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In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy

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

8,516 Views

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09:45 min

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August 24, 2018

DOI:

10.3791/58083-v

DOI:
10.3791/58083-v

•

09:45 min
August 24, 2018

•

4 Views
,
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

Summary

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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 VitroCanineNeutrophilExtracellular TrapFormationFluorescence MicroscopyNETPeroxidaseCitrullinated Histone H3BloodJugular VeinAnticoagulantDextranDensity Gradient CentrifugationErythrocytesPolymorphonuclear CellsCell Culture BufferTrypan BluePoly-L-lysineFluorescence Microscopy

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Li, R. H., Tablin, F. In Vitro Canine Neutrophil Extracellular Trap Formation: Dynamic and Quantitative Analysis by Fluorescence Microscopy. J. Vis. Exp. (138), e58083, doi:10.3791/58083 (2018).

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