Research Center for Analytical Sciences, Northeastern University, Shenyang 110819, China.
Talanta. 2011 Apr 15;84(2):565-71. doi: 10.1016/j.talanta.2011.01.065. Epub 2011 Feb 2.
A novel fluid mixing strategy was developed which significantly enhanced the efficiency of DNA hybridization. A pneumatic micro-mixing device consisting of two pneumatic chambers and an underneath DNA microarray chamber was built up. The fluid in the array chamber was pneumatically pumped alternately by the two pneumatic chambers. The chaotic oscillatory flow caused by the pumping greatly intensified the fluidic mixing. A homogeneous distribution of the tracer dye solution in the microarray chamber was observed after 2s mixing with a pumping frequency of 24 Hz. Microarray DNA hybridization was substantially accelerated using this device, and the fluorescence intensity showed a plateau after oscillating 30s at room temperature. The corresponding signal level of the dynamic hybridization was 12.5-fold higher than that of the static hybridization performed at 42°C. A signal-to-noise ratio of 117 was achieved and the nonspecific adsorption of the targets to the sample array was minimized, which might be attributed to the strong shearing force generated during the pneumatic mixing process.
开发了一种新颖的流体混合策略,显著提高了 DNA 杂交的效率。构建了由两个气动腔室和一个下方 DNA 微阵列腔室组成的气动微混合装置。通过两个气动腔室交替泵送,在阵列腔室内产生混沌振荡流,从而极大地增强了流体混合。当泵送频率为 24 Hz 时,在 2 s 混合后观察到微阵列腔室内示踪染料溶液的均匀分布。使用该装置可显著加快微阵列 DNA 杂交,并且在室温下振荡 30 s 后荧光强度达到平台。动态杂交的相应信号水平比在 42°C 下进行的静态杂交高 12.5 倍。实现了 117 的信噪比,并且最大限度地减少了目标对样品阵列的非特异性吸附,这可能归因于气动混合过程中产生的强剪切力。