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MONOLITHIC 3-D MICROFLUIDIC DEVICE FOR CELL ASSAY WITH AN INTEGRATED COMBINATORIAL MIXER

MONOLITHIC 3-D MICROFLUIDIC DEVICE FOR CELL ASSAY WITH AN INTEGRATED COMBINATORIAL MIXER. Mike C. Liu, Dean Ho, Yu-Chong Tai. Department of Bioengineering, California Institute of Technology, Pasadena, USA.

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MONOLITHIC 3-D MICROFLUIDIC DEVICE FOR CELL ASSAY WITH AN INTEGRATED COMBINATORIAL MIXER

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  1. MONOLITHIC 3-D MICROFLUIDIC DEVICE FOR CELL ASSAY WITH AN INTEGRATED COMBINATORIAL MIXER Mike C. Liu, Dean Ho, Yu-Chong Tai Department of Bioengineering, California Institute of Technology, Pasadena, USA Department of Biomedical and Mechanical Engineering, Northwestern University, Evanston, USA Department of Electrical Engineering, California Institute of Technology, Pasadena, USA Transducers’07 pp.787-790 陳睿鈞

  2. Outline • Introduction • Device design and fabrication • Experimental and discussion • Conclusion

  3. Outline • Introduction • Device design and fabrication • Experimental and discussion • Conclusion

  4. Biological Assays Devices • Drug screening and biological assays often include multiple combinations of different compounds. Traditional screening tools Microfluidic devices robotics multi-well plates P. J. Lee, 2006 K. R. King,2007 T. Chapman, 2003 Shane J. Stafslien, 2005 Poor small-volume liquid handling ability Large consumption of reagents High cost of operation Inexpensive chip-platforms High-density arrays Only expose cells to a single compound at once

  5. 3-D Microfluidic Combinatorial Mixer Combinatorial Mixer Individually chamber LOC device Streams control

  6. Outline • Introduction • Device design and fabrication • Experimental and discussion • Conclusion

  7. Design Overpass Allow one microfluidic channel to cross over other microfluidic channels 1 cm×1 cm chip Combinatorial mixer Deliver different solution combinations to the culture-chambers Three inputs seven possible outputs One control channel Cell culture-chambers Cells culture

  8. Device Fabrication 1.Si wafer clean : H2SO4:H2O2 = 3:1 Promote adhesion : DI water:IPA:A-174 = 100:100:1 2.Parylene-coated Si : 3μm Sacrificial photoresist AZ4620 : 15μm Parylene : 10μm 3.Pattern parylene : oxygen plasma 4.Sacrificial photoresist AZ4620 : 32μm Parylene : 10μm 5.SU-8 : 100μm Elute AZ4620 : IPA

  9. Packaging Appliance Transparent acrylic Milled with a computer-numerical controlled (CNC) machine PDMS layer 1. Gasket layer to provide proper sealing 2. Adapter to connect the tubes 3. Adjusted as open or blocked Teflon tubes Plugged into the holes of the PDMS layer Programmable syringe pumps Controll the food coloring solutions load and the flow rate

  10. Outline • Introduction • Device design and fabrication • Experimental and discussion • Conclusion

  11. Combinatorial Mixer Operated flow rate : 0.1L min−1 flow rate : 10L min−1 D : diffusion coefficient U : fluid velocity w: channel width Z : distance during time period

  12. Microfluidic Cell Culture 1.UV irradiation 70% ethanol solution PBS solution 0.05% polyethyleneimine (PEI) : 24h 2.B35 cells adhered to the culture-chamber : 4 h 3.Continuous perfusion of culture media at flowrate of 33 nL/min , 37°C. The cells were grown with continuous perfusion of culture media and pictures were taken 4 h, 16 h and 42 h after cells were loaded.

  13. Simple Cell Assay 1.B35 cells injected 4 h. 2.Injecting 3 cell stains : crystal violet, methylene blue, neutral red. 3.Thecombinatorial mixer 4.The various combinatorial streams into the cell culture-chambers. 5.Cells were stained with different color patterns

  14. Conclusion • The ability to simultaneously treat arrays of cells with different combinations of compounds. • The fruition of such system will enable LOC devices to perform highly parallel and combinatorial chemical or biochemical reactions with reduced labors, reagents and time. • The fabrication technology can enhance the functionalities of current LOC devices by integrating the devices with complex 3-D microfluidic networks. • Future work Monitoring cell growth, more complicated cellular response Real-time monitoring of gene expression

  15. References • Mike C. Liu , Dean Ho, Yu-Chong Tai, “Monolithic fabrication of three-dimensional microfluidic networks forconstructing cell culture array with an integrated combinatorial mixer”, Sensors and Actuators B, 2007. • P. J. Lee, P. J. Hung, V. M. Rao and L. P. Lee, “Nanoliter scale microbioreactor array for quantitative cell biology,” Biotechnology and Bioengineering, Vol. 94, No. 1, pp. 5-14, 2006. • K. R. King, S. Wang, D. Irimia, A. Jayaraman, M. Toner and M. L. Yarmush, “A high-throughput microfluidic real-time gene expression living cell array,” Lab on a Chip, Vol. 7, pp. 77-85, 2007. • T. Chapman, Lab automation and robotics: automation on the move, Nature 421 (2003) 661–666.

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