Application of optically induced dielectrophoretic (ODEP) force-based microfluidic platform for label-free and high-efficiency live and dead cell separation

• Main Authors: Chien Ting Li, 李建廷
• Other Authors: M. H. Wu
• Format: Others
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/14801371201494513059

碩士 === 長庚大學 === 生化與生醫工程研究所 === 102 === This study reports an optically induced dielectrophoretic (ODEP)force-based microfluidic platform for the separation and collection live and dead cells. The mechanism is based on the fact that live and dead cells have reverse property in under ODEP force field. Under an ODEP background, the live cells are attracted by the applied ODEP force,whereas the dead cells are repelled from the ODEP force. Combining above phenomenon and the flow control in a microfluidic system, the live and dead cells can be separated and subsequently collected in a label-free, efficient and effective manner. In this study, the operating conditions of ODEP force for manipulating the live and dead chondrocytes were first characterized. The performance of live and dead separation was experimentally evaluated. In addition, the impact of ODEP force field on the physiology of the cells manipulated was also investigated. Our results revealed that the applied voltage of 8Vp-p was suitable for the cell separation due to the maximum difference of manipulation force for the live (49.4 pN) and dead (-20.1 pN) cells. In addition, the use of the proposed scheme for live and dead cell separation was successfully demonstrated. We also showed that the recovery rate and purity of the isolated live cells are as high as 78.3±6.8% and 96.4 ±2.2%, respectively. In addition, the purified and isolated live chondrocytes were cultured and observed growth and physiology for eight days. Comparing to electrical unstimulated chondrocytes, experimental cells had no significant difference in cell growth, proliferation and morphology. Also, experimental group and control group have no significant difference in metabolic analysis of lactic acid production and DNA content analysis. In a sum, the proposed cell separation method is found particularly valuable for the biological researches in which the isolation of highly pure live or dead cells is important.