Carlo Liberale

Nanoscale chemical mapping using three-dimensional adiabatic compression of surface plasmon polaritons

The fields of plasmonics, Raman spectroscopy and atomic force microscopy have recently undergone considerable development, but independently of one another. By combining these techniques, a range of complementary information could be simultaneously obtained at a single molecule level. Here, we report the design, fabrication and application of a photonic-plasmonic device that is fully compatible with atomic force microscopy and Raman spectroscopy. Our approach relies on the generation and localization of surface plasmon polaritons by means of adiabatic compression through a metallic tapered waveguide to create strongly enhanced Raman excitation in a region just a few nanometres across. The tapered waveguide can also be used as an atomic force microscope tip. Using the device, topographic, chemical and structural information about silicon nanocrystals may be obtained with a spatial resolution of 7 nm.

Nano-patterned SERS substrate: application for protein analysis vs. temperature.

We have illustrated the fabrication of nano-structures as a surface enhanced Raman scattering (SERS) substrate using electro-plating and electron-beam lithography techniques to obtain an array of gold nanograin-aggregate structures of diameter ranging between 80 and 100 nm with interstitial gap of 10-30 nm. The nanostructure based SERS substrate permits us to have better control and reproducibility on generation of plasmon polaritons. The calculation shows the possible detection of myoglobin concentration down to attomole. This SERS substrate is used to investigate the structural changes of different proteins; lysozyme, ribonuclease-B, bovin serum albumin and myoglobin in the temperature range between -65 and 90 degrees C. The in-depth analysis even for small conformational changes is performed using 2D Raman correlation analysis and difference Raman analysis in order to gain straightforward understanding of proteins undergoing thermodynamical perturbation.

Cells preferentially grow on rough substrates

Substrate nanotopography affects cell adhesion and proliferation and is fundamental to the rational design of bio-adhesives, to tissue engineering and to the development of assays for in-vitro screening. Cell behavior on rough substrates is still elusive, and the results presented in the open literature remain controversial. Here, the proliferation of cells on electrochemically etched silicon substrates with different roughness and nearly similar surface energy was studied over three days with confocal and atomic force microscopy. The surface profile of the substrates is a self-affine fractal with a roughness R(a) growing with the etching time from approximately 2 to 100 nm and a fractal dimension D ranging between about 2 (nominally flat surface) and 2.6. For four cell types, the number of adhering cells and their proliferation rates exhibited a maximum on moderately rough (R(a) approximately 10-45 nm) nearly Brownian (D approximately 2.5) substrates. The observed cell behavior was satisfactorily interpreted within the theory of adhesion to randomly rough solids. These findings demonstrated the importance of nanogeometry in cell stable adhesion and growth, suggesting that moderately rough substrates with large fractal dimension could selectively boost cell proliferation.

3D nanostar dimers with a sub-10-nm gap for single-/few-molecule surface-enhanced raman scattering.

Plasmonic nanostar-dimers, decoupled from the substrate, have been fabricated by combining electron-beam lithography and reactive-ion etching techniques. The 3D architecture, the sharp tips of the nanostars and the sub-10 nm gap size promote the formation of giant electric-field in highly localized hot-spots. The single/few molecule detection capability of the 3D nanostar-dimers has been demonstrated by Surface-Enhanced Raman Scattering.

Superhydrophobic Surfaces as Smart Platforms for the Analysis of Diluted Biological Solutions

The aim of this paper is to expound on the rational design, fabrication and development of superhydrophobic surfaces (SHSs) for the manipulation and analysis of diluted biological solutions. SHSs typically feature a periodic array or pattern of micropillars; here, those pillars were modified to incorporate on the head, at the smallest scales, silver nanoparticles aggregates. These metal nanoclusters guarantee superior optical properties and especially SERS (surface enhanced Raman scattering) effects, whereby a molecule, adsorbed on the surface, would reveal an increased spectroscopy signal. On account of their two scale-hybrid nature, these systems are capable of multiple functions which are (i) to concentrate a solution, (ii) to vehicle the analytes of interest to the active areas of the substrate and, therefore, (iii) to measure the analytes with exceptional sensitivity and very low detection limits. Forasmuch, combining different technologies, these devices would augment the performance of conventional SERS substrates and would offer the possibility of revealing a single molecule. In this work, similar SHSs were used to detect Rhodamine molecules in the fairly low atto molar range. The major application of this novel family of devices would be the early detection of tumors or other important pathologies, with incredible advances in medicine.

