A digital microfluidic-microcoil NMR discovery platform to elucidate, monitor and understand cumulative environmental stress

这项提议旨在开发一种基于核磁共振(核磁共振)的新技术，可以实时检测水生细胞和生物体内的生化应激。核磁共振可以说是现代研究中最强大的工具，它提供了关于体内结构以及分子间和分子内相互作用的前所未有的分子信息。然而，核磁共振的致命弱点是其相对较低的灵敏度。该提案与Bruker Biospin合作，将减少核磁共振检测电路(小至20微米)，为细胞、鸡蛋和小型生物等质量有限的样品提供数量级的灵敏度提升。为了解决与这种小样品微线圈相关的样品处理和培养障碍，核磁共振将直接与数字微流体集成在一起。数字微流控是一个开放的电极阵列，夹在两个平板之间，允许混合、萃取、滴定、分级、层析、培养在芯片上自动进行，并允许对单独可寻址的质量有限的样品进行多个样品操作。**这项研究将用于检测、解释和确定复杂环境混合物中的关键应激源，并阐明应激源之间的累积效应。监测恢复的研究可以确定有机体在暴露后是否恢复到动态平衡，从而区分暂时性的“休克”反应和永久性的代谢变化，后者是确定环境政策的关键门槛。DMF-核磁共振为天然样品(废水、湖水等)的在线分馏提供了可能。与实时毒性测试相结合，以确定复杂天然混合物中最有问题的亚成分。这项技术应该能够确定：环境是否对生命安全？；确定造成问题的毒素类别；通过自动混合/暴露概况发现新的累积亚致命影响，并代表着监测总体环境变化的关键工具。这种早期预警系统对于在大范围的环境影响和生态系统服务中断之前确定应激源至关重要。

This proposal aims to develop a novel Nuclear Magnetic Resonance (NMR) based technology that can detect real-time biochemical stress in aquatic cells and organisms in-vivo. NMR is arguably the most powerful tool in modern research, providing unprecedented levels of molecular information on structures and inter- and intra-molecular interactions in-vivo. The Achilles heel of NMR however, is its relatively low sensitivity. In collaboration with Bruker Biospin the proposal will reduce NMR detection circuitry (as small as 20 µm) providing orders of magnitude increase in sensitivity for mass-limited samples such as cells, eggs, and small organisms. To solve the sample handling and culturing obstacles associated with such small sample micro-coil NMR will be directly integrated with digital microfluidics. Digital microfluidic is an open array of electrodes, sandwiched between two plates permitting mixing, extractions, titrations, fractionation, chromatography, culturing to be automated on-chip and permitting multiple sample manipulations on individually addressable mass-limited samples. **The research will be used to detect, explain and identify the key stressors in complex environmental mixtures and elucidate cumulative effects between stressors. Studies monitoring recovery can determine whether an organism returns to homeostasis after exposure thus differentiating between a temporary "shock" response from a permanent metabolic change, the later a key threshold for determining environmental policy. DMF-NMR provides the possibility for online fractionation of natural samples (waste water, lake water etc.) combined with real-time toxicity testing to identify the most problematic sub-components in complex natural mixtures. The technology should be able to determine if: An environmental is safe for life ?; identify the toxin category causing the problem; discover new cumulative sub-lethal effects via automated mixing/exposure profiling and represents a key tool to monitor environmental change in general. Such early warning systems are essential to identify stressors prior to widespread environmental impacts and ecosystem service disruption.