Trapping and isolation of single prokaryotic cells in a micro-chamber array using dielectrophoresis
Trapping and isolation of single prokaryotic cells in a micro-chamber array using dielectrophoresis
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DOI：
10.1039/c6ra21229h
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发表时间：
2016-01-01
期刊：
影响因子：
3.9
通讯作者：
Yamamoto, T.
中科院分区：
文献类型：
--
作者：
Mogi, K.;Shirataki, C.;Yamamoto, T.
绝大多数原核生物很难或不可能在实验室中培养，这使得使用常规生物化学技术研究这些生物体变得困难。因此，能够物理分离单个原核细胞的方法将通过消除或减少对培养的需要而促进先前未研究的生物体的表征。目前大多数用于单细胞分离的方法被设计用于主要是哺乳动物的真核细胞，其是非运动的并且比原核细胞大得多。因此，我们开发了一种基于微室阵列的方法，用于使用介电泳分离单个原核细胞。在这里，我们证明了该方法的适用性，使用两个原核物种，大肠杆菌（细菌）和Haloferax volcano ii（古细菌），这两个不同的大小和生化组成。我们的研究结果表明，任何一种生物体的细胞被捕获与5至20 MV m（-1）和50 kHz至3 MHz的外加电场，并介电电泳电压和频率的最佳组合取决于细胞类型。我们认为，这种技术是有用的捕获不同的原核物种的单细胞。
The vast majority of prokaryotic species are difficult or impossible to culture in laboratories, which makes it difficult to study these organisms using conventional biochemical techniques. Methods that enable the physical isolation of single prokaryotic cells would thus facilitate the characterization of previously unstudied organisms by eliminating or reducing the need for cultivation. Most current methods for single-cell isolation were designed for eukaryotic cells mainly of mammals, which are non-motile and much larger than prokaryotic cells. We therefore developed a micro-chamber array-based method for the isolation of single prokaryotic cells using dielectrophoresis. Here, we demonstrated the applicability of the method using two prokaryotic species, Escherichia coli (bacteria) and Haloferax volcanii (archaea), which differ both in size and biochemical composition. Our results showed that cells of either organism are trapped with an applied electric field of 5 to 20 MV m(-1) and 50 kHz to 3 MHz, and that the optimum combination of dielectrophoresis voltage and frequency depends on the cell type. We suggest that this technique is useful for trapping single cells of diverse prokaryotic species.