Integrated microfluidic device for single-cell trapping and spectroscopy

Optofluidic microsystems are key components towards lab-on-a-chip devices for manipulation and analysis of biological specimens. In particular, the integration of optical tweezers (OT) in these devices allows stable sample trapping, while making available mechanical, chemical and spectroscopic analyses.

Lipid Droplets: A New Player in Colorectal Cancer Stem Cells Unveiled by Spectroscopic Imaging

The cancer stem cell (CSC) model is describing tumors as a hierarchical organized system and CSCs are suggested to be responsible for cancer recurrence after therapy. The identification of specific markers of CSCs is therefore of paramount importance. Here, we show that high levels of lipid droplets (LDs) are a distinctive mark of CSCs in colorectal (CR) cancer. This increased lipid content was clearly revealed by label‐free Raman spectroscopy and it directly correlates with well‐accepted CR‐CSC markers as CD133 and Wnt pathway activity. By xenotransplantation experiments, we have finally demonstrated that CR‐CSCs overexpressing LDs retain most tumorigenic potential. A relevant conceptual advance in this work is the demonstration that a cellular organelle, the LD, is a signature of CSCs, in addition to molecular markers. A further functional characterization of LDs could lead soon to design new target therapies against CR‐CSCs. Stem Cells 2015;33:35–44

Water soluble nanoporous nanoparticle for in vivo targeted drug delivery and controlled release in B cells tumor context.

Multitasking nanoparticles are gaining great attention for smart drug delivery systems. The exploration of the nano-scale opens new concrete opportunities for revealing new properties and undiscovered cell-particle interactions. Here we present a biodegradable nanoporous silicon nanoparticle that can be successfully employed for in vivo targeted drug delivery and sustained release. The bare nanoporous nanocarriers can be accurately designed and fabricated with an effective control of porosity, surface chemistry and particle size, up to a few nm. The proposed nanoparticles exhibit several remarkable features including high payload, biodegradability, no toxicity, and multiple loading in water without the need of additional chemical reagents at room temperature. The targeting strategy is based on phage display technology that was successfully used to discover cell surface binding peptide for murine B lymphoma A20 cell line. The peptide used in combination with the nanoporous nanoparticles allows an efficient in vivo targeting, a sustained release and a sensible therapeutic effect.

Far-field spectral characterization of conical emission and filamentation in Kerr media

By use of an imaging spectrometer we map the far-field (theta-lambda) spectra of 200-fs optical pulses that have undergone beam collapse and filamentation in a Kerr medium. By studying the evolution of the spectra with increasing input power and by using a model based on an asymptotic linear superposition of stationary wave modes (rather than the exact instantaneous solution), we are able to trace a consistent model of optical beam collapse highlighting the interplay between conical emission, multiple pulse splitting, and other effects such as spatial chirp.

Molding of Plasmonic Resonances in Metallic Nanostructures: Dependence of the Non-Linear Electric Permittivity on System Size and Temperature

In this paper, we review the principal theoretical models through which the dielectric function of metals can be described. Starting from the Drude assumptions for intraband transitions, we show how this model can be improved by including interband absorption and temperature effect in the damping coefficients. Electronic scattering processes are described and included in the dielectric function, showing their role in determining plasmon lifetime at resonance. Relationships among permittivity, electric conductivity and refractive index are examined. Finally, a temperature dependent permittivity model is presented and is employed to predict temperature and non-linear field intensity dependence on commonly used plasmonic geometries, such as nanospheres.