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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Monitoring single-cell bioenergetics via the coarsening of emulsion droplets
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Monitoring single-cell bioenergetics via the coarsening of emulsion droplets

机译：通过乳剂液滴的粗化监测单细胞生物能

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摘要

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.
机译：微生物被广泛用于生产有价值的产品，其效率是工业的主要重点。生物反应器通常由数十亿个细胞组成，可用的测量结果仅反映了种群的整体表现。但是，细胞的贡献并不相等，因此，通过监控这种种群内多样性可以优化过程。迄今为止，由于无法探测单细胞水平的浓度变化，因此这种监测仍然很困难。在这里，我们通过在浓缩的反相乳液中利用包含活细胞的小滴与周围空滴之间的渗透驱动水通量来解除这种限制。通过适当的配方，与初始营养素（碳水化合物和盐）相比，排泄的产品在连续疏水相中的溶解度要高得多。产品的快速扩散会引起渗透失配，由于水通过疏水界面的扩散较慢，渗透失配会进一步放松。通过测量液滴的体积变化，我们可以推断出新陈代谢的活动可以归结为分离的单个细胞。作为概念的证明，我们提出了单个酵母细胞维持能量的首次直接测量。该方法不需要添加任何探针，并且原则上可以应用于任何对渗透敏感的生物活性，开辟了新的筛选途径，并可以对大型的微生物和生物分子文库进行分类。

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  • 作者

    L Boitard; D. Cottinet; C. Kleinschmitt; N. Bremond; J. Baudry; G. Yvert; J. Bibette;

  • 作者单位

    Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloides et Materiaux Divises, Universite Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France;

    Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloides et Materiaux Divises, Universite Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France;

    Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloides et Materiaux Divises, Universite Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France;

    Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloides et Materiaux Divises, Universite Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France;

    Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloides et Materiaux Divises, Universite Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France;

    Ecole Normale Superieur Lyon, Laboratoire de Biologie Moleculaire de la Cellule, Universite Lyon 1, Unite Mixte de Recherche 5239, 46 Allee d'ltalie, 69007 Lyon, France;

    Ecole Superieure de Physique et de Chimie Industrielles, Paris Tech, Laboratoire de Colloides et Materiaux Divises, Universite Pierre et Marie Curie, Paris 06, Unite Mixte de Recherche 7195, 10 Rue Vauquelin, 75231 Paris, France;

  • 收录信息 美国《科学引文索引》(SCI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

    biosensors; metabolism;

    机译：生物传感器代谢;

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