Department of Bio-industrial Mechatronics Engineering, National Chung Hsing University, No. 250, Kuo Kuang Road, Taichung 402, Taiwan.
Talanta. 2010 Apr 15;81(1-2):228-34. doi: 10.1016/j.talanta.2009.11.062. Epub 2009 Dec 3.
A Clark-type oxygen chip consisting of a polydimethylsiloxane (PDMS) reservoir containing an amino group-modified PDMS oxygen-permeable membrane (OPM) and a glass substrate containing a three-electrode detector has been constructed by using microfabrication techniques, and it is utilized for in situ measurement of the respiration activity of adhering cells. Use of the alginate sol electrolyte and the electroplating Ag/AgCl pseudo-reference electrode can effectively diminish the crosstalk between the electrochemical electrodes and supply a stable potential for the detection of dissolved oxygen, respectively. The Clark-type oxygen chips possess only 1.00% residual current, response time of 13.4s and good linearity with a correlation coefficient of 0.9933. The modification of amino groups for the OPM obviously facilitates the adhesion of HeLa cells onto the PDMS OPM surface and allows the cells to spread after 2h of incubation. The oxygen consumption of the cells in the cell-adhesion process increases with the adhesion time, and the increment of cellular oxygen consumption per minute reaches a maximum after 30 min of incubation. Moreover, the change in the respiration activity of adhering HeLa cells stimulated by the high concentration of glucose or propofol anaesthetic can be monitored in real time with the Clark-type oxygen chip.
采用微加工技术构建了一种由聚二甲基硅氧烷（PDMS）储液器组成的 Clark 型氧芯片，该储液器包含一个氨基修饰的 PDMS 透气膜（OPM）和一个含有三电极检测器的玻璃基底。该氧芯片用于原位测量附着细胞的呼吸活性。使用藻酸盐溶胶电解质和电镀 Ag/AgCl 伪参比电极可以有效减少电化学电极之间的串扰，并为溶解氧的检测提供稳定的电位。Clark 型氧芯片的残余电流仅为 1.00%，响应时间为 13.4s，具有良好的线性关系，相关系数为 0.9933。OPM 的氨基修饰明显促进了 HeLa 细胞在 PDMS OPM 表面的黏附，并允许细胞在孵育 2 小时后铺展。在细胞黏附过程中，细胞的耗氧量随黏附时间的增加而增加，并且在孵育 30 分钟后，每分钟细胞耗氧量的增加达到最大值。此外，Clark 型氧芯片可以实时监测高浓度葡萄糖或丙泊酚麻醉刺激下附着的 HeLa 细胞的呼吸活性变化。
