IDBR: Controlling Oxygen in Standard Multiwell Plates with a Microfabricated Add-on

氧是影响许多不同生物现象的关键代谢变量。然而，目前探测这一变量的工具效率低下，而且自细胞培养技术问世以来一直没有改变。然而，随着氧气越来越多地牵涉到许多信号通路中，启用一个简单的工具来探索这一点将极大地促进这些研究，并加速科学发现。在该项目中，将开发和优化一个平台，以独立控制多井板每口井内的局部气体浓度。该平台将由聚二甲基硅氧烷(PDMS)插件组成，该插件将嵌套到标准多孔板中。该插件是一个带有气体渗透膜的无源网络，旨在将气体输送到多孔板中的贴壁细胞培养物中。根据微通道中氧的浓度，插入件将充当氧气的汇或源，并将被浸入井的细胞介质中。气体将通过在PDMS膜上的简单扩散将气体输送到细胞单层，并溶解到多孔板的培养介质中。PDMS膜和细胞单层之间的这种紧密间距允许快速扩散，在几分钟内在井中施加稳定的氧气梯度。该设备不仅将能够控制氧气梯度，而且将导致一整套设备，使研究人员能够精确控制多孔板中的微环境，以满足各种不同的需求，该平台具有促进新的和令人兴奋的实验的潜力，通过在任何标准细胞生物学实验室都可以访问的实验室模型中复制生命系统(体内)中出现的氧气梯度，这是目前技术无法实现的。这一结果将对许多领域产生巨大影响。为了培养人们对科学和工程的兴趣，少数族裔小学生将在伊利诺伊大学芝加哥分校进行互动实验室活动。高危小学生将成为目标，与芝加哥女童子军合作，提供科学和工程方面的积极经验。计划评估将包括形成性反馈，这将使我们能够在从活动中学习时调整我们的方法，以及对所述目标是否已达到的总结性评估。

Oxygen is a key metabolic variable that influences many different biological phenomena. However, current tools to probe this variable are inefficient and have not changed since the dawn of cell culture techniques. Yet, as oxygen is increasingly implicated in many signaling pathways, enabling a simple tool to explore this would greatly facilitate these investigations and accelerate scientific discovery. In this project, a platform to independently control local gas concentrations within each well of a multiwell plate will be developed and optimized. The platform will consist of a polydimethylsiloxane (PDMS) insert that will nest into a standard multiwell plate. The insert is a passive network with a gas permeable membrane aimed to deliver gas to adherent cell cultures in multiwell plates. The insert will act as a sink or source of oxygen depending on the concentration of the oxygen in the microchannels and will be immersed in the cell media of the well. The gas will be delivered to the cell monolayer through simple diffusion of oxygen across a PDMS membrane and dissolve into the culture media of the multiwell plate. This close spacing between the PDMS membrane and cell monolayer allows for rapid diffusion to impose steady state gradients of oxygen across the well within minutes. The device will not only enable control of oxygen gradients, but will lead to a whole class of devices that will allow researchers to precisely control the microenvironment in multiwell plates to address a variety of The platform has potential to facilitate new and exciting experiments not possible with current techniques, by replicating the oxygen gradients that occur in living systems (in vivo) in a laboratory model (in vitro) accessible to any standard cell biology lab. The result would have a huge impact across many fields.To develop interest in science and engineering, underrepresented minority elementary students will be exposed to interactive laboratory activities at the University of Illinois at Chicago. At risk elementary school students will be targeted by working with the Girl Scouts of Chicago to provide positive experiences with science and engineering. Program evaluation will include both formative feedback, which will allow us to adjust our methods as we learn from the activities, and a summative appraisal of whether the stated goals have been met.