U1016, Paris, France

3 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

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

Hélène Salmon^1,2, Ana Rivas-Caicedo^1,2, François Asperti-Boursin^1,2, Camille Lebugle^1,2, Pierre Bourdoncle^1,2, Emmanuel Donnadieu^1,2

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

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.

Immunology and Infection

"Phagosome Closure Assay" to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy (TIRFM)

Florence Marie-Anaïs¹, Julie Mazzolini¹, Pierre Bourdoncle¹, Florence Niedergang¹

¹Inserm U1016, Institut Cochin, CNRS UMR 8104, Université Paris Descartes, Sorbonne Paris Cité

We describe an experimental setup to visualize with unprecedented high resolution phagosome formation and closure in three dimensions in living macrophages, using total internal reflection fluorescence microscopy. It allows monitoring of the base of the phagocytic cup, the extending pseudopods, as well as the precise site of phagosome scission.

Cancer Research

Ex Vivo Imaging of Resident CD8 T Lymphocytes in Human Lung Tumor Slices Using Confocal Microscopy

Elisa Peranzoni^*1, Houcine Bougherara^*1, Sarah Barrin^*1, Audrey Mansuet-Lupo^2,3, Marco Alifano⁴, Diane Damotte^2,3, Emmanuel Donnadieu¹

¹Institut Cochin, Inserm U1016-CNRS UMR8104, Université Paris Descartes, ²Department of Pathology, Paris Centre University Hospitals, Université Paris Descartes, ³INSERM U1138, Cancer and Immune Escape, Cordeliers Research Center, University Pierre and Marie Curie, ⁴Department of Thoracic Surgery, Paris Centre University Hospitals, University Paris Descartes

This protocol describes a method to image resident tumor-infiltrating CD8 T cells labeled with fluorescently coupled antibodies within human lung tumor slices. This technique permits real-time analyses of CD8 T cell migration using confocal microscopy.