arXiv subjects

Paola Borri

Publications and source records attributed to Paola Borri.

17 recordsLinked to original sources

arXiv2026

Sub-nanometer 3D morphometric precision of polarisation-resolved wide-field optical extinction microscopy determines the roundness of individual gold nanospheres

Quantitative polarisation-resolved optical extinction microscopy of individual plasmonic nanoparticles has recently been introduced as a powerful tool to characterise the nanoparticle's morphology with a precision comparable to electron microscopy, while using a simple optical microscope [Nanoscale 12, 16215 (2020)]. Here we provide a step change by adding measurements for radial polarisation in the condenser back focal plane, probing plasmonic resonances polarised in axial direction. The combined linear and radial polarisation measurements provide a significantly enhanced precision of the retrieved 3D morphology, as we show on defect-free ultra-uniform gold nanospheres of 30 nm nominal diameter characterised by transmission electron microscopy. The measured cross-sections are modelled for an ellipsoidal particle, determining the three semi-axes and rotation angles by fitting the measurements. The material permittivity and surface damping providing the best fit are found. The particle aspect ratio is determined with a precision better than 5%, and the size with an impressive precision of 0.1 nm. Notably, corrections to the Rayleigh-Gans ellipsoid model due to retardation are significant even though the particle diameters are more than an order of magnitude smaller than the wavelength in the medium. Taking them into account improves the shape accuracy.

physics.optics↗

arXiv2025

Understanding the mechanisms of supported lipid membrane reshaping into tubular networks using quantitative DIC microscopy

Biological membranes are known to form various structural motifs, from lipid bilayers to tubular filaments and networks facilitating e.g. adhesion and cell-cell communication. To understand the biophysical processes underpinning lipid-lipid interactions in these systems, synthetic membrane models are crucial. Here, we demonstrate the formation of tubular networks from supported lipid membranes of controlled lipid composition on glass. We quantify tube radii using quantitative differential interference contrast (qDIC) and propose a biophysical mechanism for the formation of these structures, regulated by surface tension and lipid exchange with connected supported membranes. Two lipid types are investigated, namely DOPC and DC15PC, exhibiting a liquid disordered and a solid ordered phase at room temperature, respectively. Tube formation is studied versus temperature, revealing bilamellar layers retracting and folding into tubes upon DC15PC lipids transitioning from liquid to solid phase, which is explained by lipid transfer from bilamellar to unilamellar layers. This study introduces a novel model system for bilayer tubes, allowing to elucidate the biophysics of lipid-lipid interactions governing lipid membrane reshaping into tubular structures, important for our understanding of biological membrane filaments.

cond-mat.soft↗

arXiv2025

Genetically encoding stimulated Raman-scattering probes for cell imaging using infrared fluorescent proteins

Stimulated Raman scattering (SRS) microscopy offers great potential to surpass fluorescent-based approaches, owing to the sharp linewidth of Raman vibrations amenable to super-multiplex cell imaging, but currently lacks one crucial component: genetically encodable tags equivalent to fluorescent proteins. Here, we show that infrared fluorescent proteins (IRFPs) can be used as genetically encoded SRS probes and benefit from the electronic pre-resonant SRS enhancement effect with near-infrared exciting pulses, comparable to synthetic dyes reported in the literature. SRS imaging of the nucleus in mammalian cells is demonstrated where a histone protein is fused to an IRFP. This work opens the route towards Raman-based cell imaging using genetically encoded probes, motivating efforts in solving the challenges of photostability and creating a vibrational palette.

physics.chem-ph↗

arXiv2022

Differential Dynamic Microscopy can be applied to Differential Interference Contrast images despite shadowing effects

During In Vitro Fertilisation (IVF), combining time-lapse contrast microscopy with suitable image processing techniques could facilitate non-invasive classification of oocyte health, driving oocyte selection to maximise success rates. One image processing technique, Differential Dynamic Microscopy (DDM), has been used in a variety of microscopy settings including bright field, dark field and fluorescence. However, in some applications, effects stemming from the choice of microscopy may invalidate underlying assumptions of DDM. Here, we study the DDM analysis of differential interference contrast (DIC) microscopy movies. DIC exhibits a characteristic shadowing effect that gives the illusion of a 3D appearance, which we show causes deformation of the output DDM matrix. We present a mathematical description of this deformation, and conclude that, when studying isotropic motion, no account of the DIC shadow needs to be considered during DDM analysis. We validate our conclusions with simulated particle data, and with DIC images of colloidal dispersions. Although this conclusion does not generally extend to anisotropic motion, we demonstrate that for directed advection and diffusion behaviour, parameter fitting invariance still holds. These results validate the current practice of applying DDM to DIC, and are a foundation for further exploration of DDM in other phase-contrast image datasets.

