Abstract

Near-field Scanning Optical Microscopy (NSOM) in liquid environment is expected to allow time resolved morphological mappings on cellular surfaces on the nanoscale level. Near-field Optical Analysis (NOA) via NSOM exploits the energy transfer from the tip of an optical element (tip diameter ≥20 nm), oscillating within the range of the characteristic length of the energy transfer (∼10 nm) in the near-field of the surface to be analysed. In NOA, a molecular assembly is monitored by visible light with a resolution far below the wavelength of visible light. Actually, NOA is successfully applied in mapping local optical contrasts, for instance in photonic crystals with dielectric periodicities on the nanoscale. NSOM could in principle be performed in two different modes: tapping mode, with tip-oscillations perpendicular, or shear force mode, with tip-oscillations parallel to the substrate. Both basic modes have specific advantages and disadvantages. In biological systems (e.g. in cell cultures), where scanning in liquids is prevalent, elongated optical elements non-invasively operated in the shear force modus could have some specific advantages when compared to contact modus systems. While tapping mode NSOM provides satisfactory nanoscale images even on solid surfaces covered with millimetres of liquids, the performance of shear force mode NSOM is presently largely confined to operations on dry samples. This is due to the inability of conventional shear force mode NSOM systems to provide sharp topographic images of sample surfaces substantially covered with liquids. By equipping a conventional NSOM system with hydrophobic optical elements, shear force mode based topographic images could be obtained on biological samples in dry as well as in aqueous environment, and with resolutions on the nanoscale level.

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