Center for Microfluidics and Nanotechnology and School of Physics, The State Key Laboratory for Artificial Microstructures and Mesoscopic Physics, Peking University, Beijing, 100871, China.
Biomed Microdevices. 2010 Jun;12(3):499-503. doi: 10.1007/s10544-010-9406-5.
We have developed a microturbidostat for long time bacterial culture at constant density controlled by optical detection and integrated pneumatic valves. The device was fabricated by multilayer soft lithography and in-situ formation of an agarose filter. The culture chamber of bacteria was connected in one side to a single bacterial input-output channel and in another side to a nutrient channel in which the agarose filter was formed to ensure the diffusion of nutrients and metabolites without bacterial loss. The bacterial number in the culture chamber was determined by measuring the fluorescence intensity of GFP proteins of the bacteria and the redundant bacteria could be exported automatically through the input-output channel with integrated micro-valves. In order to optimize the operation performance, we investigated the bacterial exportation efficiency with different input-output channel widths. As expected, the bacterial sorting coefficient was proportional to the input-output channel width. The results also showed that with a 20 microm channel-width, a long time culture was possible with a constant bacterial number in the chamber in the range from 400 to 700.
我们开发了一种微浊度计，用于通过光学检测和集成气动阀控制的恒密度长时间细菌培养。该设备通过多层软光刻和琼脂糖过滤器的原位形成制造。细菌培养室的一侧连接到单个细菌输入-输出通道，另一侧连接到形成琼脂糖过滤器的营养物通道，以确保营养物和代谢物的扩散而不会导致细菌损失。通过集成微阀的输入-输出通道，可以自动排出多余的细菌，并通过测量细菌 GFP 蛋白的荧光强度来确定培养室内的细菌数量。为了优化操作性能，我们研究了不同输入-输出通道宽度下的细菌排出效率。正如预期的那样，细菌分拣系数与输入-输出通道宽度成正比。结果还表明，在 20 微米通道宽度下，在 400 到 700 范围内，培养室内的细菌数量可以保持恒定，从而实现长时间培养。
