Intravital multiphoton microscopy of the metabolic co-enzymes NAD(P)H and FAD (optical metabolic imaging, or OMI) provides label-free imaging of metabolic changes in vivo. Since the metabolism of tumor and immune cells is associated with cancer progression, we aim to study metabolic changes during a triple-combination immunotherapy regimen that cures murine melanoma tumors. Our results demonstrate that intravital OMI can capture tumor and T cell autofluorescence intensity and lifetime changes during immunotherapy. Overall, this technology enables analysis of single cell metabolic changes in vivo to provide insight for immunotherapy development.
Current methods to assess immune cell function use labels that are limiting for time-course studies of immune cell behavior in tumors. Here, we use multiphoton imaging of NAD(P)H and FAD, co-enzymes of metabolism, in T cells and macrophages within the tumor microenvironment. T cells alter their metabolism in response to tumor-like pH, glucose, and lactic acid levels, while macrophages alter their metabolism during tumor-stimulated migration. These results indicate that multiphoton autofluorescence imaging is a powerful label-free method to monitor immune cell metabolism within single cells in the tumor microenvironment.
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