Surgical oncology often requires the use of contrast-enhanced cross-sectional imaging preoperatively to characterize solitary tumours and identify sentinel lymph nodes. Intraoperative optical guidance can effectively aid tissue-sparing tumour excision and locate sentinel lymph nodes. Nanotrast-CF800 (CF800) is a novel dual-modality contrast agent, which co-encapsulates iohexol and indocyanine green (ICG) within a liposomal nanoparticle. It was developed for preoperative and intraoperative imaging of solitary tumours and sentinel lymph node mapping and its efficacy has been demonstrated in preclinical animal models (Zheng et al. 2015). The objective of this study is to evaluate the safety profile of CF800 following intravenous administration in healthy dogs. Six research dogs were randomized into two groups. Group 1 received a low dose (1 mL/kg) and group 2 received a high dose (5 mL/kg). Dogs were placed under general anesthesia and a continuous rate infusion of CF800 was administered based on group allocation. Physiologic parameters including heart rate, respiratory rate, direct arterial blood pressure, cardiac output, and temperature were measured at set time points. Plasma concentrations of iohexol, ICG, and histamine were measured at set time points. Dogs underwent whole body computed tomography scans pre-injection, 2-, and 7-days post-injection (p.i.). Contrast enhancement was measured in select organ systems and great vessels at each time point. There were no significant changes in physiologic parameters following IV infusion of CF800 in all dogs. Plasma iohexol and ICG concentrations peaked at 1 day p.i., while histamine concentrations peaked at 30 minutes p.i. Significant contrast enhancement was noted within the liver, heart, aorta, and caudal vena cava on day 2 p.i., which was significantly different compared to baseline. Prolonged contrast retention within the liver was identified. Intravenous administration of CF800 was safe to use in healthy dogs with no significant systemic adverse effects.