Development of Germplasm Resources for Preservation of Aquatic Models

斑马鱼(Danio Rerio)是世界各地生物医学研究和实验室的强大模型 世界上已经产生了数以万计的突变和转基因品系。维护这些 作为活鱼的有价值的基因类型是昂贵的，有风险的，超出了种群中心的能力。AS 这样，冷冻保存已经成为一种必要，这些遗传资源中的大多数现在都是 保存为质量不一致的样品，用最基本的技术冷冻。质量 控制没有以任何系统的方式进行，重复性差，协议 不规范。在受精过程中出现问题和失败，导致品系丢失是很常见的 这需要重新创建，导致设施浪费大量的时间和精力。这在很大程度上 由于忽视质量控制节省了时间和金钱的错误观念。然而，更确切地说 这些成本通过浪费的存储空间向下游转移 减少受精。这种普遍缺乏质量控制的情况使大量的 对生物医学研究的投资风险很大。因此，我们的长期目标是提供 廉价、普遍和系统的质量控制导致开发 基于社区的超低温保存标准。这将使来自 从单个实验室到大型综合存储库，为遗传基因提供保护 在生物医学研究中至关重要的资源。为了实现这一目标，我们建议提高可靠性和 通过以下方式提供对可重复性和标准化的常规访问，从而提高效率 微细加工(“芯片实验室”)和三维(3-D)打印设备的发展 包括从样本采集到受精的所有过程步骤。具体目标 是：1)开发和测试可供研究实验室使用的微型制造设备，网址为 以低成本和低成本改进精子密度和活动率的评估和研究 考虑到这些因素对可再生性和总体成功的影响 超低温保存。2)开发和测试3D打印设备，可用于改进 冷冻过程的重复性。这些设备将解决传统的 超低温保存和玻璃化冷冻，适用于单个样品和混合批次。3) 对这些设备进行生物测试，以改进设计和功能，并改进 质量评估的重现性，以使研究实验室能够支持或提交 它们被转移到库存中心和种质资源库。这将提供一个基于社区的 通过系统地纳入设备来保护遗传资源的方法， 适用于各种活动范围和规模的准则和标准。

Zebrafish (Danio rerio) is a powerful model in biomedical research and laboratories around the world have produced tens of thousands of mutant and transgenic lines. Maintaining these valuable genotypes as live fish is expensive, risky, and beyond the capacity of stock centers. As such, cryopreservation has become a necessity and most of these genetic resources are now maintained as samples of inconsistent quality frozen with rudimentary techniques. Quality control has not been practiced in any systematic way, reproducibility is poor, and protocols are not standardized. It is common to have problems and failures in fertilization resulting in lost lines that need to be recreated, causing facilities to waste considerable time and effort. This is largely due to the false notion that neglecting quality control saves time and money. However, rather than being reduced, these costs are shifted downstream through wasted storage space and reduced fertilization. This pervasive lack of quality control has placed the substantial investments in biomedical research at great risk. Therefore, our long-term goal is to provide inexpensive, universally available and systematic quality control leading to development of community-based standards for cryopreservation. This will enable reliable contributions from individual laboratories to large comprehensive repositories providing protection for genetic resources pivotal in biomedical research. To achieve this, we propose to improve reliability and efficiency by providing routine access to reproducibility and standardization through continued development of microfabricated (“laboratory on a chip”) and 3-dimensional (3-D) printed devices encompassing all process steps from sample collection through fertilization. The Specific Aims are to: 1) Develop and test microfabricated devices that can be used by research laboratories at low effort and cost to improve assessment and study of sperm concentration and motility with respect to the effects of these factors on reproducibility and the overall success of cryopreservation. 2) Develop and test 3-D printed devices that can be used to improve the reproducibility of the freezing process. These devices will address conventional cryopreservation and vitrification, and be useful for single samples and pooled batches. 3) Perform biological testing of these devices to refine design and function and improve the reproducibility of quality assessment to enable research laboratories to back up lines or submit them to stock centers and germplasm repositories. This will provide a community-based approach for protection of genetic resources through systematic incorporation of devices, guidelines and standards applicable across a full range of activity scopes and scales.