DNA biosensor using fluorescence microscopy and impedance spectroscopy

Two types of DNA biosensors are presented. Both sensing principles are demonstrated using synthetic oligomer single-stranded DNA (ssDNA) with concentrations in the micromolar range. A first sensor type is based on the detection of fluorescently labeled ssDNA to a complementary probe that is bound to a silicon substrate by a disuccinimidyl terephtalate and aminosilane immobilization procedure. An enhanced fluorescent response is obtained using constructive interference effects in a fused silica layer deposited before immobilization onto the silicon substrate. The selectivity of different DNA probes towards complementary and non-complementary DNA targets is tested. A second type of DNA sensor is based on the impedimetric response of a solution of unlabeled 20-mer ssDNA in de-ionized water. Interdigitated microelectrodes that are 5 μm wide and separated by 5 μm gaps are microfabricated on glass substrates and the complex impedance of the system in the 100 Hz–100 MHz frequency range is investigated. The proportionality between the measured solution resistance and ssDNA concentration is demonstrated.