In Vivo Tumor Hypoxia Imaging

A cocktail of either AF546-pHLIP + IR800-pHLIP (5 nmol each) or that of IR800-pHLIP + Cy5.5-RaftRGD (5 and 10 nmol, respectively) was administered to the mice and the mice were examined at 4 h post-injection. Pimonidazole, a commonly used exogenous hypoxia marker in histochemistry (1.5 mg, Hypoxyprobe-1 Omni Kit; Hypoxyprobe, Burlington, MA, USA), was administered intravenously to the mice 1 h before euthanizing. Tumors were removed and cryo-sectioned in sequential sections (10 µm thickness) as previously described [40 (link)]. Sections were stored in the dark at −80 °C until subsequent experiments.