Dang Fuquan, Tabata Osamu, Kurokawa Masaya, Ewis Ashraf A, Zhang Lihua, Yamaoka Yoshihisa, Shinohara Shouji, Shinohara Yasuo, Ishikawa Mitsuru, Baba Yoshinobu
Single-Molecule Bioanalysis Laboratory, National Institute of Advanced Industrial Science and Technology (AIST), Hayashi-cho 2217-14, Takamatsu 761-0395, Japan.
Anal Chem. 2005 Apr 1;77(7):2140-6. doi: 10.1021/ac0485031.
We have developed a novel technique for mass production of microfabricated capillary array electrophoresis (mu-CAE) plastic chips for high-speed, high-throughput genetic analysis. The mu-CAE chips, containing 10 individual separation channels of 50-microm width, 50-microm depth, and a 100-microm lane-to-lane spacing at the detection region and a sacrificial channel network, were fabricated on a poly(methyl methacrylate) substrate by injection molding and then bonded manually using a pressure-sensitive sealing tape within several seconds at room temperature. The conditions for injection molding and bonding were carefully characterized to yield mu-CAE chips with well-defined channel and injection structures. A CCD camera equipped with an image intensifier was used to monitor simultaneously the separation in a 10-channel array with laser-induced fluorescence detection. High-performance electrophoretic separations of phiX174 HaeIII DNA restriction fragments and PCR products related to the human beta-globin gene and SP-B gene (the surfactant protein B) have been demonstrated on mu-CAE plastic chips using a methylcellulose sieving matrix in individual channels. The current work demonstrated greatly simplified the fabrication process as well as a detection scheme for mu-CAE chips and will bring the low-cost mass production and application of mu-CAE plastic chips for genetic analysis.
我们已经开发出一种用于大规模生产微制造毛细管阵列电泳(mu-CAE)塑料芯片的新技术,用于高速、高通量基因分析。mu-CAE芯片包含10个独立的分离通道,通道宽度为50微米,深度为50微米,在检测区域的通道间间距为100微米,还有一个牺牲通道网络。该芯片通过注塑成型在聚甲基丙烯酸甲酯基板上制造,然后在室温下使用压敏密封胶带在几秒钟内手动键合。对注塑成型和键合条件进行了仔细表征,以生产出具有明确通道和进样结构的mu-CAE芯片。配备图像增强器的CCD相机用于通过激光诱导荧光检测同时监测10通道阵列中的分离情况。使用甲基纤维素筛分基质在单个通道中,已在mu-CAE塑料芯片上展示了phiX174 HaeIII DNA限制性片段以及与人β-珠蛋白基因和SP-B基因(表面活性剂蛋白B)相关的PCR产物的高效电泳分离。当前工作极大地简化了mu-CAE芯片的制造过程以及检测方案,并将实现mu-CAE塑料芯片用于基因分析的低成本大规模生产和应用。