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Live Imaging of Resident T-Cell Migration in Human Lymphoid Tissue Slices Using Confocal Microscopy

  • Emmanuel DonnadieuEmail author
  • Yvonne Michel
  • Martin-Leo Hansmann
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

Abstract

In order to mount a potent immune response, immune cells must move actively through tissues. As an example, T-cell need to migrate within lymph nodes in order to scan the surface of many dendritic cells and recognize rare expressed antigens. The recent development of improved imaging approaches, such as two-photon microscopy, and the use of powerful mouse models have shed light on some of the mechanisms that regulate the migration of immune cells in many organs. Whereas such systems have provided valuable insights, they do not always predict human responses. In human, our knowledge in the field mainly comes from a description of fixed tissue samples. However, these studies lack a temporal dimension since samples have been fixed. In order to overcome some of these limitations, we describe, in this methodology chapter, an experimental system of fresh human adenoid slices to monitor the dynamics of resident T-lymphocytes that have been stained with directly-coupled fluorescent antibodies. Combined with confocal fluorescent imaging, this preparation offers an effective approach to imaging immune cells in a three-dimensional (3D) human lymphoid tissue environment.

Key words

T-cells Motility Vibratome Immunostaining Confocal microscopy 

Notes

Acknowledgments

We wish to thank Nadine Flinner (Frankfurt Institute for Advanced Studies) for help and during the establishment of this technique.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Emmanuel Donnadieu
    • 1
    • 2
    • 3
    Email author
  • Yvonne Michel
    • 4
  • Martin-Leo Hansmann
    • 4
    • 5
  1. 1.Département Immunologie, Inflammation, et InfectionINSERM, U1016, Institut CochinParisFrance
  2. 2.CNRS, UMR8104ParisFrance
  3. 3.Université Paris Descartes, Sorbonne Paris CitéParisFrance
  4. 4.Dr. Senckenbergisches Institut für Pathologie, Klinikum der Johann Wolfgang Goethe-UniversitätFrankfurt am MainGermany
  5. 5.Frankfurt Institute for Advanced Studies (FIAS)Frankfurt am MainGermany

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