Manual of Immunological Methods  book cover
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1st Edition

Manual of Immunological Methods



ISBN 9780849385582
Published July 2, 1998 by CRC Press
160 Pages

 
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Book Description

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.

Table of Contents

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

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