Ultra-rapid cooling under the appropriate conditions will produce vitrification, a glass-like state used to cryopreserve small sample volumes, but there are a number of major technical drawbacks impeding application of vitrification to germplasm of aquatic species. These include a lack of suitable devices, and poor reproducibility and comparability among studies due to a lack of standardization. We used 3-dimensional (3-D) printing to produce a viewing pedestal coupled with a classification system to rapidly assess frozen film quality of vitrification loops. Classification time declined with practice from 2.1 ± 0.3 sec to 1.5 ± 0.2 sec (after 200 assessments), and assessments were consistently made in < 2.5 sec. Classifications should be reported with representative images allowing harmonization for quality control. This approach permits rapid classification and can be applied for development of methods including evaluation of vitrification solution components, concentrations of solution and target cells, and configurations and volumes of new devices. Future studies should address the custom fabrication of 3-D printed vitrification devices for use with aquatic species and other applications.

中文翻译：

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水生物种薄膜玻璃化的标准化评估

在适当条件下的超快速冷却会产生玻璃化，这是一种用于冷冻小体积样品的玻璃状状态，但存在许多主要的技术缺陷，阻碍了玻璃化在水生物种种质中的应用。这些包括缺乏合适的设备，由于缺乏标准化，研究之间的可重复性和可比性较差。我们使用 3 维 (3-D) 打印来制作观察台，并结合分类系统快速评估玻璃化循环的冷冻膜质量。分类时间随着实践从 2.1 ± 0.3 秒下降到 1.5 ± 0.2 秒（200 次评估后），并且评估始终在 < 2.5 秒内进行。分类应与代表性图像一起报告，以便协调质量控制。这种方法允许快速分类，可用于开发方法，包括评估玻璃化溶液成分、溶液和目标细胞的浓度，以及新设备的配置和体积。未来的研究应该解决用于水生物种和其他应用的 3-D 打印玻璃化装置的定制制造。