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"Super resolution optical fluorescence microscopy"

Relatore: Alberto Diaspro- Università di Genova e Dipartimento di Nanofisica, IIT, Genova
Note: Seminario di Dipartimento

Aula Newton
08 Maggio 2013 ore 16.30

Super resolution optical fluorescence microscopy Alberto Diaspro It is well known and established that, for the most popular imaging mode in optical microscopy, i.e. fluorescence, the diffraction barrier does no longer provide an unsurpassable limitation for resolution and localization accuracy. Furthermore, the terms “super resolution” and "optical nanoscopy", coined earlier, have been implemented in real far field optical microscopes, nowadays available for everyone to use without extreme complexity. Here, we will discuss targeted and stochastic readout methods using both single and multiphoton excitation, in terms of resolution and localization precision accuracy. Individual molecule localization (IML) implemented within selective plane illumination microscopy (SPIM) will be addressed towards 3D super resolution imaging in thick biological samples. STED two-photon excitation microscopy will be discussed reporting about the possibility of using a single wavelength (SW) both for two-photon excitation and STED depletion by implementing a SW-2PE-STED microscope. Particular attention will be given to the intensity issues related to the depletion process. Direct writing lithography at super resolution will be discussed along with the characterization of new fluorescent reporter super-resolution oriented. A further topic will be related to the coupling of STED and Atomic Force Microscopy. So far, a variety of architectures will be outlined in regard to specific applications demanding for nanoscale investigations.