicology will appreciate the Manual of Immunological Methods as a guide to standard operating procedures in the laboratory.

    Identification, Anaesthesia, and Euthanasia
    Identification of Laboratory Rodents by Ear Notching
    Identification of Fish by Fin Clipping
    Anaesthesia of Fish and Rodents
    Euthanasia of Rodents
    Euthanasia of Fish
    Working Sheet: Weight of Laboratory Animals
    Collection of Peripheral Blood Samples
    Reagents
    Materials and Equipment
    Procedure
    Removal of Organs
    Reagents
    Materials and Equipment
    Procedure
    Preparation of Cell Suspensions
    Extrusion of Coelomocytes in the Earthworm
    Hemocyte Collection in the Mollusk
    Cell Suspension from Peritoneal Exudate
    Isolation of White Blood Cells from Peripheral Blood
    Cell Suspensions from Lymphoid Organs
    Assessment of Cell Viability
    Determination of Cell Viability and Concentration by Trypan Blue Dye Exclusion
    Determination of Cell Viability and Cell Concentration with Ethidium Bromide and Acridine Orange
    Flow Cytometric Assessment of Cell Viability
    Working Sheet: Cell Viability and Concentration by Trypan Blue Dye Exclusion or Ethidium Bromide/Acridine Orange
    Cell Cryopreservation
    Reagents
    Materials and Equipment
    Procedure
    Special Recommendations
    Analysis of Results
    Species-Related Changes in the Protocol
    Technique-Related Changes in the Protocol: Phagocytosis with Yeast
    Statistical Tests
    Working Sheet: Phagocytic Function
    Oxidative Burst Assay Using Flow Cytometry
    Reagents
    Materials and Equipment
    Procedure
    Special Recommendations
    Analysis of Results
    Species-Related Changes in the Protocol
    Statistical Tests
    Working Sheet: Acquisition List
    Cell Cytotoxicity
    Natural Killer (NK) Cell Activity
    Antibody-Dependent Cellular Cytotoxicity
    Lymphokine-Activated Killer (LAK)
    Working Sheet: Acquistion List
    Lymphoblastic Transformation
    Reagents
    Materials and Equipment
    Procedure
    Analysis of Result
    Special Recommendations
    Species-Related Changes in the Protocol
    Working Sheet: Mitogenic Assay
    Determination of Antibody-Producing Cells to a Specific Antigen
    Liquid Plaque Forming Cell Assay in Mouse Model
    Agar Plaque Forming Cell Assay in Rat Model
    Working Sheet: Plaque Forming Cells (PFC)
    Mixed Lymphocyte Reaction (MLR)
    Reagents
    Materials and Equipment
    Procedure
    Analysis of Results
    Special Recommendations
    Species-Related Changes in the Protocol
    Working Sheet: Mixed Lymphocyte Reaction
    Intracellular Levels of Calcium Assay Using Flow Cytometry
    Reagents
    Materials and Equipment
    Procedure
    Special Recommendations
    Analysis of Results
    Species-Related Changes in the Protocol
    Statistical Tests
    Working Sheet: Acquisition List
    Phenotyping of Blood Mononuclear Cells
    Reagents
    Materials and Equipment
    Procedure
    Special Recommendations
    Analysis of Results
    Species-Related Changes in the Protocol
    Immunophenotype of Rat Peripheral Blood Lymphocytes
    Working Sheet
    Evaluation of Intracellular Level of Thiols
    Reagents
    Materials and Equipment
    Procedure
    Special Recommendations
    Analysis of Results
    Species-Related Changes in the Protocol
    Working Sheet: Acquisition List
    Apoptosis
    Reagents
    Materials and Equipment
    Procedure
    Analysis of Results

    Biography

    Canadian Networking (University of Guelph)