Micropreparative fraction collection in microfluidic devices
Micropreparative fraction collection in microfluidic devices
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DOI：
10.1021/ac0112364
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发表时间：
2002-04-01
影响因子：
7.4
通讯作者：
Guttman, A
中科院分区：
文献类型：
--
作者：
Khandurina, J;Chován, T;Guttman, A
基于微芯片的生物分子电泳分析后的微制备级分收集对于各种生物医学应用具有重要意义。在本文中，我们提出了一个微加工设备为基础的馏分收集系统。通过使用适当的电压操作简单地将检测的样品区的所需部分重新定向到相应的收集威尔斯孔，分离和收集各种大小的DNA片段。通过在检测点处或检测点下游放置交叉通道，提高了级分的采样和收集效率。在检测到感兴趣的条带之后，将电位重新配置为采样/收集模式，使得所选择的样品区迁移到微型器件的适当收集孔。电位分布确保了分离柱中剩余的分析物组分被延迟、停止或反转，以这种方式增加了正在收集的样品区与紧接着的样品区之间的间隔。通过这种方式，可以促进空间上接近的连续带的精确收集。一旦目标样品级分到达相应的收集孔，则将电位切换回分离模式。重复交替分离/检测和采样/收集循环，直至所有所需的样品区被物理隔离。该集成装置由进样、分离、馏分取样和馏分收集隔室组成。在dsDNA片段的混合物上测试了级分收集技术的可行性。以这种方式收集的DNA量足以用于进一步的下游样品处理，例如常规的基于PCR的分析。
Micropreparative fraction collection following microchip-based electrophoretic analysis of biomolecules is of major importance for a variety of biomedical applications. In this paper, we present a microfabricated device-based fraction collection system. Various size DNA fragments were separated and collected by simply redirecting the desired portions of the detected sample zones to corresponding collection wells using appropriate voltage manipulations. The efficiency of sampling and collection of the fractions was enhanced by placing a cross channel at or downstream of the detection point. Following the detection of the band of interest, the potentials were reconfigured to sampling/collection mode, so that the selected sample zone migrated to the appropriate collection well of the microdevice. The potential distribution assured that the rest of the analyte components in the separation column was retarded, stopped, or reversed, increasing in this way the spacing between the sample zone being collected and the immediately following one. By this means, a precise collection of spatially close consecutive bands could be facilitated. Once the target sample fraction reached the corresponding collection well, the potentials were switched back to separation mode. Alternation of the separation/ detection and sampling/collection cycles was repeated until all required sample zones were physically isolated. The integrated device consists of a sample introduction, separation, fraction sampling, and fraction collection compartments. The feasibility of the fraction collection technique was tested on a mixture of dsDNA fragments. The amounts of DNA collected in this way were enough for further downstream sample processing, such as conventional PCR-based analysis.