Enhanced Control over Ice Nucleation Stochasticity Using a Carbohydrate Polymer Cryoprotectant

Bruno M. Guerreiro, Anthony N. Consiglio, Boris Rubinsky, Matthew J. Powell-Palm, Filomena Freitas

Research output: Contribution to journalArticlepeer-review

6 Scopus citations


Metastable supercooling has emerged as a transformative technique for ice-free biopreservation, but issues of stability inherent to the stochastic nature of ice formation have thus far limited its translation out of the laboratory. In this work, we explore the influence of the bio-based carbohydrate polymer FucoPol on aqueous supercooling using an isochoric nucleation detection technique. We show that FucoPol, a high-molecular-weight, fucose-rich polysaccharide, which has previously been shown to reduce average ice crystal sizes after nucleation, also induces a concentration-dependent stabilization of metastable supercooled water, as evidenced by both a significant reduction in nucleation stochasticity (i.e., the spread in temperatures over which the system will nucleate upon cooling) and a corresponding increase in the predicted induction time of nucleation. FucoPol is found to confine the stochasticity of ice nucleation to a narrow, well-defined band of temperatures roughly one-third as wide as that of pure water under identical conditions. Importantly, this substantial reduction in stochasticity is accompanied by only a minimal (<1 °C) change in the average nucleation temperature, suggesting that this effect is distinct from colligative freezing point depression. Reducing and characterizing the stochasticity of aqueous supercooling is essential to the engineering design of practical biopreservation protocols, and the results reported herein suggest that high-viscosity polymer systems may provide a powerful and largely unexplored lever by which to manipulate metastable-equilibrium phase change kinetics at subzero temperatures.

Original languageEnglish (US)
JournalACS Biomaterials Science and Engineering
Issue number5
StateAccepted/In press - 2022

Bibliographical note

Funding Information:
This work received financial support from the National Science Foundation (NSF) Graduate Research Fellowship under Grant No. DGE 1752814, the NSF Engineering Research Center for Advanced Technologies for Preservation of Biological Systems (ATP-Bio) under NSF EEC Grant No. 1941543, and national funds from FCT─Fundação para a Ciência e a Tecnologia, I.P. (Portugal), in the scope of projects UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences─UCIBIO, LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy─i4HB, UID/QUI/50006/2013 of LAQV-REQUIMTE and UID/CTM/50025 of CENIMAT/I3N. B.M. Guerreiro also acknowledges PhD grant funding by Fundação para a Ciência e a Tecnologia, FCT I.P. (SFRH/BD/144258/2019) and supporting personal funding from Fulbright Portugal (21-066), Fundação Luso-Americana para o Desenvolvimento, FLAD (Proj. 2021/0070─G-2021-0052), and The Company of Biologists (JCSTF2105556).

Publisher Copyright:
© 2022 American Chemical Society.


  • biopolymer
  • cryobiology
  • FucoPol
  • nucleation stochasticity
  • supercooling
  • Carbohydrates
  • Temperature
  • Cryoprotective Agents/chemistry
  • Water/chemistry
  • Polymers

PubMed: MeSH publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Journal Article


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