Neuronal cell growth on polymeric scaffolds studied by CARS microscopy

Abstract

Higher-order nonlinearity of light-matter interactions, such as second and third harmonic generation (SHG & THG) and Coherent anti-Stokes Raman Scattering (CARS) can be used for improving spatial resolution in microscopy as a consequence of the confinement of the nonlinear polarization to the high-intensity region of the focal volume. However, the resolution is limited to similar to 300 nm, not sufficient to resolve macromolecules or nanostructures of interest in the bio-, life- and nano-sciences. In the strive to push the resolution beyond the diffraction limit, allowing for nanoscale imaging, we have equipped a nonlinear optical microscope with a scanning-probe setup operated in tapping-mode feedback. A tapered, gold-coated, open-aperture tip with an aperture diameter of similar to 150 nm is scanned over the sample, probing the nonlinear nearfield generated by free-beam excitation. First nonlinear coherent Raman nearfield images of biological macromolecules and metallic nanostructures are shown. Limitations and future challenges with nonlinear nearfield microscopy are discussed.