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Sensors 2020, 20, 1333 3 of 15 Germany), Nexterion Spot buffer from Schott (Mainz, Germany), and sodium phosphate buffer from Medicago (Uppsala, Sweden). Hybridization mixes consisted of Bst 2.0 WarmStart DNA Polymerase, Isothermal Amplification Buffer and MgSO4 all purchased from New England Biolabs (Frankfurt, Germany) diluted in DNase/RNase-free water ordered from Life Technologies (Darmstadt, Germany). All used probes/oligonucleotides were synthesized by Biomers (Ulm, Germany). 2.2. Inkjet Printing Initially, the COC substrates were cleaned in isopropanol combined with an ultra-sonic treatment (Transsonic 420/H from Elma Schmidbauer GmbH, Singen, Germany) for 3 min at 30 ◦C. Subsequently the substrates were flushed with de-ionized (DI) water and thermally dried in an oven (UF 55PLUS from Memmert GmbH and Co. KG, Schwabach, Germany) at 80 ◦C for 1 h. A Dimatix DMP 2831 drop-on-demand inkjet-printer from FUJIFILM Dimatix, Inc. with a 10 pl drop size printhead was used. Images of the used waveforms and the printing parameters can be seen in Figures S1–S3 in the supplementary information (SI). The substrate temperature during the printing process of gold and silver ink was set up to 40 ◦C. Because of the low Au wt.% of the gold ink the Au electrodes were printed multilayered. Nanoparticle inks usually need a sintering step for generating conductive paths. Therefore, a thermal sintering in an oven (UF 55PLUS from Memmert GmbH and Co. KG, Schwabach, Germany) at 130 ◦C for 1 h was performed. For a further improvement of the conductivity of the printed structures a combination of thermal sintering (1 h/130 ◦C) and a subsequent photonic curing (energy: 955 mJ/cm2, one pulse with 270 V and a pulse duration of 1 s, PulseForge® 1200 (PF) from NovaCentrix, Austin, USA) was performed, too. The suitability of photonic curing of nanoparticle inks on temperature-sensitive substrate materials such as polyester, polyvinyl chloride or polyethylene was reported before [20,21]. COC as a hydrophobic thermoplastic causes a poor wetting behavior of the Au and Ag ink on the substrate surface. For improving the wetting behavior of the inks on the COC substrate, a pretreatment of the COC surface was necessary. A plasma treatment is a common process for improving the wettability of polymer substrate materials. Hwang et al. have reported the improvement of the wetting of COC after a treatment with oxygen plasma [22]. Therefore, a low-pressure plasma (O2) was tested. For the low-pressure oxygen plasma treatment a PlasmaPrep2 from Diener electronic GmbH + Co. KG (Ebhausen, Germany) was used. The adjustable parameters are the power output, the oxygen flow rate and the processing time. For the optical and electrical characterization, conductive paths with a length of 1 cm were printed with different widths (see SI Figure S4). The influence of the thickness of the conductive paths on the conductivity was investigated by varying the number of printed layers (3 layers, 6 layers and 9 layers). Additionally, squares were printed for the cross cut test (Figure S4B). Two kinds of inkjet-printed electrode (IPE) arrays were realized. The layout (displayed in SI Figure S5) of 2D IPE arrays was adapted from screen-printed 8X220AT gold electrode arrays (SPE), which were purchased from DropSens (Llanera, Spain). 8 working electrodes (WE) of 2.56 mm diameter, counter (CE), and reference electrodes (RE), each, were printed on a planar COC substrate. Substrate thickness was 1 mm, except for the area where the contact pads were printed. There the thickness was reduced to 0.75 mm to reduce the risk of mechanical abrasion. 2D arrays were used to characterize the properties of inkjet-printed structures for DNA hybridization detection in more detail. A 2.5D array, consisting of 6 WE (1 mm diameter), 1 CE, and 1 RE per chamber, was printed on the bottom of a 0.5 mm deep microfluidic chamber (Figure S6). The traces that connect electrodes and contact pads were printed over the sidewalls of the chambers (45◦ slope, 0.2 mm edge radii). The WE and the CE were printed of Au. The RE and contact pads were printed of Ag atop Au structures that serve as adhesion layer. A printed passivation layer of SU-8 ensured that the area of the WE was well-defined for the functionalization step and that traces did not contribute to the measurements e.g., as background signal. Different numbers of Au layers were printed wet-on-wetPDF Image | Inkjet-Printing Nanoparticle Gold Silver Ink Cyclic Olefin
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