cond-mat.stat-mech↗

arXiv2022

Correlative extinction and single fluorophore bleaching microscopy for ligand quantification on gold nanoparticles

Nanoparticles (NPs) are promising therapeutic delivery agents, yet it is increasingly apparent that the number and manner of presentation of cell binding ligands on the NP can affect the eventual fate of the therapeutic. Whenever NPs are conjugated with biomolecules, a heterogenous population of decorated NPs will be produced and the details of the subpopulations of particle-ligand structures needs to be characterised for a reliable interpretation of NP-based data. We report an optical microscopy method to quantitatively evaluate the conjugation on a single particle basis in samples consisting of gold NPs (GNPs) decorated with human holo-transferrin fluorescently labelled with Alexa647 (Tf). We employed widefield fluorescence and extinction microscopy on NP-ligand constructs sparesly deposited onto a glass surface, alongside a correlative analysis which spatially co-localises diffraction-limited sources of fluorescence with the optical extinction by individual GNPs. A photobleaching step analysis of the fluorescence emission was employed to estimate the number of fluorophores contributing to the detected emission rate. The method quantifies the number of fluorescent biomolecules attached per GNP, the numbers of unconjugated GNPs and unbound Tf present within the mixed population, and the size and intraparticle clustering propensity of conjugated GNPs. We found a high variability in the number of Tf ligands per GNP within the GNP population, when analysed at the single-particle level, unraveling a non-trivial statistical distribution not accessible in ensemble averaged approaches

physics.optics↗

arXiv2022

Correlative light electron microscopy using small gold nanoparticles as single probes

Correlative light electron microscopy (CLEM) requires the availability of robust probes which are visible both in light and electron microscopy. Here we demonstrate a CLEM approach using small gold nanoparticles as a single probe. Individual gold nanoparticles bound to the epidermal growth factor protein were located with nanometric precision background-free in human cancer cells by light microscopy using resonant four-wave-mixing (FWM), and were correlatively mapped with high accuracy to the corresponding transmission electron microscopy images. We used nanoparticles of 10 nm and 5 nm radius, and show a correlation accuracy below 60 nm over an area larger than 10 um size, without the need for additional fiducial markers. Correlation accuracy was improved to below 40 nm by reducing systematic errors, while the localisation precision is below 10 nm. Polarisation-resolved FWM correlates with nanoparticle shapes, promising for multiplexing by shape recognition in future applications. Owing to the photostability of gold nanoparticles and the applicability of FWM microscopy to living cells, FWM-CLEM opens up a powerful alternative to fluorescence-based methods.

physics.optics↗

arXiv2022

Quantitatively linking morphology and optical response of individual silver nanohedra

The optical response of metal nanoparticles is governed by plasmonic resonances, which are dictated by the particle morphology. A thorough understanding of the link between morphology and optical response requires quantitatively measuring optical and structural properties of the same particle. Here we present such a study, correlating electron tomography and optical micro-spectroscopy. The optical measurements determine the scattering and absorption cross-section spectra in absolute units, and electron tomography determines the 3D morphology. Numerical simulations of the spectra for the individual particle geometry, and the specific optical setup used, allow for a quantitative comparison including the cross-section magnitude. Silver nanoparticles produced by photochemically driven colloidal synthesis, including decahedra, tetrahedra and bi-tetrahedra are investigated. A mismatch of measured and simulated spectra is found when assuming pure silver particles, which is resolved by the presence of a few atomic layers of tarnish on the surface, not evident in electron tomography. The presented method tightens the link between particle morphology and optical response, supporting the predictive design of plasmonic nanomaterials.

physics.optics↗

arXiv2022

uFLIM -- Unsupervised analysis of FLIM-FRET microscopy data

Despite their widespread use in cell biology, fluorescence lifetime imaging microscopy (FLIM) data-sets are challenging to analyse, because each spatial position can contain a superposition of multiple fluorescent components. Here, we present a data analysis method employing all information in the available photon budget, as well as being fast. The method, called uFLIM, determines spatial distributions and temporal dynamics of multiple fluorescent components with no prior knowledge. It goes significantly beyond current approaches which either assume the functional dependence of the dynamics, e.g. an exponential decay, or require dynamics to be known, or calibrated. Its efficient non-negative matrix factorization algorithm allows for real-time data processing. We validate in silico that uFLIM is capable to disentangle the spatial distribution and spectral properties of five fluorescing probes, from only two excitation and detection channels and a photon budget of 100 detected photons per pixel. By adapting the method to data exhibiting Forster resonant energy transfer (FRET), we retrieve the spatial and transfer rate distribution of the bound species, without constrains on donor and acceptor dynamics.

