Isolation of single motile cells using a high-speed picoliter pipette
Isolation of single motile cells using a high-speed picoliter pipette
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DOI：
10.1007/s10404-018-2183-9
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发表时间：
2019-02-01
影响因子：
2.8
通讯作者：
Arai, Fumihito
中科院分区：
文献类型：
--
作者：
Kasai, Yusuke;Sakuma, Shinya;Arai, Fumihito
单细胞的分离是单细胞分析的基本技术。使用单细胞水平的微小培养基将目标细胞从细胞群中分离出来需要高分辨率的受控体积。此外，分离速度也是捕获运动细胞的重要要求。为了实现高速、高分辨率的细胞分离，我们提出了一种高速皮升移液器作为分离装置，它由压电致动器驱动。我们评估了所开发的皮升移液器的性能。结果显示，高分辨率和高速响应分别为 0.42 pL 和 3.9 ms。此外，我们使用皮升移液器分离了细小眼虫（E. gracilis）细胞，它是一种运动细胞。利用发达的皮升移液器的快速响应，我们成功地拾取了一个游泳的单个细小桉细胞。挑取后，将挑取的细胞单独分离到其他培养皿中。我们评估了分离的 E. gracilis 细胞的活力，99.1%（总计：110 个细胞）的细胞显示出运动和分裂能力。根据这些结果，我们实现了用于单细胞分离的高速皮升移液。
Isolation of single cells is a fundamental technique for single-cell analyses. High resolution of controlled volume is required to isolate the target cell from a cell group with tiny medium of single-cell level. Furthermore, the speed of isolation is also an important requirement to capture a motile cell. To achieve cell isolation with high resolution at high speed, we present a high-speed picoliter pipette as an isolation device, which is driven by a piezoelectric actuator. We evaluated performances of the developed picoliter pipette. The results showed a high-resolution and high-speed response of 0.42 pL and 3.9 ms, respectively. In addition, we applied the picoliter pipette to isolate the Euglena gracilis (E. gracilis) cell which was a motile cell. Utilizing the fast response of developed picoliter pipette, we succeeded in picking up a swimming single E. gracilis cell. After the picking up, we isolated the picked cells into other culture dish individually. We evaluated viability of the isolated E. gracilis cells, and 99.1 % (total: 110 cells) of the cells showed motility and division ability. From these results, we achieved the high-speed picoliter pipetting for single-cell isolation.