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dMSCC: a microfluidic platform for microbial single-cell cultivation of Corynebacterium glutamicum under dynamic environmental medium conditions
Lab on a Chip ( IF 6.1 ) Pub Date : 2020-10-23 , DOI: 10.1039/d0lc00711k
Sarah Täuber 1 , Corinna Golze , Phuong Ho , Eric von Lieres , Alexander Grünberger
Affiliation  

In nature and in technical systems, microbial cells are often exposed to rapidly fluctuating environmental conditions. These conditions can vary in quality, e.g., the existence of a starvation zone, and quantity, e.g., the average residence time in this zone. For strain development and process design, cellular response to such fluctuations needs to be systematically analysed. However, the existing methods for physically imitating rapidly changing environmental conditions are limited in spatio-temporal resolution. Hence, we present a novel microfluidic system for cultivation of single cells and small cell clusters under dynamic environmental conditions (dynamic microfluidic single-cell cultivation (dMSCC)). This system enables the control of nutrient availability and composition between two media with second to minute resolution. We validate our technology using the industrially relevant model organism Corynebacterium glutamicum. The organism was exposed to different oscillation frequencies between nutrient excess (feasts) and scarcity (famine). The resulting changes in cellular physiology, such as the colony growth rate and cell morphology, were analysed and revealed significant differences in the growth rate and cell length between the different conditions. dMSCC also allows the application of defined but randomly changing nutrient conditions, which is important for reproducing more complex conditions from natural habitats and large-scale bioreactors. The presented system lays the foundation for the cultivation of cells under complex changing environmental conditions.

中文翻译:

dMSCC:在动态环境培养基条件下用于谷氨酸棒杆菌微生物单细胞培养的微流控平台

在自然界和技术系统中,微生物细胞经常会暴露于快速变化的环境条件中。这些条件的质量(例如,饥饿区的存在)和数量(例如,,即该区域的平均停留时间。对于菌株开发和过程设计,需要系统地分析细胞对这种波动的反应。但是,现有的物理模拟快速变化的环境条件的方法在时空分辨率上受到限制。因此,我们提出了一种在动态环境条件下培养单细胞和小细胞簇的新型微流体系统(动态微流体单细胞培养(dMSCC))。该系统能够以秒至分钟的分辨率控制两种培养基之间的养分利用率和组成。我们使用行业相关模型生物谷氨酸棒杆菌验证我们的技术。生物体在养分过量(盛宴)和稀缺性(饥荒)之间处于不同的振荡频率。分析了细胞生理学的变化,例如菌落生长速率和细胞形态,并揭示了不同条件之间生长速率和细胞长度的显着差异。dMSCC还允许应用定义的但随机变化的营养条件,这对于从自然栖息地和大规模生物反应器中复制更复杂的条件非常重要。提出的系统为在复杂多变的环境条件下培养细胞奠定了基础。
更新日期:2020-11-03
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