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IIT – Italian Institute of Technology, Nanophysics, Genoa, Italy

The Italian Institute of Technology (IIT) is a foundation established jointly by the Italian Ministry of Education, Universities and Research and the Ministry of Economy and Finance to promote excellence in basic and applied research and to contribute to the economic development of Italy. The primary goals of the IIT are the creation and dissemination of scientific knowledge as well as the strengthening of Italy's technological competitiveness. To achieve these two goals, the IIT will cooperate with both academic institutions and private organizations, fostering through these partnerships scientific development, technological advances and training in high technology.

http://www.iit.it/en/home.html

The Nanophysics unit, established at the Italian Institute of Technology and abbreviated to "NANOPHYS", aims to design, realize and utilize advanced methodologies and instrumentations within the framework of optical spectroscopy and microscopy, scanning force microscopy and optical nanoscopy.

The unit is oriented to the study and characterization of nanostructured, biological and hybrid materials at the nanoscale, i.e. having at least one of the here spatial dimensions controllable at the nanometric or subnanometric scale.

http://www.iit.it/en/nanophysics.html

  • STED-FLCS: An Advanced Tool to Reveal Spatiotemporal Heterogeneity of Molecular Membrane Dynamics

    Heterogeneous diffusion dynamics of molecules play an important role in many cellular signaling events, such as of lipids in plasma membrane bioactivity. However, these dynamics can often only be visualized by single-molecule and super-resolution optical microscopy techniques. Using fluorescence lifetime correlation spectroscopy (FLCS, an extension of fluorescence correlation spectroscopy, FCS ) on a super-resolution stimulated emission depletion (STED) microscope, we here extend previous observations of nanoscale lipid dynamics in the plasma membrane of living mammalian cells.
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  • Two-Photon Excitation STED Microscopy with Time-Gated Detection

    We report on a novel two-photon excitation stimulated emission depletion (2PE-STED) microscope based on time-gated detection. The time-gated detection allows for the effective silencing of the fluorophores using moderate stimulated emission beam intensity. This opens the possibility of implementing an efficient 2PE-STED microscope with a stimulated emission beam running in a continuous-wave.
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  • Gated STED Microscopy with Time-gated Single-photon Avalanche Diode

    The maximization of the useful (within the time gate) photon flux is then an important aspect to obtain super-resolved STED images. Here we show that by using a fast-gated single-photon avalanche diode (SPAD), i.e. a detector able to rapidly (hundreds picoseconds) switch-on and -off can improve significantly the signal-to-noise ratio (SNR) of the gated STED image. In addition to an enhancement of the image SNR, the use of the fast-gated SPAD reduces also the system complexity. We demonstrate these abilities both on calibration and biological sample.
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  • Abstracts of the 5th European Super-Resolution User-Club Meeting

    The 5th Super-Resolution User Club Meeting was held in collaboration with Professor Kees Jalink and The Netherlands Cancer Institute (NKI) in Amsterdam. Having the meeting at a location where super-resolution microscopy is used on a daily basis makes a big difference, offering participants the chance to use live cells for workshops and see systems working in their true environments. Thanks also to the scientists that supported the meeting by coming and giving talks. As super-resolution continues to grow in importance in research, we recognize the need to come together to network, share information and experiences. Here we present the abstracts of the talks.
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  • Encoding and Decoding Spatio-Temporal Information for Super-Resolution Microscopy

    The challenge of increasing the spatial resolution of an optical microscope beyond the diffraction limit can be reduced to a spectroscopy task by proper manipulation of the molecular states. The nanoscale spatial distribution of the molecules inside the detection volume of a scanning microscope can be encoded within the fluorescence dynamics and decoded by resolving the signal into its dynamics components.
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  • STED Nanoscopy: A Glimpse into the Future

    The well-known saying of "Seeing is believing" became even more apt in biology when stimulated emission depletion (STED) nanoscopy was introduced in 1994 by the Nobel laureate S. Hell and coworkers. This article gives an overview of the various cutting-edge implementations of STED nanoscopy and tries to shine a light into the future: imaging everything faster with unprecedented sensitivity and label-free.
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  • Multi-Images Deconvolution Improves Signal-to-Noise Ratio on Gated Stimulated Emission Depletion Microscopy

    Time-gated detection, namely, only collecting the fluorescence photons after a time-delay from the excitation events, reduces complexity, cost, and illumination intensity of a stimulated emission depletion (STED) microscope. In the gated continuous-wave- (CW-) STED implementation, the spatial resolution improves with increased time-delay, but the signal-to-noise ratio (SNR) reduces.
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  • Nanoscale Protein Diffusion by STED-Based Pair Correlation Analysis

    We describe for the first time the combination between cross-pair correlation function analysis (pair correlation analysis or pCF) and stimulated emission depletion (STED) to obtain diffusion maps at spatial resolution below the optical diffraction limit (super-resolution). Our approach was tested in systems characterized by high and low signal to noise ratio, i.e. Capsid Like Particles (CLPs) bearing several (>100) active fluorescent proteins and monomeric fluorescent proteins transiently expressed in living Chinese Hamster Ovary cells, respectively.
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  • Abstracts of the 4th European Super-Resolution User-Club Meeting

