Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species.
Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species.
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水生物种玻璃化环原型的三维打印。
DOI：
10.1089/zeb.2017.1520
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发表时间：
2019
期刊：
影响因子：
2
通讯作者：
Tiersch,TerrenceR
中科院分区：
文献类型：
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作者：
Tiersch,NolanJ;Childress,WilliamM;Tiersch,TerrenceR
玻璃化是一种快速冷冻样品的低温保存方法，同时形成无定形固体（“玻璃”），通常体积较小（μL）。该项目的目标是通过三维（3D）打印创建基于椭圆形回路的开放式玻璃化冷冻设备，可以有效地使用和存储。玻璃化的努力可以受益于3D打印的应用，为了开始整合这项技术，我们解决了四个主要变量：热塑性长丝类型、环长度、环高度和装载方法。我们的目标是：（1）设计具有不同尺寸的玻璃化环;（2）打印原型环用于测试;（3）评估装置的加载方法;以及（4）对多种装置配置的玻璃化响应进行分类。各种配置是使用3D CAD（计算机辅助设计）软件进行数字设计的，原型设备是使用MakerBot®3D打印机生产的。用于生产器械的热塑性长丝是丙烯腈丁二烯苯乙烯（ABS）和聚乳酸（PLA）。玻璃化装置的特征在于用不同的加载方法（移液或浸没）形成的膜体积。冷冻膜进行分类，以确定玻璃化质量：零（不透明，或丰富的结晶冰形成）;一（半透明，或部分玻璃化），或两个（透明，或大量玻璃化，玻璃）。使用公开的玻璃化溶液进行实验。通过移液加载比通过浸没更可靠地形成冷冻膜，但浸没产生更少的填充问题并且更快速。产生最高玻璃化水平的环路设计能够快速传热，并且通常被表征为更长并且由更少的层（高度）组成。3D打印可以帮助玻璃化方法和研究的标准化，还可以在需要时提供快速设计和制造定制设备的能力。
Vitrification is a method of cryopreservation that freezes samples rapidly, while forming an amorphous solid (“glass”), typically in small (μL) volumes. The goal of this project was to create, by three-dimensional (3D) printing, open vitrification devices based on an elliptical loop that could be efficiently used and stored. Vitrification efforts can benefit from the application of 3D printing, and to begin integration of this technology, we addressed four main variables: thermoplastic filament type, loop length, loop height, and method of loading. Our objectives were to: (1) design vitrification loops with varied dimensions; (2) print prototype loops for testing; (3) evaluate loading methods for the devices; and (4) classify vitrification responses to multiple device configurations. The various configurations were designed digitally using 3D CAD (Computer Aided Design) software, and prototype devices were produced with MakerBot®3D printers. The thermoplastic filaments used to produce devices were acrylonitrile butadiene styrene (ABS) and polylactic acid (PLA). Vitrification devices were characterized by the film volumes formed with different methods of loading (pipetting or submersion). Frozen films were classified to determine vitrification quality: zero (opaque, or abundant crystalline ice formation); one (translucent, or partial vitrification), or two (transparent, or substantial vitrification, glass). A published vitrification solution was used to conduct experiments. Loading by pipetting formed frozen films more reliably than by submersion, but submersion yielded fewer filling problems and was more rapid. The loop designs that yielded the highest levels of vitrification enabled rapid transfer of heat, and most often were characterized as being longer and consisting of fewer layers (height). 3D printing can assist standardization of vitrification methods and research, yet can also provide the ability to quickly design and fabricate custom devices when needed.