Multimodal imaging has the potential for non-invasive assessment of imaging biomarkers that guide and monitor treatment of glioblastoma tumours. In this work, an imaging protocol for characterization of implanted patient-derived and murine glioblastoma tumours (GBM2, GBM14, and U87) was established using T2-weighted MRI, DWI, hyperpolarized 13C-pyruvate-lactate CSI, and 18F-FDG PET. Tumours were visible in T2-weighted MRI as high-signal regions with poorly defined borders. Compared with shams and non-tumour brain, all tumour lines had elevated ADC, 18F-FDG Ki, and lactate-to-pyruvate ratio. GBM2 had particularly high and variable lactate-to-pyruvate ratio, despite relatively low variability in ADC.