Molecular events promoting tumourigenesis and anticancer therapeutic strategies have been intensively studied in tumour cell culture models. In the past few years, non-invasive bioluminescence imaging (BLI) has emerged as a powerful strategy for the validation of cell culture findings in animal models of cancer. BLI allows for repetitive and exceptionally sensitive real-time monitoring of a disease course, as well as of tumour response to therapeutic interventions in an individual animal. This review discusses the application of BLI to cancer research in general and to the area of experimental neuro-oncology in particular.