Intravital Microscopy of T-B Cell Interactions in Germinal Centers

Changming Shih1, Hai Qi2

  • 1Laboratory of Dynamic Immunobiology, Department of Basic Medical Sciences, Tsinghua-Peking Center for Life Sciences, Institute for Immunology, School of Medicine, Tsinghua University, Beijing, 100084, China.

Dynamic imaging analyses of antigen-specific T-B interactions in germinal centers have advanced our understanding of the molecular mechanisms underlying affinity maturation and provided a wealth of information about how follicular helper T cells function in vivo. Here we describe a routine method to visualize fluorescence protein-expressing, antigen-specific T and B cells in germinal centers. The protocol for incorporating functional reporters or genetic perturbation of the T cells by retroviral transduction is also briefly described, using the FRET-based calcium reporter as an example.

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