Osteosarcoma is the most common primary malignant tumor of bone,particularly among children and adolescents.Advances in imaging,surgical techniques,and implants have dramatically reduced the need for amputation in the past three decades.Recently,in vivo fluorescence imaging in the second near-infrared window (NIR-Ⅱ,1,000-1,700 nm) shows impressive advantages of deeper tissue penetration and higher spatial resolution,which makes it a promising tool for the early diagnosis and post-operative observation of Osteosarcoma.To the best of our knowledge,this paper is the first time to develop a novel NIR-Ⅱ fluorescence probe conjugated with an osteosarcoma targeted oligopeptide for molecular tumor imaging in a xenograft orthotopic osteosarcoma mouse model.