Virginia opossums are most active at night. By day they rest in nests of leaves and grass, hidden away in crevices, hollow trees and sometimes in burrows. By night, the marsupials hunt for food. They are good climbers, using their prehensile tails to cling to branches.
Virginia opossums do not hibernate, but they do put on fat as the days shorten with the approach of autumn. They rely on this fat to keep them going during the periods of harshest winter weather, when they cannot get out to feed. In the very coldest parts of their range, these marsupials sometimes suffer frostbite on their naked tails and thin ears.
Mating takes place in both late winter and spring. The young are only 1cm long and underdeveloped when born. Over 20 are born, but the mother can only suckle 13 at once, so the weaker babies die.