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| Fluorescence microscopy studies of ice surface--antifreeze protein interactions |
| Topic:
9A Imaging and Optical Microscopy ; 1D Protein-Ligand Interactions |
Natalya Pertaya, PhD1, Carlos L. Di Prinzio, PhD1, Erik Thomson2, Larry Wilen, PhD1, John Wettlaufer, PhD2, Peter L. Davies, PhD3, Ido Braslavsky, PhD1.
1Ohio University, Athens, OH, USA, 2Yale University, New Haven, CT, USA, 3Queen's University, Kingston, ON, Canada. |
| Presentation Number: 409-Pos |
| Poster Board Number: B248 |
| Certain species of fish and other animals have evolved specialized antifreeze proteins (AFP) that prevent their blood from freezing. These naturally-occurring proteins and synthetic proteins modeled after them have many potential applications in agriculture, food preservation, cryobiology and biomedical science. To realize these applications, a deep understanding of the mechanisms of antifreeze/ice-surface interactions is required, motivating intense study in this field for over 30 years. Despite a large amount of progress, many important issues remain unresolved. Here we describe a new experimental approach to study the interaction between AFPs and ice using fluorescence microscopy combined with a unique ice growth/flow cell. After conjugating green fluorescent protein (GFP) to type III AFP, we imaged the fluorescence signal around ice crystals that emerged from the cooled AFP-GFP solution. We observed an enhanced fluorescence signal at the edge of the crystal indicating a higher concentration of the protein at the ice surface. We also designed a temperature-controlled flow cell with a well-defined temperature gradient and observed a dramatic change in the growth morphology of polycrystalline ice when the ice AFP was introduced into an initially pure system. The influence of antifreeze protein on the grain boundaries will be used to investigate the change in surface energy in the presence of these proteins. Further developments of these methods will permit the direct imaging of the location and concentration of the AFPs on ice surfaces under extremely well-defined conditions of growth, temperature, solute flow. The system of the AFPs and ice can be used as a model platform to understand bio-mineralization processes and thus is important in future nanotechnology applications. Supported by CIHR, the Bosack and Kruger Foundation, Yale and Ohio Universities. |
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