Zhang Yuzhen, Cai Yumin, Zeng Lingbin, Liu Peng, Ma Luyan Z, Liu Jintao
Center for Infectious Disease Research School of Medicine Tsinghua University Beijing 100084 China.
Tsinghua-Peking Center for Life Sciences Beijing 100084 China.
Small Sci. 2022 Sep 20;2(10):2200047. doi: 10.1002/smsc.202200047. eCollection 2022 Oct.
Bacterial biofilms play essential roles in ecological environments and in human health. The spatial heterogeneity of biofilms is crucial to their resistance and collective behavior, while quantitative analysis of these biofilm-specific features is limited. Here, a microfluidic approach is developed to address this issue. Through a special design of microfluidic chamber and spatially controllable bacteria seeding, biofilms are cultivated with customized semi-2D structure, which enables quantitative measurements of spatially heterogeneous features with time-lapse microscopy. The advantages of the proposed method are demonstrated via two examples on biofilm homeostasis and stress response, respectively, where the functionally important spatiotemporal dynamics is delineated. In homeostasis, it is found that biofilms use spatially organized extracellular matrix to preserve iron chelators within their boundaries while maximizing free sharing within the community. In stress response, the spatial distribution of antibiotics in biofilms and how a change in energy metabolism leads to redistribution of drugs over space are elucidated. The proposed method enables cultivating biofilms formed by a wide range of species and even multiple biofilms, which provides a tractable approach to understanding the spatiotemporal features of biofilms formed by environmentally and clinically important bacteria.
细菌生物膜在生态环境和人类健康中发挥着重要作用。生物膜的空间异质性对其抗性和集体行为至关重要，而对这些生物膜特异性特征的定量分析却很有限。在此，开发了一种微流控方法来解决这一问题。通过微流控腔室的特殊设计和空间可控的细菌接种，培养出具有定制半二维结构的生物膜，这使得能够通过延时显微镜对空间异质性特征进行定量测量。分别通过生物膜稳态和应激反应的两个例子证明了该方法的优势，其中描绘了功能上重要的时空动态。在稳态中，发现生物膜利用空间组织的细胞外基质在其边界内保留铁螯合剂，同时在群落内最大限度地实现自由共享。在应激反应中，阐明了抗生素在生物膜中的空间分布以及能量代谢的变化如何导致药物在空间上的重新分布。所提出的方法能够培养由多种物种甚至多种生物膜形成的生物膜，这为理解由环境和临床重要细菌形成的生物膜的时空特征提供了一种易于处理的方法。
