Design and fabrication of a microfluidic device for near-single cell mRNA isolation using a copper hot embossing master
Design and fabrication of a microfluidic device for near-single cell mRNA isolation using a copper hot embossing master
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DOI：
10.1007/s00542-008-0694-0
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发表时间：
2009-03-01
影响因子：
2.1
通讯作者：
Baeumner, A. J.
中科院分区：
文献类型：
--
作者：
Nugen, S. R.;Asiello, P. J.;Baeumner, A. J.
我们描述了一种一次性聚合物基芯片的真核mRNA的分离的调查。本工作的重点是改进快速成型的制造方法和在最低RNA浓度下的实际应用，总通道体积为3.5 μ L。使用微流体通道内的顺磁性寡聚（dT）（25）珠实现信使RNA分离，所述微流体通道并入微结构化设计以帮助混合。与没有结构的通道相比，所述结构被示出为促进平行流中的两种流体之间的混合。该芯片是通过热压聚（甲基丙烯酸甲酯）（PMMA）使用铜母版。由于铜具有优异的热、机械和电镀性能，因此被用作母材。母版的制造包括使用KMPR 1050作为用于形成微通道结构的电镀模具对抛光铜板进行结构化。发现铜母版比用于原型制作的传统硅母版更坚固。KMPR的使用使得能够生成与SU-8母版相反的高直壁。此外，由于改善的热传导和避免了冗长和精细的去压花步骤，压花时间能够减少3倍。
We describe investigations toward a disposable polymer-based chip for the isolation of eukaryotic mRNA. This work focuses here on the improvement of the fabrication methods for rapid prototyping and the actual application at lowest RNA concentrations with total channel volumes of 3.5 mu L. Messenger RNA isolation was achieved using paramagnetic oligo (dT)(25) beads within a microfluidic channel which incorporated a sawtooth microstructured design to aid in mixing. The structures were shown to facilitate mixing beteen two fluids in parallel flow when compared to a channel without structures. The chip was fabricated by means of hot embossing poly(methyl methacrylate) (PMMA) using a copper master. Copper was used as the master material due to its excellent thermal, mechanical, and electroplating properties. Fabrication of the master consisted of the structuring of a polished copper plate using KMPR 1050 as an electroplating mold for forming the microchannel structures. The copper master was found to be much more robust than traditional silicon masters used for prototyping. The use of KMPR enabled the generation of high straight walls in contrast to SU-8 masters. In addition, embossing times were able to be decreased by a factor of 3 due to improved heat conduction and avoidance of a lengthy and delicate de-embossing step.