article · 01/01/2009
Microfluidic Patterning of Miniaturized DNA Arrays on Plastic Substrates
Résumé
This paper describes the patterning or DNA arrays on plastic surfaces using an elastomeric, two-dimensional microcapillary system (mu CS). Fluidic structures were realized through hot-embossing lithography using Versaflex CL30. Like elastomers based on poly(dimethylsiloxane), this thermoplastic block copolymer is able to seal a surface in a reversible manner, making it possible to confine DNA probes with a level of control that is unparalleled using standard microspouting techniques, We focus on mu CSs that support arrays comprising up to 2 x 48 spots, each being 45 mu m in diameter. Substrates were fabricated from two hard thermoplastic materials, poly(mechylmethacrylate) and a polycyclic olefin (e.g., Zeonor 1060R), which were both activated with 1-ethyl-3-{[}3-(dimethylamino)propyl]carbodiimide hydrochloride and N-hydroxysuccinimide to mediate covalent attachment of DNA molecules, The approach was exemplified by using 0.25-32 mu M solutions of amino-modified oligonucleotides labeled with either Cy3 or Cy5 fluorescent dye in phosphate-buffered saline, allowing for a direct and sensitive characterization of the printed arrays. Solutions were incubated for durations of 1 to > 48 h at 22, 30, and 40 degrees C to probe the conditions for obtaining uniform spots OF high fluorescence intensity. The length (l) and depth (d) of microfluidic supply channels were both important with respect to depletion as well as evaporation or the solvent. While selective activation or the substrate proved helpful to limit unproductive loss of oligonucleotides along trajectories, incubation of solution in a humid environment was necessary to prevent uncontrolled drying of the liquid, keeping the immobilization process intact over extended periods of time. When combined, these strategies effectively promoted the formation of high-quality DNA arrays, making it possible to arrange multiple probes in parallel with a high degree of uniformity, Moreover, we show that resultant arrays are compatible with standard hybridization protocols, which allowed for reliable discrimination of individual strands when exposed to a specific ssDNA target molecule.
Citer cet article
Geissler, M., Roy, E., Diaz-Quijada, G.-A., Galas, J.-C., & Veres, T. (2009). Microfluidic Patterning of Miniaturized DNA Arrays on Plastic Substrates. ACS Appl. Mater. Interfaces, 1(7). https://doi.org/10.1021/am900285g@article{Geissler2009_117,
author = {Geissler, Matthias and Roy, Emmanuel and Diaz-Quijada, Gerardo A. and Galas, Jean-Christophe and Veres, Teodor},
year = {2009},
month = {1},
title = {Microfluidic Patterning of Miniaturized DNA Arrays on Plastic Substrates},
journal = {ACS Appl. Mater. Interfaces},
publisher = {AMER CHEMICAL SOC},
volume = {1},
number = {7},
address = {1155 16TH ST, NW, WASHINGTON, DC 20036 USA},
abstract = {This paper describes the patterning or DNA arrays on plastic surfaces using an elastomeric, two-dimensional microcapillary system (mu CS). Fluidic structures were realized through hot-embossing lithography using Versaflex CL30. Like elastomers based on poly(dimethylsiloxane), this thermoplastic block copolymer is able to seal a surface in a reversible manner, making it possible to confine DNA probes with a level of control that is unparalleled using standard microspouting techniques, We focus on mu CSs that support arrays comprising up to 2 x 48 spots, each being 45 mu m in diameter. Substrates were fabricated from two hard thermoplastic materials, poly(mechylmethacrylate) and a polycyclic olefin (e.g., Zeonor 1060R), which were both activated with 1-ethyl-3-\{[\}3-(dimethylamino)propyl]carbodiimide hydrochloride and N-hydroxysuccinimide to mediate covalent attachment of DNA molecules, The approach was exemplified by using 0.25-32 mu M solutions of amino-modified oligonucleotides labeled with either Cy3 or Cy5 fluorescent dye in phosphate-buffered saline, allowing for a direct and sensitive characterization of the printed arrays. Solutions were incubated for durations of 1 to > 48 h at 22, 30, and 40 degrees C to probe the conditions for obtaining uniform spots OF high fluorescence intensity. The length (l) and depth (d) of microfluidic supply channels were both important with respect to depletion as well as evaporation or the solvent. While selective activation or the substrate proved helpful to limit unproductive loss of oligonucleotides along trajectories, incubation of solution in a humid environment was necessary to prevent uncontrolled drying of the liquid, keeping the immobilization process intact over extended periods of time. When combined, these strategies effectively promoted the formation of high-quality DNA arrays, making it possible to arrange multiple probes in parallel with a high degree of uniformity, Moreover, we show that resultant arrays are compatible with standard hybridization protocols, which allowed for reliable discrimination of individual strands when exposed to a specific ssDNA target molecule.},
url = {http://www.dx.doi.org/10.1021/am900285g},
doi = {10.1021/am900285g},
issn = {1944-8244},
}TY - JOUR
AU - Geissler, Matthias
AU - Roy, Emmanuel
AU - Diaz-Quijada, Gerardo A.
AU - Galas, Jean-Christophe
AU - Veres, Teodor
PY - 2009
DA - 2009/01/01
TI - Microfluidic Patterning of Miniaturized DNA Arrays on Plastic Substrates
JO - ACS Appl. Mater. Interfaces
VL - 1
IS - 7
PB - AMER CHEMICAL SOC
SN - 1944-8244
AB - This paper describes the patterning or DNA arrays on plastic surfaces using an elastomeric, two-dimensional microcapillary system (mu CS). Fluidic structures were realized through hot-embossing lithography using Versaflex CL30. Like elastomers based on poly(dimethylsiloxane), this thermoplastic block copolymer is able to seal a surface in a reversible manner, making it possible to confine DNA probes with a level of control that is unparalleled using standard microspouting techniques, We focus on mu CSs that support arrays comprising up to 2 x 48 spots, each being 45 mu m in diameter. Substrates were fabricated from two hard thermoplastic materials, poly(mechylmethacrylate) and a polycyclic olefin (e.g., Zeonor 1060R), which were both activated with 1-ethyl-3-{[}3-(dimethylamino)propyl]carbodiimide hydrochloride and N-hydroxysuccinimide to mediate covalent attachment of DNA molecules, The approach was exemplified by using 0.25-32 mu M solutions of amino-modified oligonucleotides labeled with either Cy3 or Cy5 fluorescent dye in phosphate-buffered saline, allowing for a direct and sensitive characterization of the printed arrays. Solutions were incubated for durations of 1 to > 48 h at 22, 30, and 40 degrees C to probe the conditions for obtaining uniform spots OF high fluorescence intensity. The length (l) and depth (d) of microfluidic supply channels were both important with respect to depletion as well as evaporation or the solvent. While selective activation or the substrate proved helpful to limit unproductive loss of oligonucleotides along trajectories, incubation of solution in a humid environment was necessary to prevent uncontrolled drying of the liquid, keeping the immobilization process intact over extended periods of time. When combined, these strategies effectively promoted the formation of high-quality DNA arrays, making it possible to arrange multiple probes in parallel with a high degree of uniformity, Moreover, we show that resultant arrays are compatible with standard hybridization protocols, which allowed for reliable discrimination of individual strands when exposed to a specific ssDNA target molecule.
DO - 10.1021/am900285g
UR - http://www.dx.doi.org/10.1021/am900285g
ER -