Extracellular macromolecules associated with Antarctic sea ice diatoms were previously shown to have ice-binding activities. The function of these ice-active substances (IASs) has not been identified. Here we show that two of the IASs have a strong ability to inhibit the recrystallization of ice, possibly signifying a cryoprotectant function. To test this possibility, two species of marine diatom (one Antarctic and one temperate) were subjected to a single freeze-thaw cycle (approximately 20h at -4 or -5 degrees C) in the presence or absence of IAS. Viability, based on a double staining technique, was 15-29% higher in the presence of IAS. Etching of single crystal ice hemispheres grown from dilute IAS solutions indicated that the IASs bind to specific faces of ice and are incorporated into the ice lattice. Together, these results suggest that the IASs acts as a cryoprotectant, probably through some ice-binding mechanism.