Effective management of tree-hollow-dependent wildlife requires a sound knowledge of the characteristics of the trees used for shelter or breeding. We used radio-tracking to identify the den trees of squirrel gliders (Petaurus norfolcensis) in south-east Queensland (Qld) and north-east New South Wales (NSW). Squirrel gliders used dead trees as well as 13 species of living tree for dens across the two locations. Dead trees accounted for a large percentage of dens (54% of 48 dens in Qld, and 50% of 18 dens in NSW) despite comprising only 3–10% of the forest (trees >20 cm diameter at breast height (dbh)) at each location. This preference is largely due to dead trees being more likely to contain hollows, accounting for 26–44% of available hollow-bearing trees. Mean den tree size (dbh) was 48.9 ± 2.4 cm in Qld and 62.8 ± 5.6 cm in NSW. Den entrance height averaged 6.8 ± 1.2 m in Qld and 11.9 ± 1.3 m in NSW. Fissures in the trunk and holes in branches were the most common of six hollow types used. At one location branch end hollows were ignored relative to their availability. Den entrances varied in size (2.5–12 cm wide) but most were ≤5 cm in diameter. Entrance size of hollows appears to be the hollow attribute of most importance to squirrel gliders. Monitoring of these den trees over several years revealed the collapse of three dead den trees at each location, which is equivalent to an annual loss of 3% of den trees. Further research is needed to determine whether this will lead to a future shortage of den trees.