Bacterial growth and adaptation in microdroplet chemostats.
Bacterial growth and adaptation in microdroplet chemostats.
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DOI：
10.1002/anie.201301524
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发表时间：
2013-08-19
影响因子：
16.6
通讯作者：
Garstecki, Piotr
中科院分区：
文献类型：
--
作者：
Jakiela, Slawomir;Kaminski, Tomasz S.;Cybulski, Olgierd;Weibel, Douglas B.;Garstecki, Piotr
我们在此描述了用于操纵和监测细菌群体的连续生长的微流体技术。由大约10个输入和输出通道组成的系统控制100多个微滴恒化器，并能够随着时间的推移独立地操纵每个微恒化器中的化学因子。在此，我们表征了微滴恒化器中细菌种群的动态和对一系列稳定或变化的抗生素浓度的细胞反应。这种方法允许对微生物生态学、生理学、进化和对化学环境的适应进行平行的长期研究。Leo Szilard [1]介绍恒化器是微生物学领域的一个里程碑。恒化器通过不断补充恒定体积的流体以维持特定浓度的细胞和生长因子来促进细菌、酵母和藻类的连续培养。[1，2] Chemostats促进了广泛的研究，包括微生物生态学，[3]捕食者-猎物动力学，[4]和耐药性的进化。[5，6]大量试剂的消耗和传统恒化器的重大操作挑战限制了它们的使用。恒化器的单相微流体版本使孵育体积最小化，[7-10]但仍受到其复杂性的限制：输入/输出控制的数量与恒化器的数量之间的比例妨碍了大规模并行化。单相微流体系统易于形成生物膜，这使得它们要么是一次性使用的设备[9]，要么需要额外的步骤来最大限度地减少细胞粘附。[8]液滴微流体[11]提供了一种独特的解决方案来创建许多并行恒化器。这项技术在微生物学中最早的例子是由约书亚·莱德伯格在近60年前首先证明的。[12]在此期间，微流体领域解决了许多与使用这种方法研究微生物相关的技术挑战。将细胞和营养物分隔在液体的微滴中可以降低操作许多平行恒化器的复杂性和成本。最近，细菌已经在通道中的液滴中在短时间间隔内孵育[13-17]，然而，在一系列完全可寻址的微滴中持续细胞生长数百代是不可能的。在此，我们描述了一种自动化微滴系统，该系统超越了现有的挑战，使用户能够操纵液滴的化学成分，用于长期的细菌研究。微流控系统（图1）执行三个功能：1）形成含有细胞，试剂和可溶性生长因子的微滴; 2）循环微滴用于细胞孵育和监测; 3）分裂和融合微滴以控制化学因子随时间的浓度。在用液体样品装载储器之后，我们使用压力源和外部阀来调节液体样品的流动。
We describe herein microfluidic technology for manipulating and monitoring continuous growth of populations of bacteria. A system consisting of approximately ten input and output channels controls more than 100 microdroplet chemostats and enables the manipulation of chemical factors in each microchemostat independently over time. Herein, we characterize the dynamics of bacterial populations in microdroplet chemostats and cellular responses to a range of stable or changing antibiotic concentrations. This method allows for parallel, long-term studies of microbial ecology, physiology, evolution, and adaptation to chemical environments. The introduction of the chemostat by Leo Szilard [1] was a milestone in the field of microbiology. Chemostats facilitate the continuous culture of bacteria, yeast, and algae by continuously replenishing a constant volume of fluid to maintain specific concentrations of cells and growth factors.[1, 2] Chemostats have facilitated a wide-range of studies, including microbial ecology,[3] predator–prey dynamics,[4] and the evolution of drug resistance.[5, 6] The consumption of large quantities of reagents and the significant operational challenges of traditional chemostats limit their use. Single-phase, microfluidic versions of chemostats minimize incubation volumes,[7–10] and yet are limited by their complexity: the proportionality between the number of input/output controls and the number of chemostats hamper large scale parallelization. Single-phase microfluidic systems are prone to biofilm formation, which makes them either singleuse devices [9] or requiring additional steps to minimize cell adhesion.[8] Droplet microfluidics [11] offer a unique solution to creating many parallel chemostats. The earliest example of this technology in microbiology was first demonstrated by Joshua Lederberg nearly 60 years ago.[12] In the interim, the field of microfluidics solved many of the technical challenges associated with using this approach to study microbes. Compartmentalizing cells and nutrients in microdroplets of liquid can reduce the complexity and cost of operating many parallel chemostats. Recently, bacteria have been incubated in droplets in channels over short time intervals,[13–17] however sustained cell growth over hundreds of generations in a series of fully addressable microdroplets has not been possible. Herein, we describe an automated microdroplet system that transcends existing challenges and enables users to manipulate the chemical composition of droplets for longterm bacterial studies. The microfluidic system (Figure1) performs three functions: 1) formation of microdroplets containing cells, reagents, and soluble growth factors; 2) cycling microdroplets for cell incubation and monitoring; and 3) splitting and fusing microdroplets to control the concentration of chemical factors over time. After loading the reservoirs with liquid samples, we used a source of pressure and external valves to regulate the flow of
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影响因子：
30.8
作者：
Toprak, Erdal;Veres, Adrian;Michel, Jean-Baptiste;Chait, Remy;Hartl, Daniel L.;Kishony, Roy
通讯作者：
Kishony, Roy
影响因子：
6.1
作者：
Baraban, Larysa;Bertholle, Fabien;Bibette, Jerome
通讯作者：
Bibette, Jerome
DOI：
10.1073/pnas.1106752109
发表时间：
2012-05-15
影响因子：
11.1
作者：
Leung, Kaston;Zahn, Hans;Hansen, Carl L.
通讯作者：
Hansen, Carl L.
DOI：
10.1073/pnas.70.8.2229
发表时间：
1973-01-01
影响因子：
11.1
作者：
PONGS, O;BALD, R;ERDMANN, VA
通讯作者：
ERDMANN, VA
影响因子：
6.1
作者：
Churski, Krzysztof;Kaminski, Tomasz S.;Garstecki, Piotr
通讯作者：
Garstecki, Piotr