Analytical Sciences

Simple Isolation of Single Cell: Thin Glass Microfluidic Device for Observation of Isolated Single Euglena gracilis Cells

Nobutoshi OTA,* Yaxiaer YALIKUN,*,** Nobuyuki TANAKA,* Yigang SHEN,* Yusufu AISHAN,* Yuki NAGAHAMA,*** Minoru OIKAWA,*** and Yo TANAKA*

*Center for Biosystems Dynamics Research, RIKEN, 1-3 Yamadaoka, Suita, Osaka 565-0871, Japan
**Division of Materials Science, Nara Institute of Science and Technology, Takayama, Ikoma, Nara 630-0192, Japan
***Department of Engineering, Graduate School of Engineering, Chiba University, 1-33 Yayoi, Inage, Chiba, Chiba 263-8522, Japan

Single cell analysis has gained attention as a means to investigate the heterogeneity of cells and amplify a cell with desired characteristics. However, obtaining a single cell from a large number of cells remains difficult because preparation of single-cell samples relies on conventional techniques such as pipetting that are labor intensive. In this study, we developed a system combining a 0.6-mm thin glass microfluidic device and machine vision approach to isolate single Euglena gracilis cells, as a model of microorganism with mobility, in a small/thin glass chamber. A single E. gracilis cell in a chamber was cultured for 4 days to monitor its multiplication. With this system, we successfully simplified preparation of single cells of interest and determined that it is possible to combine it with other analytical techniques to observe single cells continuously.

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