Effects of 12 biotic and abiotic factors on the freezing point of the sub-Antarctic springtail, Tullbergia antarctica, were investigated. Repeated cooling of individual springtails five times resulted in very similar freezing points suggesting that ice nucleation in this freeze-susceptible species is likely to be initiated by intrinsic factors rather than being a stochastic event. Mean supercooling point (SCP) was influenced by cooling protocol, showing a linear increase in mean SCP with cooling rates from 8 to 0.1 °C min−1. However, the opposite effect (decreasing SCP) was seen with slower cooling. Slower rates may be ecologically realistic and allow time for appropriate physiological and biochemical changes. Feeding and food presence in the gut had no effect on SCP, and there was no correlation between the ice nucleating activity of bacteria isolated from the guts and the whole springtail SCP. Habitat altitude and diurnal light and temperature regimes also had no effect on SCP. There was no correlation between the cryoprotectant concentration of fresh animals and their SCP, but experimental desiccation resulted in increased osmolality and decreased SCP, although with considerable individual variation. The most significant influence on SCP was associated with ecdysis. As springtails cease feeding for a period either side of ecdysis, shedding the entire gut lining, moulting may be an efficient mechanism of clearing the gut of all ice nucleating material. This previously unrecognised relationship between ecdysis, cold tolerance and seasonal survival tactics may play an important role in over-winter survival of some arthropods.