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Use of Ice Recrystallization Inhibition Assays to Screen for Compounds That Inhibit Ice Recrystallization

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Cryopreservation and Freeze-Drying Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2180))

Abstract

Ice recrystallization inhibition assays are used to screen for compounds that possess the ability to inhibit ice recrystallization. The most common of these assays are the splat cooling assay (SCA) and sucrose sandwich assay (SSA). These two assays possess similarities; however, they vary in their sample size, cooling rate, and the solution used to dissolve the analyte. In this chapter, both assay methods are described in detail, and we perform a direct comparison of the assays by evaluating the IRI activity of an antifreeze protein (AFP I). IRI activity is quantified by using ImageJ software to analyze ice crystals, and a quantitative value describing the efficiency of the inhibitor is generated. This analysis emphasizes the importance of choosing the right assay to measure IRI activity.

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Authors and Affiliations

  1. Department of Chemistry and Biomolecular Sciences, University of Ottawa, Ottawa, ON, Canada

    Anna A. Ampaw, August Sibthorpe & Robert N. Ben

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  1. Anna A. Ampaw
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  2. August Sibthorpe
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  3. Robert N. Ben
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Correspondence to Robert N. Ben .

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Editors and Affiliations

  1. Unit for Reproductive Medicine—Clinic for Horses, Biostabilization Laboratory—Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany, University of Veterinary Medicine Hannover, Hannover, Germany

    Willem F. Wolkers

  2. Unit for Reproductive Medicine—Clinic for Horses, University of Veterinary Medicine Hannover, Hannover, Germany

    Harriëtte Oldenhof

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Ampaw, A.A., Sibthorpe, A., Ben, R.N. (2021). Use of Ice Recrystallization Inhibition Assays to Screen for Compounds That Inhibit Ice Recrystallization. In: Wolkers, W.F., Oldenhof, H. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology, vol 2180. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0783-1_9

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  • DOI: https://doi.org/10.1007/978-1-0716-0783-1_9

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  • Publisher Name: Humana, New York, NY

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