Development of an Automated Frozen Sample Aliquotter

描述(由申请人提供)： 生物信息库是癌症转化研究的宝贵资源，在学术和工业环境中都有许多这样的信息库。虽然这些储存库很有价值，但它们也不能幸免于成本限制，存储生物标本(例如血清和血浆)的方法涉及到在使用更多的小瓶存储样本之间的基本成本权衡，每个小瓶的体积(L到L)适合于检测或以更大的体积(2毫升或4毫升)存储样本，以节省冷冻箱空间。第一种方法避免了下游等分，只有一个冷冻/解冻周期，但需要人工对新鲜样品进行等分，而且体积效率低。第二种方法在需要样品时需要等分，因此样品在进行研究时经历了冻结/解冻循环，但在体积上是有效的。还采用一种混合方法，其中新鲜样品最初以体积效率格式存储，直到它们被请求进行研究，然后以体积效率低的格式返回存储，以避免随后的冻结/解冻循环。该项目将开发一种自动化仪器，它将提供一种新的方法，将体积效率高的存储与单次冻结/解冻周期相结合。这台仪器将从冷冻样品中提取等分，而不解冻样品。这种方法不仅可以减少操作这些储存库的成本，同时消除了与混合方法相关的第二个冷冻/解冻周期，而且因为它是自动化的，所以它将增加处理样本的吞吐量，将1000个样本研究的6半周减少到1周，这在护士健康研究中是典型的。该团队已经证明了以下能力：(1)在加工过程中将样品保持在-70摄氏度；(2)从-80摄氏度冷冻的生理盐水中提取等分；(3)在加工过程中控制结霜。开发R21项目和里程碑是为了解决与该仪器相关的关键技术风险，R33项目将提供适合护士健康研究使用的原型仪器，以便为他们的合作者处理样本。此时的设计将被很好地记录下来，使其能够在我们的实验室小批量生产，用于其他生物仓库。

DESCRIPTION (provided by applicant): Biorepositories are a valuable resource in translational research for cancer and there are many such repositories in both academic and industrial settings. While these repositories are valuable, they are not immune from cost constraints and the approach to storing biological specimens (e.g., serum and plasma) involves a fundamental cost tradeoff between storing the samples in a larger number of vials each with volumes (100 ¿l to 400 ¿l) suitable for assaying or storing the samples in larger volumes (2 ml or 4 ml) to save freezer space. The first approach avoids downstream aliquotting and has only one freeze/thaw cycle but requires labor to aliquot the fresh sample and is volumetrically inefficient. The second approach requires aliquotting when the samples are requested and as a result the sample experiences a freeze/thaw cycle when it is processed for a study but is volumetrically efficient. A hybrid approach is also pursued where fresh samples are initially stored in a volumetrically efficient format until they are requested for a study and then returned to storage in the volumetrically inefficient format to avoid subsequent freeze/thaw cycles. This project will develop an automated instrument that will offer a new approach, combining the volumetrically efficient storage with the single freeze/thaw cycle. This instrument will extract aliquots from frozen samples without thawing the samples. Not only will this approach reduce the cost of operating these repositories while eliminating the second freeze/thaw cycle associated with the hybrid approach, but because it is automated it will increase the throughput in processing samples, reducing the 6 1/2 weeks to one week for a 1000 sample study, which is typical for the Nurses' Health Study. This team has demonstrated the ability to: (1) maintain the sample at -70 oC during processing; (2) extract aliquots from normal saline frozen at -80 oC, and (3) manage frost buildup during the procedure. The R21 project and the milestones have been developed to address the key technical risks associated with this instrument and the R33 project will deliver a prototype instrument that is suitable for use by the Nurses' Health Study to process samples for their collaborators. The design at this point will be documented well enough to enable its production in our lab in small quantities for use in other biorepositories.