GIRAULT, Mathias; BENEYTON, Thomas; PEKIN, Deniz; BUISSON, Lionel; BICHON, Sabrina; CHARBONNIER, Celine; DEL AMO, Yolanda; BARET, Jean-Christophe

doi:10.1021/acs.analchem.8b00234

Centre de Recherche Paul Pascal [CRPP]

BENEYTON, Thomas

Centre de Recherche Paul Pascal [CRPP]

PEKIN, Deniz
Centre de Recherche Paul Pascal [CRPP]

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Article de revue

Ce document a été publié dans

Analytical Chemistry. 2018, vol. 90, n° 6, p. 4174 - 4181

Résumé en anglais

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 water-in-oil droplets and measure alkaline phosphatase activity at the single-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. The advantages of subnanoliter-scale compartmentalization in droplet-based microfluidics provide a precise characterization of a population with single-cell resolution. Our results highlight the key role of cell physiology to efficiently access dissolved organic phosphorus.< Réduire

Mots clés en anglais

Microfluic

Vesicles

Droplet

URI

https://oskar-bordeaux.fr/handle/20.500.12278/199353

DOI

http://dx.doi.org/10.1021/acs.analchem.8b00234

Project ANR

Initiative d'excellence de l'Université de Bordeaux - ANR-10-IDEX-0003

Unités de recherche

EPOC : Environnements et Paléoenvironnements Océaniques et Continentaux - UMR 5805

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High-Content Screening of Plankton Alkaline Phosphatase Activity in Microfluidics