Ultrasound delivery of Surface Enhanced InfraRed Absorption active gold-nanoprobes into fibroblast cells: a biological study via Synchrotron-based InfraRed microanalysis at single cell level

Ultrasound (US) induced transient membrane permeabilisation has emerged as a hugely promising tool for the delivery of exogenous vectors through the cytoplasmic membrane, paving the way to the design of novel anticancer strategies by targeting functional nanomaterials to specific biological sites. An essential step towards this end is the detailed recognition of suitably marked nanoparticles in sonoporated cells and the investigation of the potential related biological effects. By taking advantage of Synchrotron Radiation fourier transform infrared micro-spectroscopy (SR-microftiR) in providing highly sensitive analysis at the single cell level, we studied the internalisation of a nanoprobe within fibroblasts (NIH-3T3) promoted by low-intensity US. To this aim we employed 20 nm gold nanoparticles conjugated with the IR marker 4-aminothiophenol. The significant Surface Enhanced Infrared Absorption provided by the nanoprobes, with an absorbance increase up to two orders of magnitude, allowed us to efficiently recognise their inclusion within cells. Notably, the selective and stable SR- microftiR detection from single cells that have internalised the nanoprobe exhibited clear changes in both shape and intensity of the spectral profile, highlighting the occurrence of biological effects. Flow cytometry, immunofluorescence and murine cytokinesis-block micronucleus assays confirmed the presence of slight but significant cytotoxic and genotoxic events associated with the US-nanoprobe combined treatments. our results can provide novel hints towards US and nanomedicine combined strategies for cell spectral imaging as well as drug delivery-based therapies

In situ characterization of vertically oriented carbon nanofibers for three-dimensional nano-electro-mechanical device applications

We have performed mechanical and electrical characterization of individual as-grown, vertically oriented carbon nanofibers (CNFs) using in situ techniques, where such high-aspect-ratio, nanoscale structures are of interest for three-dimensional (3D) electronics, in particular 3D nano-electro-mechanical-systems (NEMS). Nanoindentation and uniaxial compression tests conducted in an in situ nanomechanical instrument, SEMentor, suggest that the CNFs undergo severe bending prior to fracture, which always occurs close to the bottom rather than at the substrate–tube interface, suggesting that the CNFs are well adhered to the substrate. This is also consistent with bending tests on individual tubes which indicated that bending angles as large as ~70° could be accommodated elastically. In situ electrical transport measurements revealed that the CNFs grown on refractory metallic nitride buffer layers were conducting via the sidewalls, whereas those synthesized directly on Si were electrically unsuitable for low-voltage dc NEMS applications. Electrostatic actuation was also demonstrated with a nanoprobe in close proximity to a single CNF and suggests that such structures are attractive for nonvolatile memory applications. Since the magnitude of the actuation voltage is intimately dictated by the physical characteristics of the CNFs, such as diameter and length, we also addressed the ability to tune these parameters, to some extent, by adjusting the plasma-enhanced chemical vapor deposition growth parameters with this bottom-up synthesis approach

Optical Polarization M\"obius Strips and Points of Purely Transverse Spin Density

Tightly focused light beams can exhibit electric fields spinning around any axis including the one transverse to the beams' propagation direction. At certain focal positions, the corresponding local polarization ellipse can degenerate into a perfect circle, representing a point of circular polarization, or C-point. We consider the most fundamental case of a linearly polarized Gaussian beam, where - upon tight focusing - those C-points created by transversely spinning fields can form the center of 3D optical polarization topologies when choosing the plane of observation appropriately. Due to the high symmetry of the focal field, these polarization topologies exhibit non trivial structures similar to M\"obius strips. We use a direct physical measure to find C-points with an arbitrarily oriented spinning axis of the electric field and experimentally investigate the fully three-dimensional polarization topologies surrounding these C-points by exploiting an amplitude and phase reconstruction technique.Comment: 5 pages, 3 figures; additional supplementary materials with 4 pages, 3 figure

Mechanical modulation of single-electron tunneling through molecular-assembled metallic nanoparticles

We present a microscopic study of single-electron tunneling in nanomechanical double-barrier tunneling junctions formed using a vibrating scanning nanoprobe and a metallic nanoparticle connected to a metallic substrate through a molecular bridge. We analyze the motion of single electrons on and off the nanoparticle through the tunneling current, the displacement current and the charging-induced electrostatic force on the vibrating nanoprobe. We demonstrate the mechanical single-electron turnstile effect by applying the theory to a gold nanoparticle connected to the gold substrate through alkane dithiol molecular bridge and probed by a vibrating platinum tip.Comment: Accepted by Phys. Rev.

