"People really interact during the sessions – this is really important and helps to create a nice atmosphere. I look forward to participating in the 3rd meeting", said Giuseppe Vicidomini, one of the speakers at this year's meeting, who specializes in gated confocal super-resolution techniques at the IIT, Genoa, Italy.
"The meeting enables users of Leica Microsystems ground-breaking super-resolution systems to meet and share experiences – we are happy to provide a platform for our customers to do that, and to continue to support the work they do in research science" says Baba Awopetu, European Marketing Director, Leica Microsystems.
Alberto Diaspro, Italian Institute of Technology, Genoa, Italy
It is well established that for fluoresence, the most popular imaging mode in optical microscopy, the diffraction barrier no longer provides an unsurpassable limitation for resolution and localization accuracy. Furthermore, the terms "super-resolution" and the earlier coined "optical nanoscopy", have been implemented in real far field optical microscopes, and are now available for everyone to use without extreme complexity. Since the new actor for getting a resolution improvement is the fluorescent molecule being used and its photophysics is sometimes sample-related, at the Istituto Italiano di Tecnologia (IIT) we decided to attack the problem from different sides.
For this reason, we implemented and adapted a number of different optical fluorescence set-ups towards super-resolution. In considering super-resolution approaches, we also considered their implementation for imaging thick (>50 um)
biological specimens. Within the framework of stochastic read-out methods based on single molecule localization, we implemented individual molecule localization (IML) within a selective plane illumination microscopy (SPIM) architecture in order to address 3D super-resolution imaging.
An application related to mammalian cellular spheroids will be presented. Recently, the very same system has been endowed of the possibility of performing IML-SPIM using two-photon photo-activation. Moreover, an optical solution will be presented to overcome the SPIM request of inserting the sample in a cylindrical sample holder. On the side of the targeted read-out methods like STED, we introduced two-photon excitation including the possibility of using a single wavelength (SW) both for two-photon excitation and STED depletion by implementing a SW-2PE-STED microscope. Since STED provides immediate access to the resolution improvement without requiring computational tools, we expanded the method to direct writing lithography and multimodal super resolution microscopy. Direct writing lithography at super resolution will be outlined demonstrating the possibility of exploiting the STED-like mechanism. On the other side, in terms of multimodal super resolution microscopy, we will show the coupling of STED and Atomic Force Microscopy. This allows us to more precisely and specifically address