High-Content Screening of Plankton Alkaline Phosphatase Activity in Microfluidics

Abstract : One way for phytoplankton to survive orthophosphate depletion is to utilize dissolved organic phosphorus by expressing alkaline phosphatase. The actual methods to assay alkaline phosphate activity-either in bulk or as a presence/absence of enzyme activity-fail to provide information on individual living cells. In this context, we develop a new microfluidic method to compartmentalize cells in 0.5 nL waterin-oil droplets and measure alkaline phosphatase activity at thesingle-cell level. We use enzyme-labeled fluorescence (ELF), which is based on the hydrolysis of ELF-P substrate, to monitor in real time and at the single-cell level both qualitative and quantitative information on cell physiology (i.e., localization and number of active enzyme sites and alkaline phosphatase kinetics). We assay the alkaline phosphatase activity of Tetraselmis sp. as a function of the dissolved inorganic phosphorus concentration and show that the time scale of the kinetics spans 1 order of magnitude...................
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Submitted on : Tuesday, April 3, 2018 - 4:14:50 PM
Last modification on : Friday, July 12, 2019 - 1:52:29 PM

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Mathias Girault, Thomas Beneyton, Deniz Pekin, Lionel Buisson, Sabrina Bichon, et al.. High-Content Screening of Plankton Alkaline Phosphatase Activity in Microfluidics. Analytical Chemistry, American Chemical Society, 2018, 90 (6), pp.4174 - 4181. ⟨10.1021/acs.analchem.8b00234⟩. ⟨hal-01757331⟩

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