Separation and concentration of Phytophthora ramorum sporangia by inertial focusing in curving microfluidic flows
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- 作者：Liviu Clime ; Kebin Li ; Matthias Geissler ; Xuyen D. Hoa…
- 关键词：Inertial separation ; Microfluidic chip ; Plant pathogen detection ; Pneumatic pumping
- 刊名：Microfluidics and Nanofluidics
- 出版年：2017
- 出版时间：January 2017
- 年：2017
- 卷：21
- 期：1
- 全文大小：
- 刊物类别：Engineering
- 刊物主题：Engineering Fluid Dynamics; Biomedical Engineering; Analytical Chemistry; Nanotechnology and Microengineering;
- 出版者：Springer Berlin Heidelberg
- ISSN：1613-4990
- 卷排序：21
文摘
We present an integrated microfluidic system for performing isolation and concentration of Phytophthora ramorum pathogens using a chip whose working principle is based on inertial lateral migration in curving flows. The chip was fabricated from multiple layers of thermoplastic polymers and features an embedded spiral separation channel along with peristaltic microvalves for fluidic operation and process control. A pumping system paired with a fully programmable pressure manifold is used to boost concentration levels by recirculating the sample liquid multiple times through the separation chip, making it possible to reduce sample volumes from 10 to 1 mL or less. The system was calibrated using fluorescent polymer particles with a nominal diameter of 30 µm which is comparable to that of P. ramorum sporangia. The separation process has been shown to be highly effective and more than 99% of the beads can be recovered in the concentrated batch. Experiments conducted with P. ramorum sporangia have shown that a 5.3-fold increase in pathogen content with 95% recovery can be achieved using three subsequent concentration cycles. The utility of the method has been validated by processing a sample derived from infested Rhododendron leaves where a 6.1-fold increase in the concentration of P. ramorum has been obtained after four concentration cycles. Although specifically designed and demonstrated for sporangia of P. ramorum, the method and related design rules can easily be extended to other microbial organisms, effectively supporting bioanalytical applications where efficient, high-throughput separation of target species is of primary concern.