Enhancing DNA hybridization kinetics through constriction-based dielectrophoresis.

The enhancement of signal sensitivity through the scaling-down of sensors presents mass transport limitations that can arrest the sensitivity gains obtained as a result of miniaturization. To alleviate these limitations, we study the application of constriction-based dielectrophoresis methods to enhance transport through pre-concentration of target DNA in the vicinity of the diffusion layer of the sensor, on which capture probe DNA molecules were immobilized. We demonstrate that constriction-based DEP pre-concentration was not impeded by scaling-down of the sensor, as long as the sensor electrode was composed of nanostructured edges and was coupled to an equally scaled down insulating constriction within a microfluidic channel to enhance the focusing effects of the constrictions and edges. Furthermore, as a result of the high focusing fields, pre-concentration of single-stranded target DNA occurred in the vicinity of the sensor pad within the relatively high ionic strength buffers required for DNA hybridization, with minimal degradation of capture probe molecules. Finally, constriction-based DEP resulted in an almost immediate pre-concentration of target DNA in the vicinity of the sensor electrode diffusion layer, resulting in a ten-fold enhancement of the DNA hybridization kinetics at target concentration values down to the sensitivity limit of 10 pM for the sensor platform.

[1]  A. Singh,et al.  Dielectrophoresis in microchips containing arrays of insulating posts: theoretical and experimental results. , 2003, Analytical chemistry.

[2]  Mikael Käll,et al.  Plasmonic sensing characteristics of single nanometric holes. , 2005, Nano letters.

[3]  H. Yowanto,et al.  Electronic detection of nucleic acids: a versatile platform for molecular diagnostics. , 2001, The Journal of molecular diagnostics : JMD.

[4]  R. Colton,et al.  The BARC biosensor applied to the detection of biological warfare agents. , 2000, Biosensors & bioelectronics.

[5]  V Chu,et al.  Electric-field assisted immobilization and hybridization of DNA oligomers on thin-film microchips , 2005, Nanotechnology.

[6]  Boris Mizaikoff,et al.  Model-based optimal design of polymer-coated chemical sensors. , 2003, Analytical chemistry.

[7]  M. Heller,et al.  Active microelectronic chip devices which utilize controlled electrophoretic fields for multiplex DNA hybridization and other genomic applications , 2000, Electrophoresis.

[8]  Robin H. Liu,et al.  Plastic biochannel hybridization devices: a new concept for microfluidic DNA arrays. , 2002, Analytical biochemistry.

[9]  C Bamdad,et al.  A DNA self-assembled monolayer for the specific attachment of unmodified double- or single-stranded DNA. , 1998, Biophysical journal.

[10]  F. Zenhausern,et al.  Electrodeless dielectrophoresis for micro total analysis systems , 2003, IEEE Engineering in Medicine and Biology Magazine.

[11]  H. Yowanto,et al.  Electronic detection of single-base mismatches in DNA with ferrocene-modified probes. , 2001, Journal of the American Chemical Society.

[12]  P. Sheehan,et al.  Detection limits for nanoscale biosensors. , 2005, Nano letters.

[13]  M. Washizu Bio-nanotechnology of DNA based on electrostatic manipulation , 2004 .

[14]  J. Bishop,et al.  Effects of fill fraction on the capture efficiency of nanoscale molecular transducers , 2006, Nanotechnology.

[15]  Robin H. Liu,et al.  Hybridization enhancement using cavitation microstreaming. , 2003, Analytical chemistry.

[16]  Alexandre G. Brolo,et al.  Nanohole-Enhanced Raman Scattering , 2004 .

[17]  R. Pethig,et al.  Applications of dielectrophoresis in biotechnology. , 1997, Trends in biotechnology.

[18]  C. Mirkin,et al.  Nanoparticles with Raman spectroscopic fingerprints for DNA and RNA detection. , 2002, Science.

[19]  C. Chou,et al.  A simple polysilsesquioxane sealing of nanofluidic channels below 10 nm at room temperature. , 2007, Lab on a chip.

[20]  Muhammad A. Alam,et al.  Performance limits of nanobiosensors , 2006 .

[21]  Chia-Fu Chou,et al.  Electrodeless dielectrophoresis of single- and double-stranded DNA. , 2002, Biophysical journal.

[22]  C. Chou,et al.  Two-potential electrochemical probe for study of DNA immobilization. , 2005, Langmuir : the ACS journal of surfaces and colloids.