Lam Raymond H W, Kim Min-Cheol, Thorsen Todd
Department of Mechanical Engineering, Hatsopoulos Microfluids Laboratory, Massachusetts Institute of Technology, Room 3-246, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, USA.
Anal Chem. 2009 Jul 15;81(14):5918-24. doi: 10.1021/ac9006864.
In this manuscript, we report on the culture of anaerobic and aerobic species within a disposable multilayer polydimethylsiloxane (PDMS) microfluidic device with an integrated differential oxygenator. A gas-filled microchannel network functioning as an oxygen-nitrogen mixer generates differential oxygen concentration. By controlling the relative flow rate of the oxygen and nitrogen input gases, the dissolved oxygen (DO) concentration in proximal microchannels filled with culture media are precisely regulated by molecular diffusion. Sensors consisting of an oxygen-sensitive dye embedded in the fluid channels permit dynamic fluorescence-based monitoring of the DO concentration using low-cost light-emitting diodes. To demonstrate the general utility of the platform for both aerobic and anaerobic culture, three bacteria with differential oxygen requirements (E. coli, A. viscosus, and F. nucleatum), as well as a model mammalian cell line (murine embryonic fibroblast cells (3T3)), were cultured. Growth characteristics of the selected species were analyzed as a function of eight discrete DO concentrations, ranging from 0 ppm (anaerobic) to 42 ppm (fully saturated).
在本论文中，我们报道了在具有集成式差分化氧器的一次性多层聚二甲基硅氧烷（PDMS）微流控装置中对厌氧和好氧菌种的培养。一个充当氧 - 氮混合器的充气微通道网络产生不同的氧浓度。通过控制氧气和氮气输入气体的相对流速，充满培养基的近端微通道中的溶解氧（DO）浓度通过分子扩散得到精确调节。由嵌入流体通道的氧敏染料组成的传感器允许使用低成本发光二极管基于动态荧光监测DO浓度。为了证明该平台对好氧和厌氧培养的通用性，培养了三种具有不同氧气需求的细菌（大肠杆菌、粘性放线菌和具核梭杆菌）以及一种模型哺乳动物细胞系（小鼠胚胎成纤维细胞（3T3））。所选菌种的生长特性作为八个离散DO浓度（范围从0 ppm（厌氧）到42 ppm（完全饱和））的函数进行了分析。
