One of the major obstacles to the successful treatment of diffuse gliomas is their highly infiltrative property. It is one of the important therapeutic strategies to inhibit infiltration of glioma cells and make it possible to locally control a lesion. To achieve this, we first need to observe and describe detailed movement profiles of glioma cells in the brain tissue at the cellular level. Then we further need to determine extra and intracellular molecules that play a key role in glioma cell invasion. Live cell imaging is a powerful technique to investigate the basic movement pattern of glioma cells as well as the effects of therapeutic interventions on their migration. In this review, we describe a technical aspect of live cell imaging with special regard to time-lapse video imaging and discuss the relevance of the methods to glioma studies.