Goto Makiko, Sato Kiichi, Murakami Atsushi, Tokeshi Manabu, Kitamori Takehiko
Department of Applied Chemistry, School of Engineering, University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Anal Chem. 2005 Apr 1;77(7):2125-31. doi: 10.1021/ac040165g.
We developed a novel bioassay system using a glass microchip and cultured cells. A microchamber for cell culture and microchannels for reactions and detection were fabricated on a Pyrex glass substrate by photolithography and wet etching techniques. Cell culture, chemical and enzymatic reactions, and detection were integrated into the microchip. To keep different temperatures locally in three areas of the microchip, we designed and fabricated a temperature control device. Nitric oxide released from macrophage-like cells stimulated by lipopolysaccharide was successfully monitored with the microchip, the temperature control device, and a thermal lens microscope. The total assay time was reduced from 24 to 4 h, and detection limit of NO was improved from 1 x 10(-6) to 7 x 10(-8) M compared with conventional methods. Moreover, the system could monitor a time course of the release, which is difficult to measure by conventional batch methods. We conclude that this system is promising for a rapid bioassay system with very small consumption of cells.
我们开发了一种使用玻璃微芯片和培养细胞的新型生物测定系统。通过光刻和湿蚀刻技术在派热克斯玻璃基板上制造了用于细胞培养的微腔以及用于反应和检测的微通道。细胞培养、化学和酶促反应以及检测被集成到微芯片中。为了在微芯片的三个区域局部保持不同温度，我们设计并制造了一种温度控制装置。利用该微芯片、温度控制装置和热透镜显微镜成功监测了脂多糖刺激的巨噬细胞样细胞释放的一氧化氮。与传统方法相比，总测定时间从24小时缩短至4小时，一氧化氮的检测限从1×10⁻⁶提高到7×10⁻⁸ M。此外，该系统可以监测释放的时间进程，这是传统批量方法难以测量的。我们得出结论，该系统有望成为一种细胞消耗极少的快速生物测定系统。
