Continuous Cell Electroporation for Efficient DNA and siRNA Delivery Based on Laminar Microfluidic Chips

  • Zewen Wei
  • Zhihong Li
Part of the Methods in Molecular Biology book series (MIMB, volume 1121)


Electroporation is a high-efficiency and low-toxicity physical gene transfer method. Traditional electroporation is limited to only low volume cell samples. Here we present a continuous cell electroporation method based on commonly used microfluidic chip fabrication technology. Using easily fabricated PDMS microfluidic chip, syringe pumps, and pulse generator, we show efficient delivery of both DNA and siRNA into different cell lines. We describe the protocol of chip fabrication, apparatus setup, and cell electroporation assay. Typically, the fabrication of the devices takes 1 or 2 days and the continuous electroporation assay takes 1 h.

Key words

Electroporation Microfluidic Gene delivery Laminar flow 



This work was supported by the National Natural Science Foundation of China (61176111 and 61204118).


  1. 1.
    Goodnow CC, Vinuesa CG, Randall KL, Mackay F, Brink R (2010) Control systems and decision making for antibody production. Nat Immunol 11:681–688PubMedCrossRefGoogle Scholar
  2. 2.
    Lin Y, Jen C, Huang M, Wu C, Lin X (2001) Electroporation microchips for continuous gene transfection. Sens Actuator B-Chem 79:137–143CrossRefGoogle Scholar
  3. 3.
    Shin YS, Cho K, Kim JK et al (2004) Electrotransfection of mammalian cells using microchannel-type electroporation chip. Anal Chem 76:7045–7052PubMedCrossRefGoogle Scholar
  4. 4.
    Kim JA, Cho K, Shin YS, Jung N, Chung C, Chang JK (2007) A multi-channel electroporation microchip for gene transfection in mammalian cells. Biosens Bioelectron 22:3273–3277PubMedCrossRefGoogle Scholar
  5. 5.
    Geng T, Zhan Y, Wang H, Witting SR, Cornetta KG, Lu C (2010) Flow-through electroporation based on constant voltage for large-volume transfection of cells. J Control Release 144:91–100PubMedCrossRefGoogle Scholar
  6. 6.
    Wang J, Stine MJ, Lu C (2007) Microfluidic cell electroporation using a mechanical valve. Anal Chem 79:9584–9587PubMedCrossRefGoogle Scholar
  7. 7.
    Zhan Y, Wang J, Bao N, Lu C (2009) Electroporation of cells in microfluidic droplets. Anal Chem 81:2027–2031PubMedCrossRefGoogle Scholar
  8. 8.
    Kim SK, Kim JH, Kim KP, Chung TD (2007) Continuous low-voltage dc electroporation on a microfluidic chip with polyelectrolytic salt bridges. Anal Chem 79:7761–7766PubMedCrossRefGoogle Scholar
  9. 9.
    Zhu T, Luo C, Huang J, Xiong C, Ouyang Q, Fang J (2010) Electroporation based on hydrodynamic focusing of microfluidics with low dc voltage. Biomed Microdevices 12:35–40PubMedCrossRefGoogle Scholar
  10. 10.
    Wei Z, Zhao D, Li X et al (2011) A laminar flow electroporation system for efficient DNA and siRNA delivery. Anal Chem 83:5881–5887PubMedCrossRefGoogle Scholar
  11. 11.
  12. 12.
  13. 13.
  14. 14.
  15. 15.
  16. 16.
    Biswas K, Kal S (2006) Etch characteristics of KOH, TMAH and dual doped TMAH for bulk micromachining of silicon. Microelectron J 37:519–525CrossRefGoogle Scholar
  17. 17.
    Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C (1991) A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods 139:271–279PubMedCrossRefGoogle Scholar
  18. 18.
    Jo B, Van Lerberghe LM, Motsegood KM, Beebe DJ (2000) Three-dimensional micro-channel fabrication in polydimethylsiloxane (PDMS) elastomer. J MEMS 9:76–81CrossRefGoogle Scholar
  19. 19.
    Haubert K, Drier T, Beebe D (2006) PDMS bonding by means of a portable, low-cost corona system. Lab Chip 6:1548–1549PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Zewen Wei
    • 1
  • Zhihong Li
    • 2
  1. 1.National Center for Nanoscience and TechnologyBeijingChina
  2. 2.National Key Laboratory of Science and Technology on Micro/Nano FabricationInstitute of Microelectronics, Peking UniversityBeijingChina

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