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Design and fabrication of a microfluidic device for near-single cell mRNA isolation using a copper hot embossing master

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Abstract

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 μL. 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.

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Acknowledgments

This work was performed in part at the Cornell NanoScale Facility, a member of the National Nanotechnology Infrastructure Network, which is supported by the National Science Foundation (Grant ECS 03-35765). This work was partially funded under a grant from the New York State Office of Science, Technology and Academic Research (NSYTAR) and a Merck Corporation undergraduate student summer fellowship program in biological engineering. This work also made use of STC shared experimental facilities supported by the National Science Foundation under Agreement No. ECS-9876771.

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  1. Department of Biological and Environmental Engineering, Cornell University, Ithaca, NY, 14853, USA

    S. R. Nugen, P. J. Asiello & A. J. Baeumner

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  1. S. R. Nugen
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  2. P. J. Asiello
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  3. A. J. Baeumner
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Correspondence to A. J. Baeumner.

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Nugen, S.R., Asiello, P.J. & Baeumner, A.J. Design and fabrication of a microfluidic device for near-single cell mRNA isolation using a copper hot embossing master. Microsyst Technol 15, 477–483 (2009). https://doi.org/10.1007/s00542-008-0694-0

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  • DOI: https://doi.org/10.1007/s00542-008-0694-0

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