The overwintering temperatures of ants might well be elevated due to climate change. We studied whether the overwintering temperature affects the survival of the queens and whole colonies of the black garden ant, Lasius niger (Linnaeus, 1758). In two consecutive years (2009, 2010) we collected mated, colony founding queens (n = 280) from the urban area of Turku, Finland. Half of the queens overwintered in +7 to +8 °C and the other half in +2 °C. After the overwintering period, we determined their survival rate and measured the body fat content, body size and immune defence (encapsulation rate) of overwintering queens. Using the same setup, we studied the survival of 1-year-old L. niger colonies (queen & workers). Overwintering at a lower temperature (+2 °C) decreased the survival of workers. The survival of colony founding queens differed between years, but unlike with workers, the overwintering temperature did not affect their survival: neither in the colony experiment nor in the single queen experiment. All of the surviving queens managed to produce their worker offspring at the same rate. The relative amount of body fat of queens was higher for those who overwintered at a lower temperature, which is likely a result of lower energy consumption. We did not detect differences in the encapsulation rate between the temperature treatment groups. The ability of colony founding queens to tolerate wide overwintering temperature variations present in urban environments may explain the success of the colony in urban areas. As the colony grows, the overwintering chambers may extend more deeply into the ground. Thus, workers may not have to cope with such cold conditions as colony founding queens.