lifesciences
Innopsys

Automated and Multiplexed Soft Lithography for the Production of Low-Density DNA Microarrays

Abstract


Microarrays are established research tools for genotyping, expression profiling, or molecular diagnostics in which DNA molecules are precisely addressed to the surface of a solid support. This study assesses the fabrication of low-density oligonucleotide arrays using an automated microcontact printing device, the InnoStamp 40®. This automate allows a multiplexed deposition of oligoprobes on a functionalized surface by the use of a MacroStampTM bearing 64 individual pillars each mounted with 50 circular micropatterns (spots) of 160 μm diameter at 320 μm pitch. Reliability and reuse of the MacroStampTM were shown to be fast and robust by a simple washing step in 96% ethanol. The low-density microarrays printed on either epoxysilane or dendrimer-functionalized slides (DendriSlides) showed excellent hybridization response with complementary sequences at unusual low probe and target concentrations, since the actual probe density immobilized by this technology was at least 10-fold lower than with the conventional mechanical spotting. In addition, we found a comparable hybridization response in terms of fluorescence intensity between spotted and printed oligoarrays with a 1 nM complementary target by using a 50-fold lower probe concentration to produce the oligoarrays by the microcontact printing method. Taken together, our results lend support to the potential development of this multiplexed microcontact printing technology employing soft lithography as an alternative, cost-competitive tool for fabrication of low-density DNA microarrays.

Authors

Fredonnet J1, Foncy J1,2, Cau JC3, Séverac C1, François1,2,4, Trévisiol1,5,6

  • 1. ITAV, Université de Toulouse, CNRS, UPS, Toulouse 31000, France
  • 2. LISBP, Université de Toulouse, INSA, UPS, INP, LISBP, 135 Avenue de Rangueil, Toulouse F-31077, France
  • 3. Innopsys SAS, 3, allée de Vignes, Carbonne 31390, France
  • 4. Dendris SAS, 335 Rue de Chêne Vert, Labège 31670, France
  • 5. CNRS, LAAS, 7 Avenue du Colonel Roche, F-31400 Toulouse, France
  • 6. LAAS, Univ de Toulouse, F-31400 Toulouse, France

 

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