High purity and viability cell separation of a bacterivorous jakobid flagellate based on a steep velocity gradient induced soft inertial force.
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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基于陡峭速度梯度诱导软惯性力的食菌雅各布氏鞭毛虫的高纯度和活力细胞分离
DOI：
10.1039/c8ra05328f
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发表时间：
2018-10-15
期刊：
影响因子：
3.9
通讯作者：
Wu, Zhigang
中科院分区：
文献类型：
--
作者：
Deng, Pan;Fu, Cheng-Jie;Wu, Zhigang
细胞分离是非无菌微生物实验室培养物或环境样品精确分析的关键限制因素之一，通过高通量细胞分选分离出高纯度和高活性的靶细胞仍然是一个挑战。在过去的十年里，流体动力微流控平台以其高效、坚固的性能和低成本在细胞制备方面引起了人们的极大关注。这里，我们在软惯性分离芯片中使用了靠近壁面的低速度高粘度鞘流和样品流另一侧的低粘度高速鞘流。这不仅防止了细胞和芯片壁之间的硬相互作用，而且与以前的惯性分离方法相比，在保持小细胞保持原始流动的同时，大大增加了大细胞的偏转。我们首先在小颗粒和大颗粒(分别为1.0和9.9μm)的混合物上进行了实验，去除了99%以上的小颗粒。然后在以活细菌克雷伯氏菌为食的雅各布鞭毛虫Seculamonas ecuadoriensis的培养上测试了分离效率。使用我们的微流控芯片，在保持Jakobid细胞高活性的同时，超过94%的活细菌被去除。为了进行比较，我们还使用流式细胞术对同一培养物进行了基于大小的细胞分选，这是一种广泛使用的快速和自动分离工具。与后者相比，该芯片的分离效率提高了40%以上。因此，我们的设备为敏感细胞样本(雅各布细胞)的细胞分离提供了高纯度和高活性。该方法有可能进一步应用于混合微生物样品的诊断、生物分析和环境评价。为了从非无菌微生物实验室培养物或环境样品中分离出高纯度、高活性的活细胞，我们开发了一种操作简单、效率高、性能稳定的新型微流控分离技术。
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 μm, 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. Aimed at separating living cells with high purity and viability from non-axenic microbial lab cultures or environmental samples, we developed a novel microfluidic separation technique with simple operation, high efficiency and robust performance.
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DOI：
10.1007/10_2007_069
发表时间：
2007-01-01
期刊：
CELL SEPARATION: FUNDAMENTALS, ANALYTICAL AND PREPARATIVE METHODS
影响因子：
--
作者：
Dainiak, Maria B.;Kumar, Ashok;Mattiasson, Bo
通讯作者：
Mattiasson, Bo
DOI：
10.1038/nrm4041
发表时间：
2015-09
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Duncombe TA;Tentori AM;Herr AE
通讯作者：
Herr AE
影响因子：
6.2
作者：
Chen, Xing;Cui, Dafu;Chen, Jian
通讯作者：
Chen, Jian
影响因子：
6.1
作者：
Kuntaegowdanahalli, Sathyakumar S.;Bhagat, Ali Asgar S.;Papautsky, Ian
通讯作者：
Papautsky, Ian
影响因子：
6.1
作者：
Li, Youlan;Dalton, Colin;Kaler, Karan V. I. S.
通讯作者：
Kaler, Karan V. I. S.