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Proc Natl Acad Sci U S A. 2012 May 8;109(19):7181-6. doi: 10.1073/pnas.1200894109. Epub 2012 Apr 25.

Monitoring single-cell bioenergetics via the coarsening of emulsion droplets.

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  • 1Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloïdes et Matériaux Divisés, Université Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France. laurent.boitard@espci.fr

Abstract

Microorganisms are widely used to generate valuable products, and their efficiency is a major industrial focus. Bioreactors are typically composed of billions of cells, and available measurements only reflect the overall performance of the population. However, cells do not equally contribute, and process optimization would therefore benefit from monitoring this intrapopulation diversity. Such monitoring has so far remained difficult because of the inability to probe concentration changes at the single-cell level. Here, we unlock this limitation by taking advantage of the osmotically driven water flux between a droplet containing a living cell toward surrounding empty droplets, within a concentrated inverse emulsion. With proper formulation, excreted products are far more soluble within the continuous hydrophobic phase compared to initial nutrients (carbohydrates and salts). Fast diffusion of products induces an osmotic mismatch, which further relaxes due to slower diffusion of water through hydrophobic interfaces. By measuring droplet volume variations, we can deduce the metabolic activity down to isolated single cells. As a proof of concept, we present the first direct measurement of the maintenance energy of individual yeast cells. This method does not require any added probes and can in principle apply to any osmotically sensitive bioactivity, opening new routes for screening, and sorting large libraries of microorganisms and biomolecules.

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