Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
Anal Chem. 2010 Feb 15;82(4):1288-91. doi: 10.1021/ac9022742.
A major roadblock to the vitrification of cells is the requirement of high concentrations of cryoprotectant (CPA) chemicals and the damage caused by prolonged exposure of cells to these high concentrations above the glass transition temperature. These effects are minimized with controlled CPA loading. Certain organic oils, such as soybean oil, are made of triacylglycerols and are capable of dissolving small amounts of water, a property which is enhanced significantly as temperature is increased. This phenomenon was exploited here to accomplish temperature-controlled concentration of glycerol in single water droplets dispersed in the organic phase. Emulsions of aqueous solutions of glycerol in soybean oil were made and subjected to a temperature increase of 10 degrees C from room temperature. Upon increasing temperature, water dissolved into the oil, rendering the 15-20 microm droplets concentrated an average of 3.6 times and 2.6 times for 1 and 2 M starting concentrations, respectively, with the oil-insoluble glycerol in 90-110 s. This phenomenon could be used to dynamically concentrate CPAs within cell-containing droplets which may then be vitrified before being exposed to high temperatures for fatally long times.
细胞玻璃化的主要障碍是需要高浓度的冷冻保护剂（CPA）化学物质，以及细胞长时间暴露在高于玻璃化转变温度的高浓度下造成的损伤。通过控制 CPA 加载可以最小化这些影响。某些有机油，如豆油，由三酰基甘油组成，能够溶解少量的水，随着温度的升高，这种性质会显著增强。在这里，我们利用这一现象来实现单个分散在有机相中的水液滴中甘油的温度控制浓度。制备了甘油在大豆油中的水溶液乳液，并从室温升高 10°C。随着温度的升高，水溶解到油中，使 15-20 微米的液滴分别平均浓缩 3.6 倍和 2.6 倍，起始浓度分别为 1 和 2 M，而 90-110 s 内 90-110 s 内油不溶性甘油。这种现象可用于在含有细胞的液滴内动态浓缩 CPAs，然后在暴露于高温下很长时间之前将其玻璃化。
