Real-time monitoring of quorum sensing in 3D-printed bacterial aggregates using scanning electrochemical microscopy
Real-time monitoring of quorum sensing in 3D-printed bacterial aggregates using scanning electrochemical microscopy
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DOI：
10.1073/pnas.1421211111
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发表时间：
2014-12-23
影响因子：
11.1
通讯作者：
Whiteley, Marvin
中科院分区：
文献类型：
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作者：
Connell, Jodi L.;Kim, Jiyeon;Whiteley, Marvin
微生物在自然界中通常以小的、密集的聚集体形式存在，含有类似于10（1）-10（5）个细胞。这些聚集体不仅表现出不同的表型，包括对抗生素的抗性，而且还作为更大的生物膜群落的构建块。聚集在这些较大的社区显示非随机的空间组织，最近的证据表明，这种空间组织是健身的关键。研究单个聚集体以及空间组织聚集体仍然具有挑战性，因为与操纵小种群相关的技术困难。微3D打印是一种光刻技术，能够通过在单个细胞或小群体周围打印基于蛋白质的壁来原位产生聚集体。这种3D打印策略可以将细菌组织成复杂的排列，以研究空间和环境参数如何影响社会行为。在这里，我们结合了微型3D打印和扫描电化学显微镜（SECM）来探测细菌铜绿假单胞菌中的群体感应（QS）介导的通信。我们的研究结果表明，QS依赖的行为内观察到小至500个细胞的聚集体，然而，聚集体大于2,000细菌需要刺激QS在邻近的聚集体定位8 μ m远。这些研究提供了一个强大的系统来分析空间组织和聚集体大小对微生物行为的影响。
Microbes frequently live in nature as small, densely packed aggregates containing similar to 10(1)-10(5) cells. These aggregates not only display distinct phenotypes, including resistance to antibiotics, but also, serve as building blocks for larger biofilm communities. Aggregates within these larger communities display nonrandom spatial organization, and recent evidence indicates that this spatial organization is critical for fitness. Studying single aggregates as well as spatially organized aggregates remains challenging because of the technical difficulties associated with manipulating small populations. Micro-3D printing is a lithographic technique capable of creating aggregates in situ by printing protein-based walls around individual cells or small populations. This 3D-printing strategy can organize bacteria in complex arrangements to investigate how spatial and environmental parameters influence social behaviors. Here, we combined micro-3D printing and scanning electrochemical microscopy (SECM) to probe quorum sensing (QS)-mediated communication in the bacterium Pseudomonas aeruginosa. Our results reveal that QS-dependent behaviors are observed within aggregates as small as 500 cells; however, aggregates larger than 2,000 bacteria are required to stimulate QS in neighboring aggregates positioned 8 mu m away. These studies provide a powerful system to analyze the impact of spatial organization and aggregate size on microbial behaviors.