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首页> 外文期刊>RSC Advances >High purity and viability cell separation of a bacterivorous jakobid flagellate based on a steep velocity gradient induced soft inertial force
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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

机译：基于陡速度梯度诱导的软惯性，诱导诱导的诱导诱导的诱导噬菌体巨石鞭毛的高纯度和活力细胞分离

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摘要

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.
机译：细胞分离是用于非轴烯微生物实验室培养物或环境样品的精确分析的关键限制因素之一，并且通过高通量细胞分选将靶细胞分离具有高纯度和活力的靶细胞仍然是挑战。在过去十年中，流体动力学微流体平台在细胞准备中引起了巨大的关注，以获得其高效率，稳健的性能和低成本。在这里，我们采用在壁附近的高粘度和高速鞘流的高粘度的使用，并且在软惯性分离芯片中的样品流的另一侧具有低粘度的高速鞘流。这不仅可以防止细胞和芯片壁之间的硬相互作用，而且与先前的惯性分离方法相比，在将小细胞保持在原始流动中的同时产生显着增加。我们首先在小型和大型荧光颗粒（分别为1.0和9.9μm）的混合物上进行实验，并除去超过99％的小颗粒。然后对诱导在活细菌饲喂的诱导抗菌葡萄鞭液的培养物上进行分离效率，Klebsiella sp饲喂eSculamonas ecuadoriensis。使用我们的微流体芯片，在保持高jakobid细胞活力的同时除去超过94％的活细菌。为了比较，我们还使用流式细胞术进行相同培养的基于尺寸的细胞分选，这被广泛用作快速和自动分离工具。与后者相比，我们的芯片显示出超过40％的分离效率。因此，我们的装置提供了敏感细胞样品（Jakobid细胞）的细胞分离的高纯度和活力。潜在地，该方法可以进一步用于诊断中的应用，生物分析和混合微生物样品的环境评估。

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  • 来源
    《RSC Advances》 |2018年第62期|共9页
  • 作者

    Deng Pan; Fu Cheng-Jie; Wu Zhigang;

  • 作者单位

    Huazhong Univ Sci &

    Technol State Key Lab Digital Mfg Equipment &

    Technol Wuhan Hubei Peoples R China;

    Uppsala Univ Dept Organismal Biol Uppsala Sweden;

    Huazhong Univ Sci &

    Technol State Key Lab Digital Mfg Equipment &

    Technol Wuhan Hubei Peoples R China;

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  • 正文语种 eng
  • 中图分类 化学;
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