Electric field directed assembly of high-density microbead arrays.
Electric field directed assembly of high-density microbead arrays.
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DOI：
10.1039/b912876j
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发表时间：
2009-11-21
期刊：
影响因子：
6.1
通讯作者：
Huang X
中科院分区：
文献类型：
--
作者：
Barbee KD;Hsiao AP;Heller MJ;Huang X
我们报告了一种快速、电场定向组装高密度蛋白质共轭微珠阵列的方法。光刻技术用于在涂有一层薄金膜作为主电极的硅片上，在基于环氧基的光致抗蚀剂中制造一系列微米到亚微米级的势垒。薄垫片被用来在晶片和涂有铟-锡氧化物的玻璃盖板之间形成一个微流体室，玻璃盖板用作对电极。将悬浮在低电导缓冲液中的链霉亲和素偶联微球引入电离室，并通过在电极上施加一系列低电压电脉冲，通过电泳将其引导到井中。数亿颗微珠可以在短短30秒内永久组装在这些阵列上，这个过程可以使用荧光显微镜进行实时监控。微珠与金膜的结合很牢固，通过电化学诱导的金-蛋白质相互作用发生，从而允许多余的珠子被洗涤或回收。孔和珠子的大小是这样选择的，即每个孔中只能捕获一个珠子。整个晶片规模阵列的填充效率超过99.9%，密度高达每平方厘米6900万个珠子。这项技术的潜在应用包括组装用于高通量基因组测序的DNA阵列和用于蛋白质组研究的抗体阵列。在阵列组装之后，该装置还可用于通过利用电场加速分子的传输来增强各种分析的浓度依赖过程。
We report a method for rapid, electric field directed assembly of high-density protein-conjugated microbead arrays. Photolithography is used to fabricate an array of micron to sub-micron-scale wells in an epoxy-based photoresist on a silicon wafer coated with a thin gold film, which serves as the primary electrode. A thin gasket is used to form a microfluidic chamber between the wafer and a glass coverslip coated with indium-tin oxide, which serves as the counter electrode. Streptavidin-conjugated microbeads suspended in a low conductance buffer are introduced into the chamber and directed into the wells via electrophoresis by applying a series of low voltage electrical pulses across the electrodes. Hundreds of millions of microbeads can be permanently assembled on these arrays in as little as 30 seconds and the process can be monitored in real time using epifluorescence microscopy. The binding of the microbeads to the gold film is robust and occurs through electrochemically induced gold-protein interactions, which allows excess beads to be washed away or recycled. The well and bead sizes are chosen such that only one bead can be captured in each well. Filling efficiencies greater than 99.9% have been demonstrated across wafer-scale arrays with densities as high as 69 million beads per cm2. Potential applications for this technology include the assembly of DNA arrays for high-throughput genome sequencing and antibody arrays for proteomic studies. Following array assembly, this device may also be used to enhance the concentration-dependent processes of various assays through the accelerated transport of molecules using electric fields.
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影响因子：
56.9
作者：
Shendure, J;Porreca, GJ;Church, GM
通讯作者：
Church, GM
影响因子：
48
作者：
Cloonan, Nicole;Forrest, Alistair R. R.;Grimmond, Sean M.
通讯作者：
Grimmond, Sean M.
影响因子：
4
作者：
Haginoya, C;Ishibashi, M;Koike, K
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Koike, K
影响因子：
4
作者：
Cordovez, Bernardo;Psaltis, Demetri;Erickson, David
通讯作者：
Erickson, David
影响因子：
7.4
作者：
Blicharz, Timothy M.;Siqueira, Walter L.;Helmerhorst, Eva J.;Oppenheim, Frank G.;Wexler, Philip J.;Little, Frederic F.;Walt, David R.
通讯作者：
Walt, David R.