Matsubara Yasutaka, Kobayashi Masaaki, Morita Yasutaka, Tamiiya Eiichi
The School of Materials Science, Japan Advanced Institute of Science and Technology, Tatsunokuchi, Ishikawa, Japan.
Arch Histol Cytol. 2002 Dec;65(5):481-8. doi: 10.1679/aohc.65.481.
We recently developed a microchamber array chip for DNA amplification by adopting semiconductor microfabrication technology; a polymerase chain reaction (PCR) was performed in the microchamber array, and the amplified DNA was detected using a fluorescent dye. In order to manipulate a single cell or sample into each microchamber individually in this system, the chip was directly sealed with a cover glass slip which impeded the retrieval of the products from each chamber. The present study was therefore carried out to improve the system by developing methods for covering the microchambers and introducing the reaction solution. First, we fabricated a microchamber array chip, and the oil layer was coated on the whole chip instead of the cover glass slip. The solution for DNA amplification was introduced into each chamber through an oil layer using a nano-liter dispenser. Following this, the microarray chip was placed onto the thermal cycling system for DNA amplification, and the amplified DNA was subsequently detected by fluorescence microscopy. In this system, the products were easily retrieved using a micromanipulator for further analysis.
我们最近采用半导体微加工技术开发了一种用于DNA扩增的微腔阵列芯片；在微腔阵列中进行聚合酶链反应（PCR），并使用荧光染料检测扩增的DNA。为了在该系统中分别将单个细胞或样品单独放入每个微腔中，芯片直接用盖玻片密封，这阻碍了从每个腔室中取出产物。因此，本研究旨在通过开发覆盖微腔和引入反应溶液的方法来改进该系统。首先，我们制造了一个微腔阵列芯片，用油层代替盖玻片覆盖整个芯片。使用纳升分配器通过油层将DNA扩增溶液引入每个腔室。随后，将微阵列芯片置于用于DNA扩增的热循环系统上，随后通过荧光显微镜检测扩增的DNA。在该系统中，使用显微操作器很容易取出产物以进行进一步分析。
