The purpose of cell lysis is to obtain intracellular substances such as DNA and proteins for analysis. Commonly used methods include chemical (chemical solution decomposition) and physical (electricity or mechanical force). This study proposes an integrated system using an electroporation and microfluidic device made by micro-photolithography to lyse a single cell and stretch its DNA. The PDMS, as the manufacturing material of the microfluidic device, consists of 2 parts: the cell lytic zone, in which the immobilized cells trapped within the dense microstructure are lysed at a single-cell level, and the DNA stretching and recovery zone. This study showed that in a hypotonic environment (75 mM glucose solution), when electric field conditions were 100 Vpp and 1 kHz, the target cell was lysed and its DNA was released into the solution. When injected with proteinase K, the DNA flowed along the rectangular microstructure and was stretched to a length exceeding 840 μm.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Kotnik, T., Miklavcic, D. & Slivnik, T. Time course of transmembrance voltage induced by time-varying electric fields-a method for theoretical analysis and its application. Bioelectrochem. 45, 3–16 (1998).
Lu, H., Schmidt, M.A. & Jensen, K.F. A microfluidic electroporation device for cell lysis. Lab Chip 5, 23–29 (2005).
Shin, Y.S. et al. Electrotransfection of mammalian cells using microchannel-type electroporation chip. Anal. Chem. 76, 7045–7052 (2004).
Wang, H.Y. & Lu, C. Electroporation of mammalian cells in a microfluidic channel with geometric variation. Anal. Chem. 78, 5158–5164 (2006).
Lee, D.W. & Cho, Y.H. A continuous electrical cell lysis device using a low dc voltage for a cell transport and rupture. Sens. Actu. B 124, 84–89 (2007).
Lin, Y.H. & Lee, G.B. An optically induced cell lysis device using dielectrophoresis. Appl. Phys. Lett. 94, 033901 (2009).
Chung, Y.C. et al. Microfluidic chip for high efficiency DNA extraction. Lab Chip 4, 141–147 (2004).
Liu, R.H., Yang, J., Lenigk, R., Bonanno, J. & Grodzinski, P. Self-contained, full integrated biochip for sample preparation, polymerase chain reaction amplification, and DNA microarray detection. Anal. Chem. 76, 1824–1831 (2004).
Lee, C.Y., Lee, G.B., Lin, J.L., Huang, F.C. & Liao, C.S. Integrated microfluidic systems for cell lysis, mixing/pumping and DNA amplification. J. Micromech. Microeng. 15, 1215–1233 (2005).
Teissié, J. & Rols, M.P. An experimental evaluation of the critical potential difference inducing cell membrane electropermeabilization. Biophys. J. 65, 409–413 (1993).
Hung, M.-S., Kurosawa, O., Kabata, H. & Washizu, M. Stretching DNA fibers out of a chromosome in solution using electroosmotic flow. J. Chin. Soc. Mech. Eng. 30, 289–295 (2009).
Hung, M.-S. & Chen, P.-C. Extending DNA from a single cell using integrated system of electro-osmosis and AFM. J. Med. Biol. Eng. 30, 29–34 (2010).
Lee, S.W. & Tai, Y.C. A micro cell lysis device. Sens. Actu. A 73, 74–79 (1999).
Neumann, E., Kakorin, S., Tsoneva, I., Nikolova, B. & Tomov, T. Calcium-mediated DNA adsorption to yeast cells and kinetics of cell transformation by electroporation. Biophys. J. 71, 868–877 (1996).
Hu, Z., Kwon, G.H., Kim, C.-B., Kim, D. & Lee, S.- H. Integration of movable structures in PDMS microfluidic channels. BioChip J. 4, 117–122 (2010).
About this article
Cite this article
Hung, MS., Chang, YT. Single cell lysis and DNA extending using electroporation microfluidic device. BioChip J 6, 84–90 (2012). https://doi.org/10.1007/s13206-012-6111-x
- Cell lysis
- DNA extending
- Microfluidic device