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

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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.