Abstract: Orthotopic xenograft model of human brain cancer cells is a good preclinical model for evaluation of antitumor compounds. In the present study, an orthotopic xenograft model of U87MG-mCherry-luc was established in Balb/c nude mice and the tumor growth was monitored via imaging technology including magnetic resonance imaging (MRI), in vivo imaging (IVI) and micro-CT. Furthermore, the model was evaluated with a positive drug temozolomide. Our data suggest that integrated imaging technology including MRI, IVI and micro-CT in orthotopic xenograft model can be used to facilitate monitoring of cancer progression and evaluate antitumor activity of drugs against glioma.