Skip to main content

Impedimetric Detection for DNA Hybridization Within Microfluidic Biochips

  • Protocol
Microchip-Based Assay Systems

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 385))

Abstract

A fully integrated biochip for the performance of microfluidic-based DNA bioassays is presented. A microlithographically fabricated circumferential interdigitated electrode array of 1- to 5-µm critical line and space dimensions, with associated large area counterelectrode (1000 × WE) and reference electrode (Ag/AgCl), has been developed as a four-electrode system for the electrochemical detection of DNA hybridization using any of the techniques of amperometry, voltammetry, potentiometry, and impedimetry. This is presented as an alternative to optical detection with an emphasis on label-free impedimetric detection of hybridization. A micro total analysis system (µTAS) is presented, using fluidic channels to connect integrated reaction domains with downstream electrochemical detection. This is accomplished by bonding a patterned poly(dimethylsiloxane) (PDMS) substrate to the biochip or by adhesive bonding of the chip to channels fabricated within glass and plastic microfluidic cards, adding increased functionality to the device.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Guiseppi-Elie, A. (2003) Biochip platform for DNA diagnostics. PharmaTech 87, 1–6.

    Google Scholar 

  2. Landers, J. P. (2003) Molecular diagnostic analysis using electrophorectic microchips. Anal. Chem. 75, 2919–2927.

    Article  CAS  Google Scholar 

  3. Breadmore, M. C., Wolfe, K. A., Arcibal, I. G., et al. (2003) Microchip-based purification of DNA from biological samples. Anal. Chem. 75, 1880–1886.

    Article  CAS  Google Scholar 

  4. Schena, M., ed. (1999) DNA Microarrays: A Practical Approach, Oxford University Press, New York.

    Google Scholar 

  5. Guiseppi-Elie, A. and Lingerfelt, L. (2005) Impedimetric detection of DNA hybridization: towards near patient DNA diagnostics, in Immobilisation of DNA on Chips 1(Wittmann, C., ed.), Springer-Verlag, New York.

    Google Scholar 

  6. Gheorge, M. and Guiseppi-Elie, A. (2003) Electrical frequency dependent characterization of DNA hybridization. Bios. Bioelectron. 19, 95–102.

    Article  Google Scholar 

  7. Hang, T. C. and Guiseppi-Elie, A. (2004) Frequency dependent and surface characterization of DNA immobilization and hybridization. Bios. Bioelectron. 19, 1537–1548.

    Article  CAS  Google Scholar 

  8. Guiseppi-Elie, A., Wallace, G. G., and Matsue, T. (1998) Chemical and biological sensors based on electrically conducting polymers, in Handbook of Conducting Polymers (Skotheim, T. A., Elsenbaumer, R. L., and Reynolds, J. R., eds.), Marcel Dekker, Inc., New York.

    Google Scholar 

  9. Gheorge, M. and Guiseppi-Elie, A. (2000) Low-density arrays of DNA-doped polypyrrole. Proc. Am. Chem. Soc. PMSE 83, 550–551.

    Google Scholar 

  10. Lei, C., Gheorge, M., and Guiseppi-Elie, A. (2000) DNA immobilization and bioelectric detection based on conducting polymers. Proc. Am. Chem. Soc. PMSE 83, 552–553.

    CAS  Google Scholar 

  11. Popovich, N. D., Eckhardt, A. E., Mikulecky, J. C., Napier, M. E., and Thomas, R. S. (2002) Electrochemical sensor for detection of unmodified nucleic acids. Talanta 56, 821–828.

    Article  CAS  Google Scholar 

  12. Sheppard, J. and Guiseppi-Elie, A. (1998) Enzyme sensors based on conductimetric measurement, in Enzyme and Microbial Biosensors: Techniques and Protocols (Mulchandani, A. and Rogers, K. R., eds.), Humana Press, Totowa, NJ.

    Google Scholar 

  13. Kell, D. B. (1987) The principles and potential of electrical admittance spectroscopy: an introduction, in Biosensors: Fundamentals and Applications (Turner, A., Karube, I., and Wilson, G. S., eds.), Oxford University Press, Oxford, pp. 427–468

    Google Scholar 

  14. Lamture, J. B., Beattie, K. L., Burke, B. E., et al. (1994) Direct detection of nucleic acid hybridization on the surface of a charge coupled device. Nucleic Acids Res. 22, 2121–2125.

    Article  CAS  Google Scholar 

  15. Maskos, U. and Southern, E. M. (1992) Oligonucleotide hybridizations on glass supports: a novel linker for oligonucleotide synthesis and hybridization properties of oligonucleotides synthesized in situ. Nucleic Acids Res. 20, 1679–1684.

    Article  CAS  Google Scholar 

  16. Sales, J. A. A., Prado, A. G. S., and Airoldi, C. (2002) The incorporation of propane-1,3-diamine into silylant epoxide group through homogenous and heterogeneous routes. Polyhedron 21, 2647–2651.

    Article  CAS  Google Scholar 

  17. Taylor, S., Smith, S., Windle, B., and Guiseppi-Elie, A. T. (2003) Impact of surface chemistry and blocking strategies on DNA microarrays. Nucleic Acids Res. 31, e87.

    Article  Google Scholar 

  18. Duffy, D. C., McDonald, J. C., Schueller, O. J. A., and Whitesides, G. M. (1998) Rapid prototyping of microfluidic systems in poly(dimethylsiloxane). Anal. Chem. 70, 4974–4984.

    Article  CAS  Google Scholar 

  19. Lorenz, H., Despont, M., Fahrni, N., LaBianca, N., Renaud, P., and Vettiger, P. (1997) SU-8: a low-cost negative resist for MEMS. J. Micromechan. Microeng. 7, 121–124.

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Lingerfelt, L., Karlinsey, J., Landers, J., Guiseppi-Elie, A. (2007). Impedimetric Detection for DNA Hybridization Within Microfluidic Biochips. In: Floriano, P.N. (eds) Microchip-Based Assay Systems. Methods in Molecular Biology™, vol 385. Humana Press. https://doi.org/10.1007/978-1-59745-426-1_8

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-426-1_8

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-588-0

  • Online ISBN: 978-1-59745-426-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics