Agarose Embedding and Vibratome Sectioning of Thymic Lobes
5:30
Preparation of Thymocytes for Overlaying
6:59
Overlaying Thymocytes onto the Thymic Slices
7:48
Results: Positive and Negative Selection of Thymocytes
9:29
Conclusion
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The overall goal of this procedure is to prepare thymic slices that in combination with flow cytometry, can be used to model positive and negative selection of developing T cells. This method can help answer key questions about T cell development,
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We describe the preparation of thymic slices that, in combination with flow cytometry, can be used to model positive and negative selection of developing T cells. Thymic slices can also be adapted for the in situ analysis of thymocyte migration, localization, and signaling via immunofluorescence and two-photon microscopy.