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High purity and viability cell separation of a bacterivorous jakobid flagellate based on a steep velocity gradient induced soft inertial force
Huazhong Univ Sci & Technol, State Key Lab Digital Mfg Equipment & Technol, Wuhan, Hubei, Peoples R China.
Uppsala University, Disciplinary Domain of Science and Technology, Biology, Department of Organismal Biology, Systematic Biology.ORCID iD: 0000-0001-9357-9661
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Microsystems Technology. Huazhong Univ Sci & Technol, State Key Lab Digital Mfg Equipment & Technol, Wuhan, Hubei, Peoples R China.ORCID iD: 0000-0002-3719-406x
2018 (English)In: RSC Advances, ISSN 2046-2069, E-ISSN 2046-2069, Vol. 8, no 62, p. 35512-35520Article in journal (Refereed) Published
Abstract [en]

Cell separation is one of the key limiting factors for precise analysis of non-axenic microbial lab cultures or environmental samples, and it remains a challenge to isolate target cells with high purity and viability via high-throughput cell sorting. During the past decade, hydrodynamic microfluidic platforms have attracted great attention in cell preparation for their high efficiency, robust performance and low cost. Here, we employ the use of a low-velocity sheath flow with high viscosity near the wall and a high-velocity sheath flow with low viscosity on the other side of the sample flow in a soft inertial separation chip. This not only prevents hard interactions between cells and chip walls but, in comparison to previous inertial separation methods, generates a significant increase in deflection of large cells while keeping the small ones in the original flow. We first conducted experiments on a mixture of small and large fluorescent particles (1.0 and 9.9 m, respectively) and removed over 99% of the small particles. The separation efficiency was then tested on a culture of a bacterivorous jakobid flagellate, Seculamonas ecuadoriensis fed on the live bacterium, Klebsiella sp. Using our microfluidic chip, over 94% of live bacteria were removed while maintaining high jakobid cell viability. For comparison, we also conducted size-based cell sorting of the same culture using flow cytometry, which is widely used as a rapid and automated separation tool. Compared with the latter, our chip showed more than 40% higher separation efficiency. Thus, our device provides high purity and viability for cell separation of a sensitive cell sample (jakobid cells). Potentially, the method can be further used for applications in diagnostics, biological analyses and environmental assessment of mixed microbial samples.

Place, publisher, year, edition, pages
ROYAL SOC CHEMISTRY , 2018. Vol. 8, no 62, p. 35512-35520
National Category
Cell Biology Engineering and Technology
Identifiers
URN: urn:nbn:se:uu:diva-369444DOI: 10.1039/c8ra05328fISI: 000448348900021OAI: oai:DiVA.org:uu-369444DiVA, id: diva2:1270741
Funder
Swedish Research Council, 2013-04378Available from: 2018-12-14 Created: 2018-12-14 Last updated: 2019-01-22Bibliographically approved

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Fu, Cheng-JieWu, Zhigang

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