Flow-through Luminescence Analyzer for Real-time Evaluation (FLARE)

FLARE is a real-time, in-line, flow-through fluorescence spectrometer that operates using four excitation wavelengths ranging from the DUV (~250nm) through the visible to excite fluorescence in microbial proteins, metabolites (e.g., NAD(P)H, siderophores), and pigments. The use of multiple excitation wavelengths allows for the detection of many different molecular species (including background contaminants) which would otherwise not all be accessible and/or as distinguishable with a single excitation wavelength.FLARE draws heritage from another flow-through fluorescence instrument developed by LMT which uses a single excitation wavelength (265nm) and six emission bands to detect microbial protein fluorescence. This predecessor has been deployed and tested aboard the VALKYRIE cryobot at the Matanuska Glacier (AK) where it monitored, in real time, microbial cells in the generated melt water at concentrations below 50 cells/mL with good precision. By expanding the number of excitation wavelengths and significantly increasing the number of detection channels to 16 or even 32, FLARE will be able to excite a much larger number of compounds and be better equipped to spectroscopically distinguish between them. This will allow FLARE to detect cells (e.g., via proteins) and, by measuring the fluorescence from tell-tale fluorophores such as NAD(P)H (which only fluoresces in a reduced state) and siderophores (e.g., pyoverdin), inform as to the metabolic activity or possible pathogenicity of the microbes.Since FLARE is a continuously-monitoring flow-through system, it can detect larger eukaryotic cells such as protists and fungi at very low levels (lt;lt; 1 cell/mL). Their relatively large biomass will cause very large distinguishable spikes in the data. Based on performance of its predecessor, we expect that FLARE will be able to meet the monitoring requirements of NASA#39;s microbiological limits of detection for potable water sources.