Ultra-smooth glass channels for bioassay with motor proteins
Ultra-smooth glass channels for bioassay with motor proteins
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DOI：
10.1039/b409223f
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发表时间：
2004-01-01
期刊：
影响因子：
4.2
通讯作者：
Fujita, H
中科院分区：
文献类型：
--
作者：
Yokokawa, R;Takeuchi, S;Fujita, H
制作了用于生物分子非荧光观察的光学平板玻璃通道。马达蛋白的两个生物测定成功地进行，其中纳米级珠和微管清楚地观察到的微分干涉对比（DIC）显微镜和暗场显微镜，分别。氢氟酸（HF）的浓度进行了优化，以获得一个光学平坦的表面，通过AFM，SEM和暗场显微镜进行评估。使用聚（二甲基硅氧烷）（PDMS）微流体通道作为掩模蚀刻玻璃通道，并用PDMS涂覆的盖玻片密封。与需要20穆尔的常规流动池方法相比，2-3穆尔的通道体积实现了每次测定所需蛋白质的急剧减少。
Optically-flat glass channels were fabricated for the non-fluorescent observation of bio-molecules. Two bioassays of motor proteins were successfully performed, in which nanoscale beads and microtubules were clearly observed by a differential interference contrast (DIC) microscope and a dark-field microscope, respectively. The concentration of hydrofluoric acid (HF) was optimized to obtain an optically flat surface which was evaluated by AFM, SEM and dark-field microscopy. A glass channel was etched using a poly(dimethyl siloxane) (PDMS) microfluidic channel as a mask and sealed with a PDMS-coated coverslip. The channel volume of 2-3 mul realized the drastic reduction of proteins required per assay compared with a conventional flow cell method requiring 20 mul.