Department of Mechanical Engineering, University of California at Berkeley, Berkeley, CA, USA.
Department of Mechanical Engineering, University of California at Berkeley, Berkeley, CA, USA.
Cryobiology. 2022 Jun;106:91-101. doi: 10.1016/j.cryobiol.2022.03.003. Epub 2022 Mar 23.
Stable aqueous supercooling has shown significant potential as a technique for human tissue preservation, food cold storage, conservation biology, and beyond, but its stochastic nature has made its translation outside the laboratory difficult. In this work, we present an isochoric nucleation detection (INDe) platform for automated, high-throughput characterization of aqueous supercooling at >1 mL volumes, which enables statistically-powerful determination of the temperatures and time periods for which supercooling in a given aqueous system will remain stable. We employ the INDe to investigate the effects of thermodynamic, surface, and chemical parameters on aqueous supercooling, and demonstrate that various simple system modifications can significantly enhance supercooling stability, including isochoric (constant-volume) confinement, hydrophobic container walls, and the addition of even mild concentrations of solute. Finally, in order to enable informed design of stable supercooled biopreservation protocols, we apply a statistical model to estimate stable supercooling durations as a function of temperature and solution chemistry, producing proof-of-concept supercooling stability maps for four common cryoprotective solutes.
稳定的水相过冷具有作为人类组织保存、食品冷藏、保护生物学等领域的技术的巨大潜力，但由于其随机性，使其难以在实验室外进行转化。在这项工作中，我们提出了一种等容成核检测（INDe）平台，用于对 >1 mL 体积的水相过冷进行自动化、高通量的特性分析，从而能够统计有效地确定给定水体系中超冷稳定的温度和时间段。我们利用 INDe 研究了热力学、表面和化学参数对水相过冷的影响，并证明了各种简单的系统修改可以显著提高过冷稳定性，包括等容（恒体积）限制、疏水性容器壁以及甚至添加低浓度的溶质。最后，为了能够明智地设计稳定的过冷生物保存方案，我们应用统计模型来估计稳定过冷持续时间作为温度和溶液化学的函数，为四种常见的冷冻保护剂产生了概念验证的过冷稳定性图。
