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Detection of Trypanosoma brucei Variant Surface Glycoprotein Switching by Magnetic Activated Cell Sorting and Flow Cytometry
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Immunology and Infection
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JoVE Journal Immunology and Infection
Detection of Trypanosoma brucei Variant Surface Glycoprotein Switching by Magnetic Activated Cell Sorting and Flow Cytometry

Detection of Trypanosoma brucei Variant Surface Glycoprotein Switching by Magnetic Activated Cell Sorting and Flow Cytometry

DOI:
10.3791/54715-v

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09:45 min

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October 19, 2016

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, , ,
1Laboratory of Lymphocyte Biology,Rockefeller University, 2Department of Biology,Harvey Mudd College, 3Masters School

Chapters

  • 00:05Title
  • 01:05Sample Harvesting
  • 02:05Magnetic Labeling
  • 04:05Magnetic Separation
  • 05:45Staining and Identification of Contaminants and Switchers
  • 07:32Results: Induced Double-strand DNA Breaks Increase the Frequency of Switchers in a Population
  • 09:01Conclusion

Summary

Automatic Translation

Automatic Translation

African trypanosomes grown in vitro undergo antigenic variation at a low rate, such that populations are made up of parasites expressing a dominant variant surface glycoprotein (VSG) type and a small population of "switched" variants. This protocol describes a fast method for detecting and quantifying these populations.

Tags

Keywords: Trypanosoma BruceiVariant Surface GlycoproteinVSG SwitchingMagnetic Activated Cell SortingFlow CytometryAntigenic VariationCell IsolationCell EnrichmentLister 427 StrainAnti-VSG AntibodyCell SeparationCell CultureCell CentrifugationCell LabelingMagnetic Beads

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