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doctoral thesis

Biological samples studied by optical nanospectroscopy

Generosi, Johanna  
2009

The different parts of the electromagnetic spectrum result in diverse effects upon interaction with matter: according to the wavelength, the radiation has energy appropriate for the excitation of a specific physical process. X-rays can be used as a tool to analyze the structure of matter since their wavelength is comparable with the interatomic distances. Infrared light is in the spectral region that excites molecular vibrations and is employed to investigate the chemical composition of a material. Visible radiation can study the optical properties of a sample, such as the fluorescence and the absorbance, and provide a chemical fingerprint when the inelastically scattered light is detected. In this thesis work these light sources are used in diverse experimental approaches to study structured biological specimens, resulting in a detailed chemical and physical characterization at the atomic and molecular scale. Conventional spectroscopy is often not enough sensitive and spatially resolved to detect specific elements or domains in a sample. The need of imaging objects on increasingly finer scales and spatially localize specific molecules, brought to combine infrared, visible and Raman spectroscopy with scanning near-field microscopy giving rise to a powerful nanospectroscopic tool used to perform simultaneous topographical measurements and optical/chemical characterizations with subwavelength resolution, overcoming the diffraction limit of light. Our study combines X-ray diffraction and reflectivity with optical nanospectroscopy to investigate the order and clustering of lipid bilayers, the interaction between solid-supported membranes and embedded alamethicin peptides, the optical and chemical properties of hippocampal neuron cells and the trafficking mechanism of specific neuron receptors.

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