Park Jaehyun, Bansal Tushar, Pinelis Mikhail, Maharbiz Michel M
Electrical Engineering and Computer Science Department, University of Michigan, Ann Arbor, 1301 Beal Ave., Ann Arbor, MI 48109-2122, USA.
Lab Chip. 2006 May;6(5):611-22. doi: 10.1039/b516483d. Epub 2006 Mar 16.
We present the design, modeling, fabrication and testing of a microsystem for the electrolytic patterning and sensing of oxidative microgradients within 1 x 1 mm2 area during cell culture. The system employs an array of microfabricated electrodes (3-40 microm in width) embedded in gas-permeable microchannels to generate precise doses of dissolved oxygen (ranging from 10 fmol O2 mm(-2) s(-1) to 100 nmol O2 mm(-2) s(-1)) via electrolysis. The microgradients generated by different microelectrodes in the array can be superimposed to pattern multi-dimensional oxygen profiles not possible with other methods. We demonstrate the patterning, sensing and quantification of dissolved oxygen microgradients in the 0 to 40% dO2 range using this microsystem. Reactive oxygen species generation and dosing is also quantified. Lastly, we demonstrate how the microtechnology enables new types of experiments in three different cell culture models: localized hyperoxia-induced apoptosis in C2C12 myoblasts, dynamic aerotaxis assays of Bacillus subtilis, and studies of calcium release in an ischemia/re-oxygenation myoblast model.
我们展示了一种微系统的设计、建模、制造和测试，该微系统用于在细胞培养过程中对1×1平方毫米区域内的氧化微梯度进行电解图案化和传感。该系统采用嵌入透气微通道的微制造电极阵列（宽度为3 - 40微米），通过电解产生精确剂量的溶解氧（范围从10飞摩尔O₂ 毫米⁻² 秒⁻¹ 到100纳摩尔O₂ 毫米⁻² 秒⁻¹）。阵列中不同微电极产生的微梯度可以叠加，以形成其他方法无法实现的多维氧分布图案。我们使用该微系统展示了在0至40%溶解氧范围内溶解氧微梯度的图案化、传感和定量。还对活性氧的产生和剂量进行了定量。最后，我们展示了该微技术如何在三种不同的细胞培养模型中实现新型实验：C2C12成肌细胞中局部高氧诱导的细胞凋亡、枯草芽孢杆菌的动态趋氧性测定以及缺血/再氧合成肌细胞模型中的钙释放研究。
