Parallel Multispot Detection of Target Hybridization to Surface-Bound Probe Oligonucleotides of Different Base Mismatch by Surface-Plasmon Field-Enhanced Fluorescence Microscopy†

We introduce surface-plasmon field-enhanced fluorescence microscopy for the on-line determination of hybridization reactions between targets from solution to complementary probe oligonucleotides which are surface-bound to a sensor substrate in a matrix format. This way, the parallel and real-time detection of the association and dissociation kinetics (rate constants) of hybrids of different mismatch situations can be quantitatively analyzed.