Panaro Nicholas J, Lou Xing Jian, Fortina Paolo, Kricka Larry J, Wilding Peter
Department of Pathology and Laboratory Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Biomol Eng. 2005 Feb;21(6):157-62. doi: 10.1016/j.bioeng.2004.11.001.
We report the fabrication of silicon chips containing a row of 667 pillars, 10 by 20 microm in cross-section, etched to a depth of 80 microm with adjacent pillars being separated by 3.5 microm. The chips were used to separate white blood cells from whole blood in less than 2 min and for subsequent PCR of a genomic target (eNOS). Chip fluid dynamics were validated experimentally using CoventorWare microfluidic simulation software. The amplicon concentrations were determined using microchip capillary electrophoresis and were >40% of that observed in conventional PCR tubes for chips with and without pillars. Reproducible on-chip PCR was achieved using white blood cell preparations isolated from whole human blood pumped through the chip.
我们报告了含有一排667个柱子的硅芯片的制造，这些柱子的横截面为10×20微米，蚀刻深度为80微米，相邻柱子之间的间距为3.5微米。这些芯片用于在不到2分钟的时间内从全血中分离白细胞，并用于随后对基因组靶点（内皮型一氧化氮合酶）进行聚合酶链反应（PCR）。使用CoventorWare微流体模拟软件通过实验验证了芯片的流体动力学。使用微芯片毛细管电泳测定扩增子浓度，对于有柱子和没有柱子的芯片，其扩增子浓度均大于在传统PCR管中观察到的浓度的40%。使用从通过芯片泵送的全人类血液中分离的白细胞制剂实现了可重复的芯片上PCR。
