Shadpour Hamed, Hupert Mateusz L, Patterson Donald, Liu Changgeng, Galloway Michelle, Stryjewski Wieslaw, Goettert Jost, Soper Steven A
Department of Chemistry, Louisiana State University, Baton Rouge, Louisiana 70803, USA.
Anal Chem. 2007 Feb 1;79(3):870-8. doi: 10.1021/ac0612168.
A 16-channel microfluidic chip with an integrated contact conductivity sensor array is presented. The microfluidic network consisted of 16 separation channels that were hot-embossed into polycarbonate (PC) using a high-precision micromilled metal master. All channels were 40 microm deep and 60 microm wide with an effective separation length of 40 mm. A gold (Au) sensor array was lithographically patterned onto a PC cover plate and assembled to the fluidic chip via thermal bonding in such a way that a pair of Au microelectrodes (60 microm wide with a 5 microm spacing) was incorporated into each of the 16 channels and served as independent contact conductivity detectors. The spacing between the corresponding fluidic reservoirs for each separation channel was set to 9 mm, which allowed for loading samples and buffers to all 40 reservoirs situated on the microchip in only five pipetting steps using an 8-channel pipettor. A printed circuit board (PCB) with platinum (Pt) wires was used to distribute the electrophoresis high-voltage to all reservoirs situated on the fluidic chip. Another PCB was used for collecting the conductivity signals from the patterned Au microelectrodes. The device performance was evaluated using microchip capillary zone electrophoresis (mu-CZE) of amino acid, peptide, and protein mixtures as well as oligonucleotides that were separated via microchip capillary electrochromatography (mu-CEC). The separations were performed with an electric field (E) of 90 V/cm and were completed in less than 4 min in all cases. The conductivity detection was carried out using a bipolar pulse voltage waveform with a pulse amplitude of +/-0.6 V and a frequency of 6.0 kHz. The conductivity sensor array concentration limit of detection (SNR = 3) was determined to be 7.1 microM for alanine. The separation efficiency was found to be 6.4 x 10(4), 2.0 x 10(3), 4.8 x 10(3), and 3.4 x 10(2) plates for the mu-CEC of the oligonucleotides and mu-CZE of the amino acids, peptides, and proteins, respectively, with an average channel-to-channel migration time reproducibility of 2.8%. The average resolution obtained for mu-CEC of the oligonucleotides and mu-CZE of the amino acids, peptides, and proteins was 4.6, 1.0, 0.9, and 1.0, respectively. To the best of our knowledge, this report is the first to describe a multichannel microchip electrophoresis device with integrated contact conductivity sensor array.
本文介绍了一种集成了接触式电导传感器阵列的16通道微流控芯片。微流控网络由16个分离通道组成,这些通道通过高精度微铣金属母模热压印到聚碳酸酯(PC)中。所有通道深度为40微米,宽度为60微米,有效分离长度为40毫米。金(Au)传感器阵列通过光刻图案化在PC盖板上,并通过热键合组装到流体芯片上,使得16个通道中的每一个都包含一对Au微电极(宽度为60微米,间距为5微米),并用作独立的接触式电导检测器。每个分离通道相应流体储液器之间的间距设置为9毫米,这使得使用8通道移液器仅通过五步移液操作就能将样品和缓冲液加载到微芯片上的所有40个储液器中。一个带有铂(Pt)线的印刷电路板(PCB)用于将电泳高压分配到位于流体芯片上的所有储液器。另一个PCB用于收集来自图案化Au微电极的电导信号。使用氨基酸、肽和蛋白质混合物以及通过微芯片毛细管电色谱(mu-CEC)分离的寡核苷酸的微芯片毛细管区带电泳(mu-CZE)对该设备性能进行了评估。分离在90 V/cm的电场(E)下进行,所有情况下均在不到4分钟内完成。使用双极脉冲电压波形进行电导检测,脉冲幅度为+/-0.6 V,频率为6.0 kHz。丙氨酸的电导传感器阵列检测限(SNR = 3)确定为7.1 microM。寡核苷酸的mu-CEC以及氨基酸、肽和蛋白质的mu-CZE的分离效率分别为6.4×10⁴、2.0×10³、4.8×10³和3.4×10²理论塔板数,通道间迁移时间的平均重现性为2.8%。寡核苷酸的mu-CEC以及氨基酸、肽和蛋白质的mu-CZE获得的平均分辨率分别为4.6、1.0、0.9和1.0。据我们所知,本报告首次描述了一种具有集成接触式电导传感器阵列的多通道微芯片电泳设备。