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In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues

In Situ MHC-tetramer Staining and Quantitative Analysis to Determine the Location, Abundance, and Phenotype of Antigen-specific CD8 T Cells in Tissues

DOI:
10.3791/56130-v

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08:37 min

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September 22, 2017

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, ,
1Department of Veterinary and Biomedical Sciences,University of Minnesota

Capítulos

  • 00:05Título
  • 00:57Fresh Tissue Sectioning
  • 02:25Primary, Secondary, and Tertiary Antibody (Ab) Incubation
  • 04:37Sample Mounting and Confocal Microscopic Imaging
  • 05:47Results: Representative MHC-tetramer Staining
  • 07:25Conclusion

Summary

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Here, we describe a method that combines in situ MHC-tetramer staining with immunohistochemistry to determine localization, phenotype, and quantity of antigen-specific T cells in tissues. This protocol is used to determine the spatial and phenotypic characteristics of antigen-specific CD8 T cells relative to other cell type and structures in tissues.

Tags

Keywords: In SituMHC-tetramer StainingAntigen-specific CD8 T CellsQuantitative AnalysisImmunofluorescence StainingMicroscopy

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