High-Throughput Single Cell Trapping and Patterning Using a Sandwiched Microfluidic Chip

This paper presents a sandwiched microfluidic chip that allows effective trapping and patterning of single cells using positive-dielectrophoresis. The sandwiched chip consists of a double-layer interdigitated electrode, a thin membrane channel, and a microwell array. Each electrode can be independently controlled to generate spatially different electric fields, and a microwell can only accommodate one cell. After chip design and optimization based on the simulation results of the electric field, a chip prototype was fabricated using the Micro-Electro-Mechanical System technology. The single cell trapping and patterning of this chip were verified experimentally by controlling 97L-GFP and HDFn cells. The single cell trapping efficiency reached 83.7%, indicating that single cells can be trapped into the capture array with high-efficiency. Then, four types of signals were configured, and four single cell patterns were realized, indicating that the trapped single cells can be selectively released for patterning. These experimental results successfully demonstrated that the developed chip can facilitate the study of cell heterogeneity.

[1]  Gabriel P López,et al.  Microfluidic cell sorting: a review of the advances in the separation of cells from debulking to rare cell isolation. , 2015, Lab on a chip.

[2]  David J. Collins,et al.  Two-dimensional single-cell patterning with one cell per well driven by surface acoustic waves , 2015, Nature Communications.

[3]  Gabor Kosa,et al.  Characterization of steady streaming for a particle manipulation system , 2016, Biomedical microdevices.

[4]  Xudong Cao,et al.  Patterning multiple cell types in co-cultures: A review , 2009 .

[5]  Masato Saito,et al.  Single cell trapping and cell–cell interaction monitoring of cardiomyocytes in a designed microfluidic chip , 2015 .

[6]  Dong Sun,et al.  A simplified sheathless cell separation approach using combined gravitational-sedimentation-based prefocusing and dielectrophoretic separation. , 2018, Lab on a chip.

[7]  Yu-Chun Kung,et al.  Microfluidics: Tunnel Dielectrophoresis for Tunable, Single‐Stream Cell Focusing in Physiological Buffers in High‐Speed Microfluidic Flows (Small 32/2016) , 2016 .

[8]  Saeid Nahavandi,et al.  Dielectrophoretic platforms for bio-microfluidic systems. , 2011, Biosensors & bioelectronics.

[9]  Soo Hyeon Kim,et al.  An electroactive microwell array for trapping and lysing single-bacterial cells. , 2011, Biomicrofluidics.

[10]  Marco Bianchessi,et al.  A modular micro-fluidic platform for cells handling by dielectrophoresis , 2010 .

[11]  Gursel Alici,et al.  A review of microfabrication techniques and dielectrophoretic microdevices for particle manipulation and separation , 2014 .

[12]  Robert M Johann,et al.  Cell trapping in microfluidic chips , 2006, Analytical and bioanalytical chemistry.

[13]  A. Folch,et al.  Large-scale single-cell trapping and imaging using microwell arrays. , 2005, Analytical chemistry.

[14]  H K Chu,et al.  Three-dimensional cell manipulation and patterning using dielectrophoresis via a multi-layer scaffold structure. , 2015, Lab on a chip.

[15]  Ming C. Wu,et al.  Massively parallel manipulation of single cells and microparticles using optical images , 2005, Nature.

[16]  Fabien Guillemot,et al.  Cell patterning technologies for organotypic tissue fabrication. , 2011, Trends in biotechnology.

[17]  M. Vellekoop,et al.  Continuous cell from cell separation by traveling wave dielectrophoresis , 2012 .

[18]  Won Gu Lee,et al.  Cell manipulation in microfluidics , 2013, Biofabrication.

[19]  J. Voldman,et al.  A scalable addressable positive-dielectrophoretic cell-sorting array. , 2005, Analytical chemistry.

[20]  Cheng-Hsin Chuang,et al.  Enhancing fluorescent response of immunosensing by a dielectrophoresis chip with transparent electrodes and microcavities array , 2013 .

[21]  R. Pethig Review article-dielectrophoresis: status of the theory, technology, and applications. , 2010, Biomicrofluidics.

[22]  Mitsuhiro Shikida,et al.  Cell culture arrays using magnetic force-based cell patterning for dynamic single cell analysis. , 2008, Lab on a chip.

[23]  Gabor Kosa,et al.  Closed loop control of microscopic particles incorporating steady streaming and visual feedback , 2018, Biomedical microdevices.

[24]  Mina Okochi,et al.  Application of magnetic force‐based cell patterning for controlling cell–cell interactions in angiogenesis , 2009, Biotechnology and bioengineering.

[25]  Shizhi Qian,et al.  Cell electrofusion in microfluidic devices: A review , 2013 .

[26]  Joel Voldman,et al.  nDEP microwells for single-cell patterning in physiological media. , 2007, Lab on a chip.

[27]  Xiao Yan,et al.  Multilevel-Based Topology Design and Cell Patterning With Robotically Controlled Optical Tweezers , 2015, IEEE Transactions on Control Systems Technology.

[28]  Dong Sun,et al.  Enhanced cell sorting and manipulation with combined optical tweezer and microfluidic chip technologies. , 2011, Lab on a chip.

[29]  M. Stelzle,et al.  Microdevices for manipulation and accumulation of micro‐ and nanoparticles by dielectrophoresis , 2003, Electrophoresis.