Ecology and life characteristics of overwintering larvae of the European corn borer (Ostrinia nubilalis Hbn.) (Lep.: Crambidea) are partly unexplored due to their hidden lifestyle. In plant protection research the best way to study these phenomena is to apply less used, non-destructive, in vivo methods. The objective of our CT survey was to examine the factors influencing the location of the overwintering O. nubilalis larvae in maize stalks. The findings obtained by CT-analysis can be used for monitoring the presence and location of O. nubilalis larvae in the stalk, as well as both their displacement and movement. Our results showed that both the location and the distance from the brace root of O. nubilalis larvae were significantly influenced by the sampling time, the number of larvae per plant, the stalk diameter and finally the prevailing temperature. The location of the larvae situated nearest to the brace roots (first larvae) was significantly lower in stalks containing several larvae, than those where only a single larva was found in the stalk. The thickness of stalks was related to the simultaneous presence of more larvae, and to the ground level position of the first larvae. These overwintering larvae were located closer to the brace root (and to the soil), possibly because of having moved downwards inside the stalk, where the temperature is slightly milder than in the upper part of the stalk.