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Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes

Ex vivo Imaging of T Cells in Murine Lymph Node Slices with Widefield and Confocal Microscopes

DOI:
10.3791/3054-v

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09:45 min

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July 15, 2011

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, , , , ,
1Institut Cochin,Université Paris Descartes, CNRS (UMR 8104), 2Inserm,U1016, Paris, France

Chapters

  • 00:05Title
  • 01:18Preparing Slices from Lymph Nodes
  • 03:58Isolation and Labeling of T cells from Mouse Lymph Nodes
  • 05:08Plating Labeled T cells onto Lymph Node Slices
  • 05:45Preparation of Microscope and Lymph Node Slices for Imaging
  • 06:27Imaging T cells with a Widefield Inverted Microscope
  • 07:24Imaging T cells with a Confocal Upright Microscope
  • 07:53Representative Images and Movies of T cell Accumulation, Recruitment, and Motility within Lymph Node Slices
  • 09:17Conclusion

Summary

Automatic Translation

Automatic Translation

This protocol describes a method to image fluorescent T cells introduced into lymph node slices. The technique permits real-time analyses of T cell migration with traditional widefield fluorescence or confocal microscopes.

Tags

Ex Vivo ImagingT CellsLymph Node SlicesWidefield MicroscopeConfocal MicroscopeMigration Of T CellsChemokinesTwo-photon MicroscopyQuantitative InformationInteractions With Other CellsSlice AssayFluorescently Labeled T CellsAcutely Prepared Lymph Node SliceReal-time ImagingNatural EnvironmentMechanisms Underlying T Cell Migration

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