Characterization and scanning probe spectroscopy of radiation-pressure induced mechanical oscillation of a microcavity

Microcavities can enter a regime where radiation pressure causes oscillation of mechanical cavity eigenmodes. We present a detailed experimental and theoretical understanding of this effect, and report direct scanning probe spectroscopy of the micro-mechanical modes

Evaluating a Novel Photochemical Tool for Labeling and Tracking Live, Endogenous Calcium-Permeable AMPARs

The purpose of this research is to advance development of a photochemical tool designed to probe the role of ionotropic glutamate receptor signaling in neurodegenerative processes, and to delve more deeply into the biological processes underlying the role of these receptors in signaling and memory formation. This ligand-targeted nanoprobe was designed and developed in our lab to label endogenous calcium-permeable AMPARs (CP-AMPARs) in live cells with minimal disruption to native receptor activity. Nanoprobe is designed to use naphthyl acetyl spermine (NASPM) as a photocleavable ligand to target and covalently label native CP-AMPARs with a non-perturbing, fluorescent marker that then allows observation of these receptors using standard epifluorescence microscopy. My contribution to this work, outlined in the aims below, is the characterization of nanoprobe using electrophysiology and fluorescent imaging to evaluate its effectiveness as an endogenous CP-AMPAR label on live neurons. Aim 1: To use whole cell patch clamp electrophysiology to test the labeling of CP-AMPARs with nanoprobe by recording changes in glutamate-evoked current through heterologously expressed GluA1-L497Y homomultimers during, pre- and post- nanoprobe labeling. Aim 2: To use fluorescent imaging to evaluate nanoprobe labeling of glutamate receptors endogenously expressed in hippocampal neurons by co-labeling nanoprobe-treated neurons with traditional antibodies to AMPAR and synaptic targets. Aim 3: To use nanoprobe to detect endogenously expressed CP-AMPARs on live neurons during the course of neuron development. Live neuronal cultures will be imaged before and after labeling with nanoprobe in young dissociated cultures (DIV 1-2) and in maturing cultures (DIV 14-17). Conclusions: Whole cell patch clamp electrophysiology results provide evidence that nanoprobe will label CP-AMPARs in a minimally-perturbing fashion that allows the receptors to resume normal activity after photolytic-release of ligand as designed. Fixed cell imaging of CP-AMPAR nanoprobe labeling was largely ineffective, and live cell imaging was not conclusive, but provided supporting evidence that nanoprobe targets and labels NASPM-sensitive endogenous glutamate receptors on live dissociated hippocampal neuron

Sequential Adaptive Detection for In-Situ Transmission Electron Microscopy (TEM)

We develop new efficient online algorithms for detecting transient sparse signals in TEM video sequences, by adopting the recently developed framework for sequential detection jointly with online convex optimization [1]. We cast the problem as detecting an unknown sparse mean shift of Gaussian observations, and develop adaptive CUSUM and adaptive SSRS procedures, which are based on likelihood ratio statistics with post-change mean vector being online maximum likelihood estimators with $\ell_1$. We demonstrate the meritorious performance of our algorithms for TEM imaging using real data

Label-Free Detection of DNA Hybridization with A Compact LSPR-based Fiber-Optic Sensor

A miniaturized, robust, localized surface plasmon resonance (LSPR)-coupled fiber-optic (FO) nanoprobe providing an integrated and portable solution for detection of DNA hybridization and measurement of DNA concentrations has been demonstrated. The FO nanoprobe was created by constructing arrays of metallic nanostructures on the end facets of optical fibers utilizing nanofabrication technologies, including electron beam lithography and lift-off processes. The LSPR-FO nanoprobe device offers real-time, label-free, and low-sample-volume quantification of single-strand DNA in water with high sensitivity and selectivity, achieving a limit of detection around 10 fM. These results demonstrate the feasibility of the LSPR-FO nanoprobe device as a compact and low-cost biosensor for detection of short-strand DNA