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Immunology and Infection

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JoVE Journal Immunology and Infection
In Vitro Canine Neutrophil Extracellular Trap Formation
 

In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy

Article DOI: 10.3791/58083-v • 09:45 min • August 24th, 2018
August 24th, 2018 •
1, 2
1Department of Veterinary Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, 2Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis

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We describe methods to isolate canine neutrophils from whole blood and visualize NET formation in live neutrophils using fluorescence microscopy. Also described are protocols to quantify NET formation and citrullinated histone H3 (citH3) expression using immunofluorescence microscopy.

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In Vitro Canine Neutrophil Extracellular Trap Formation Dynamic Analysis Quantitative Analysis Fluorescence Microscopy NET Formation NET Components Mild Peroxidase Citrullinated Histone H3 Blood Collection Jugular Vein Sodium Heparin Tubes Anticoagulant Mixing Clots Red Cell Aggregates Dulbecco's PBS 3%Dextran Aggregation Sedimentation Erythrocytes Leukocyte-rich Plasma Density Gradient Centrifugation
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