Microbial Nanoculture as an Artificial Microniche.
Microbial Nanoculture as an Artificial Microniche.
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DOI：
10.1038/srep30578
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发表时间：
2016-08-01
影响因子：
4.6
通讯作者：
Lee D
中科院分区：
文献类型：
--
作者：
Niepa TH;Hou L;Jiang H;Goulian M;Koo H;Stebe KJ;Lee D
微生物在小菌落中自组织，同时在保护性生物膜基质内转变为固着形式。为了能够详细研究这些小菌落内的微生物动力学，需要新的无柄培养系统来隔离细胞并模拟其复杂的生长条件和相互作用。我们提出了一种新的纳升规模的固着培养系统，很容易通过微流体使能制造实施。成千上万的这些纳米培养物可以很容易地产生和成像使用常规或共聚焦显微镜。每个纳米培养物开始时是几纳升的悬浮细胞液滴，由聚二甲基硅氧烷（PDMS）膜包裹。PDMS外壳提供持久的机械支撑，使长期研究成为可能，并且对小分子具有选择性渗透性，包括抗生素、信号分子和功能性荧光探针。因此，当小菌落在纳米培养物内成熟时，可以使用选定的探针对其进行应激或询问，以表征细胞生理特性、抗生素敏感性和拮抗相互作用。我们通过研究广泛的微菌落动态（包括直接和间接的细菌-真菌相互作用）来展示这个平台。这种多功能的新工具在解决与耐药性、慢性感染、微生物组动力学和抗生素发现相关的生物学问题方面具有广泛的潜力。
Microbes self-organize in microcolonies while transitioning to a sessile form within a protective biofilm matrix. To enable the detailed study of microbial dynamics within these microcolonies, new sessile culture systems are needed that sequester cells and mimic their complex growth conditions and interactions. We present a new nanoliter-scale sessile culture system that is easily implemented via microfluidics-enabled fabrication. Hundreds of thousands of these nanocultures can be easily generated and imaged using conventional or confocal microscopy. Each nanoculture begins as a several nanoliter droplet of suspended cells, encapsulated by a polydimethylsiloxane (PDMS) membrane. The PDMS shell provides long-lasting mechanical support, enabling long term study, and is selectively permeable to small molecules including antibiotics, signaling molecules and functional fluorescent probes. Thus, as microcolonies mature within the nanocultures, they can be stressed or interrogated using selected probes to characterize cell physiological properties, antibiotic susceptibilities, and antagonistic interactions. We demonstrate this platform by investigating broad ranges of microcolony dynamics, including direct and indirect bacterial-fungal interactions. This versatile new tool has broad potential for addressing biological questions associated with drug resistance, chronic infections, microbiome dynamics, and antibiotic discovery.