STAR Protocols (Dec 2024)

Protocol to quantify the activation dynamics of tumor-associated T cells in mice by functional intravital microscopy

  • Shannon N. Geels,
  • Claire Murat,
  • Alexander Moshensky,
  • Shivashankar Othy,
  • Francesco Marangoni

Journal volume & issue
Vol. 5, no. 4
p. 103310

Abstract

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Summary: Tumor-associated T cells orchestrate cancer rejection after checkpoint blockade immunotherapy. T cell function depends on dynamic antigen recognition through the T cell receptor (TCR) resulting in T cell activation. Here, we present an approach to quantify the dynamics and magnitude of tumor-associated T cell activation at multiple time points in living mice using the genetically encoded calcium reporter Salsa6f and functional intravital microscopy (F-IVM). Our protocol allows researchers to measure the activation dynamics of various immune cells in vivo.For complete details on the use and execution of this protocol, please refer to Geels et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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