title: Continuous nucleus extraction by optically-induced cell lysis on a batch-type microfluidic platform.
abstract: The extraction of a cell's nucleus is an essential technique required for a number of procedures, such as disease diagnosis, genetic replication, and animal cloning. However, existing nucleus extraction techniques are relatively inefficient and labor-intensive. Therefore, this study presents an innovative, microfluidics-based approach featuring optically-induced cell lysis (OICL) for nucleus extraction and collection in an automatic format. In comparison to previous micro-devices designed for nucleus extraction, the new OICL device designed herein is superior in terms of flexibility, selectivity, and efficiency. To facilitate this OICL module for continuous nucleus extraction, we further integrated an optically-induced dielectrophoresis (ODEP) module with the OICL device within the microfluidic chip. This on-chip integration circumvents the need for highly trained personnel and expensive, cumbersome equipment. Specifically, this microfluidic system automates four steps by 1) automatically focusing and transporting cells, 2) releasing the nuclei on the OICL module, 3) isolating the nuclei on the ODEP module, and 4) collecting the nuclei in the outlet chamber. The efficiency of cell membrane lysis and the ODEP nucleus separation was measured to be 78.04 ± 5.70% and 80.90 ± 5.98%, respectively, leading to an overall nucleus extraction efficiency of 58.21 ± 2.21%. These results demonstrate that this microfluidics-based system can successfully perform nucleus extraction, and the integrated platform is therefore promising in cell fusion technology with the goal of achieving genetic replication, or even animal cloning, in the near future.
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Republic of China. gwobin@pme.nthu.edu.tw.
authors.forename: Shih-Hsuan
authors.initials: SH
authors.lastname: Huang
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Republic of China. gwobin@pme.nthu.edu.tw.
authors.forename: Lien-Yu
authors.initials: LY
authors.lastname: Hung
authors.affiliation: Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Republic of China. gwobin@pme.nthu.edu.tw and Institute of Biomedical Engineering, National Tsing Hua University, Hsinchu 30013, Republic of China and Institute of NanoEngineering and Microsystems, National Tsing Hua University, Hsinchu 30013, Republic of China.
authors.forename: Gwo-Bin
authors.initials: GB
authors.lastname: Lee
mesh_terms: D016923:Cell Death; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus; D004560:Electricity; D004867:Equipment Design; D057809:HEK293 Cells; D006801:Humans; D056656:Lab-On-A-Chip Devices; D055620:Optical Phenomena