In this project, we aim to translate a fluorine-19 (19F) MRI technology to visualize the tumor associated macrophage (TAM) burden in patients with recurrent head and neck squamous cell carcinoma (HNSCC). HNSCC is the sixth most common cancer worldwide. TAMs often comprise a significant volume fraction of tumor mass, and a high TAM burden in the tumor microenvironment is associated with poor therapeutic prognosis. In HNSCC patients developing locoregionally recurrent or metastatic disease, anti-PD-1 antibody checkpoint inhibitor therapy is currently the standard of care, however, the response rates remain poor (~20%). TAMs have been associated with immunosuppression and T-cell exclusion, which are associated with a low response rate to checkpoint inhibitors. Thus, there is an urgent need for biomarker probes that can non-invasively monitor TAMs in clinical trials to help stratify patients and rationally optimize therapeutic strategies. Towards this goal, we propose translating a fast-clearing perfluorocarbon (PFC) nanoemulsion as a 19F MRI tracer to assay TAMs in vivo. Following intravenous injection, the size and morphology of nonbiologic PFC nanoemulsion droplets make them susceptible to endocytosis by phagocytic immune cells, particularly macrophages. The in situ labeled cells accumulate at inflammatory lesions, and the resulting 19F MRI hot-spots can be quantified, where the signal is linearly proportional to the macrophage burden. In situ PFC macrophage labeling has been used in a multitude of preclinical inflammation models, including in the context of TAMs. However, no injectable, clinical PFC nanoemulsion imaging agent product exists despite the proven specificity and safety of these compositions. Building on this work, the proposal has three Specific Aims: Aim 1. Perfluorocarbon NE formulation. This aim will identify a lead formulation and collaborate with a contract manufacturing organization to produce a novel nanoemulsion composition (“TAM-Sense”) at lite