Miniaturized Growth Chambers for the Isolation of "Unculturable" Bacteria

在拟议的项目中，为了改进鹦鹉螺公司目前使用的原位细菌分离系统，将制造并测试一种新设备，以实现单步生长和分离迄今为止一直抵制驯化尝试的新细菌。鹦鹉螺生物科学加拿大公司是一家海洋生物技术公司，专注于发现和开发用于人类和动物健康和保健的海洋天然产品。鹦鹉螺从不同的地理位置分离和培养细菌，从热带到加拿大的北极。这类化合物发现的主要挑战之一是体外培养细胞（通常是细菌）的能力有限。事实上，“大盘子计数异常”表明，只有大约1%的细菌可以在实验室中培养。剩下的99%被认为是“不可培养的”，或者生长缓慢，因为细菌的自然环境中缺乏化学和物理因素。因此，越来越需要开发高通量的原位栽培技术来栽培不可栽培的物种。拟议的微胶囊化室将满足这一需求，并允许更高的培养效率和更广泛的治疗筛选能力。为了解决当前芯片设计的局限性，我们提出了一种新的生长室设计和一种基于微流体的方法，将细菌包封在琼脂糖微珠中。因此，这项研究可能导致分离出新的细菌，产生新的、具有生物活性的天然产物。

In the proposed project, to improve the current in situ bacteria isolation systems used by Nautilus, a noveldevice will be fabricated and tested to enable single-step growth and isolation of new bacteria that have, thusfar, resisted attempts to domesticate. Nautilus Biosciences Canada Inc. is a marine biotechnology companyfocused on the discovery and development of marine-derived natural products with applications in human andanimal health and wellness. Nautilus isolates and cultures bacteria from diverse geographic locations rangingfrom the tropics to Canada's Arctic. One of the main challenges to this type of compound discovery is thelimited ability of culturing cells, usually bacteria, in vitro. In fact, "The Great Plate Count Anomaly" suggeststhat only about 1% of bacteria are cultivable in laboratories. The remaining 99% are considered 'uncultivable',or have slow growth, due to the lack of chemical and physical factors present in the bacteria's naturalenvironments. Therefore, there is an increasing need for development of high throughput in situ technologiesfor cultivation of uncultivable species. The proposed microencapsulation chambers would address this need,and allow for greater culturing efficiency and a broader screening capability for therapeutics. To address thelimitations of the current iChip designs, we propose a new design of a growth chamber and amicrofluidic-based method of encapsulating bacteria in agarose micro-beads. Thus, the research proposedcould lead to the isolation of new bacteria which produce new, bioactive natural products.