    The 4th Super-Resolution User Club Meeting was held in collaboration with Christian Eggeling and the Weatherall Institute of Molecular Medicine in Oxford, UK. Here we present the abstracts of the talks and interviews with participants.
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  • Alberto Diaspro Comments on Receiving the Emily M. Gray Award 2014

    Alberto Diaspro, Director of the Department of Nanophysics at the Italian Institute of Technology (IIT), has received the Emily M. Gray Award 2014 for dedicating his career to mentoring students and serving as the major force in organizing international biophysics workshops that showcase the talent of young researchers. The Biophysical Society (www.biophysics.org) yearly rewards scientists who particularly stood out for their contributions to education in biophysics. In this brief video he comments on what it was like to receive the Award and on his contributions to education and mentorship in biophysics.
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  • A New Filtering Technique for Removing Anti-Stokes Emission Background in Gated CW-STED Microscopy

    Stimulated emission depletion (STED) microscopy is a prominent approach of super-resolution optical microscopy, which allows cellular imaging with so far unprecedented unlimited spatial resolution. The introduction of time-gated detection in STED microscopy significantly reduces the (instantaneous) intensity required to obtain sub-diffraction spatial resolution.
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  • Single-Wavelength Two-Photon Excitation-stimulated Emission Depletion (SW2PE-STED) Superresolution Imaging

    Two-photon microscopy, multiphoton microcopy and super-resolution imaging. We developed a new class of two-photon excitation–stimulated emission depletion (2PE-STED) optical microscope. In this work, we show the opportunity to perform superresolved fluorescence imaging, exciting and stimulating the emission of a fluorophore by means of a single wavelength.
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  • Abstracts of the 3rd European Super-Resolution User-Club Meeting

    The 3rd meeting of the Leica Super-Resolution User Club was held from June 17th to 19th, 2013 in collaboration with Alberto Diaspro and the Italian Institute of Technology (IIT) in Genoa. Confocal and widefield super-resolution users from ten European countries took three days’ out to deepen their knowledge on super-resolution techniques and applications and make use of an opportunity for full exchange of experiences.
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  • STED Nanoscopy with Time-Gated Detection: Theoretical and Experimental Aspects

    In a stimulated emission depletion (STED) microscope the region in which fluorescence markers can emit spontaneously shrinks with continued STED beam action after a singular excitation event. This fact has been recently used to substantially improve the effective spatial resolution in STED nanoscopy using time-gated detection, pulsed excitation and continuous wave (CW) STED beams.
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  • Abstracts of the 2nd European Super-Resolution User-Club Meeting

    The 2nd meeting of the Leica Super-resolution User club was held from September 25 to 27, 2012 in collaboration with the Science for Life Laboratory at the Karolinska Institute, Stockholm, Sweden. With a mixture of engaging talks by key experts in the field of super-resolution microscopy and stimulating discussion sessions, the meeting proved as popular as last year’s event, attracting a wide range of scientists interested in both confocal and widefield super-resolution and sample preparation techniques.
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  • Developments in Multiphoton Excitation Microscopy

    Basics, history, and applications of multiphoton microscopy. Honouring Goeppert-Mayer’s prediction of simultaneous two-photon absorption by an atom or molecule reported in the 1930s in her PhD dissertation thesis, we can state that multiphoton excitation (MPE) microscopy, more frequently identified with two-photon excitation (2PE) fluorescence microscopy, is a key microscopy method in many areas from medicine to biology, from biophysics to materials science.
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  • Order versus Disorder

    In modern biomaterial design the generation of an environment mimicking some of the extracellular matrix features is envisaged to support molecular cross-talk between cells and scaffolds during tissue formation/remodeling. In bone substitutes chemical biomimesis has been particularly exploited; conversely, the relevance of pre-determined scaffold architecture for regenerated bone outputs is still unclear.
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  • Gated STED Microscopy with CW-STED Lasers

    Among the major steps in the development of Stimulated Emission Depletion (STED) microscopy, the demonstration of the use continuous wave lasers (CW-STED) was certainly contributing the most to a wide dissemination of the method due to the affordability and elegant simplicity of this implementation. Nevertheless, CW-STED was so far not reaching the same spatial resolution as pulsed-lasers STED configurations. A recent investigation on the time-course of the fluorescence emission probability in CW-STED has revealed the benefit of using a gated fluorescence detection (gSTED) to improve further the resolution of C-S
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  • Abstracts of the First European Super-Resolution User-Club Meeting

    The first European Super-resolution User-Club meeting took place from October 27 to 29 in Göttingen, Germany. Prof. Stefan Hell, the inventor of the STED technology, has hosted this first meeting. The user club is aimed at pioneering researchers from the European scientific community, who are early adopters and developers of super-resolution techniques.
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  • Stem Cell Biology in Cancer Research

    The comprehension of stem cell biology and its molecular basis is now acquiring paramount importance in cancer research. The need to look at a single, possibly living, cell makes fluorescence microscopy and confocal microscopy invaluable allies in the study of stem cells.
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  • High Data Output and Automated 3D Correlative Light–Electron Microscopy Method

    Correlative light/electron microscopy (CLEM) allows the simultaneous observation of a given subcellular structure by fluorescence light microscopy (FLM) and electron microscopy. The use of this approach is becoming increasingly frequent in cell biology. In this study, we report on a new high data output CLEM method based on the use of cryosections.
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