We used non-invasive ultrasonography to investigate seasonal changes in liver size of an obligate hibernator; the edible dormouse (Glis glis). We repeatedly measured liver size in dormice at two study sites, (1) an enclosure-housed colony and (2) a free-ranging population. We observed a significant increase in transverse liver size throughout the active season; however, this effect was most pronounced in animals with low body mass early in the active season. Ultrasonography imaging revealed that the content of fat in the liver (distributed in an unexpected pattern of discrete focal areas) visibly increased at the end of the active season both in the field and enclosure. Thus, the observed increase in liver size of dormice seems mainly related to fat accumulation. We found no principal differences in the time courses of transversal liver size between the two study sites. Our results support the view that non-invasive ultrasonography is an accurate method to study internal organ sizes, such as liver, in the laboratory and in the field.