Improving image contrast in fluorescence microscopy with nanostructured substrates.

ABSTRACT

Metallic and dielectric nanostructures can show sharp contrasted resonances, sensitive to the environment, and high field enhancement in sub-wavelength volumes. For this reason, these structures are commonly used as molecular sensors. Only few works have focused on their application in optical microscopy, in particular in superresolution. In this work we have designed, fabricated and optically tested a nanostructured TiO(2) substrate, fabricated by direct embossing of TiO(2) derived film, as a substrate for fluorescence microscopy. Moreover, using numerical simulations, we have compared the signal to background noise with respect to other metallo-dielectric structures. We show that the TiO(2) structure is a good candidate for reducing the thickness of the fluorescence excitation down to ~100 nm. Therefore, this substrate can be used to obtain Total Internal Reflection (TIRF) axial resolution without a TIRF-Microscopy system.