physics.bio-ph↗

arXiv2021

Sizing individual dielectric nanoparticles with quantitative differential interference contrast microscopy

We report a method to measure the size of single dielectric nanoparticles with high accuracy and precision using quantitative differential interference contrast (DIC) microscopy. Dielectric nanoparticles are detected optically by the conversion of the optical phase change into an intensity change using DIC. Phase images of individual nanoparticles were retrieved from DIC by Wiener filtering, and a quantitative methodology to extract nanoparticle sizes was developed. Using polystyrene beads of 100 nm radius as size standard, we show that the method determines this radius within a few nm accuracy. The smallest detectable polystyrene bead is limited by background and shot-noise, which depend on acquisition and analysis parameters, including the objective numerical aperture, the DIC phase offset, and the refractive index contrast between particles and their surrounding. A sensitivity limit potentially reaching down to 1.8 nm radius was inferred. As application example, individual nanodiamonds with nominal sizes below 50 nm were measured, and were found to have a nearly exponential size distribution with 28 nm mean value. Considering the importance of dielectric nanoparticles in many fields, from naturally occurring virions to polluting nanoplastics, the proposed method could offer a powerful quantitative tool for nanoparticle analysis, combining accuracy, sensitivity and high-throughput with widely available and easy-to-use DIC microscopy

physics.optics↗

arXiv2021

Simultaneous microscopic imaging of thickness and refractive index of thin layers by heterodyne interferometric reflectometry (HiRef)

The detection of spatial or temporal variations in very thin samples has important applications in the biological sciences. For example, cellular membranes exhibit changes in lipid composition and order, which in turn modulate their function in space and time. Simultaneous measurement of thickness and refractive index would be one way to observe these variations, yet doing it noninvasively remains an elusive goal. Here we present a microscopic-imaging technique to simultaneously measure the thickness and refractive index of thin layers in a spatially resolved manner using reflectometry. The heterodyne-detected interference between a light field reflected by the sample and a reference field allows measurement of the amplitude and phase of the reflected field and thus determination of the complex reflection coefficient. Comparing the results with the simulated reflection of a thin layer under coherent illumination of high numerical aperture by the microscope objective, the refractive index and thickness of the layer can be determined. We present results on a layer of polyvinylacetate (PVA) with a thickness of approximately 80~nm. These results have a precision better than 10\% in the thickness and better than 1\% in the refractive index and are consistent within error with measurements by quantitative differential interference contrast (qDIC) and literature values. We discuss the significance of these results, and the possibility of performing accurate measurements on nanometric layers. Notably, the shot-noise limit of the technique is below 0.5~nm in thickness and 0.0005 in refractive index for millisecond measurement times.

physics.optics↗

arXiv2018

Production of Metal-free Diamond Nanoparticles

In this paper, the controlled production of high quality metal-free diamond nanoparticles is demonstrated. Milling with tempered steel is shown to leave behind iron oxide contamination which is difficult to remove. Milling with SiN alleviates this issue but generates more non diamond carbon. Thus the choice of milling materials is critically determined by the acceptable contaminants in the ultimate application. The removal of metal impurities, present in all commercially available nanoparticles, will open new possibilities towards the production of customised diamond nanoparticles, covering the most demanding quantum applications.

cond-mat.mtrl-sci↗

arXiv2018

Long Exciton Dephasing Time and Coherent Phonon Coupling in CsPbBr$_{2}$Cl Perovskite Nanocrystals

Fully-inorganic cesium lead halide perovskite nanocrystals (NCs) have shown to exhibit outstanding optical properties such as wide spectral tunability, high quantum yield, high oscillator strength as well as blinking-free single photon emission and low spectral diffusion. Here, we report measurements of the coherent and incoherent exciton dynamics on the 100 fs to 10 ns timescale, determining dephasing and density decay rates in these NCs. The experiments are performed on CsPbBr$_{2}$Cl NCs using transient resonant three-pulse four-wave mixing (FWM) in heterodyne detection at temperatures ranging from 5 K to 50 K. We found a low-temperature exciton dephasing time of 24.5$\pm$1.0 ps, inferred from the decay of the photon-echo amplitude at 5 K, corresponding to a homogeneous linewidth (FWHM) of 54$\pm$5 μeV. Furthermore, oscillations in the photon-echo signal on a picosecond timescale are observed and attributed to coherent coupling of the exciton to a quantized phonon mode with 3.45 meV energy.

cond-mat.mes-hall↗

arXiv2017

Background-free 3D nanometric localisation and sub-nm asymmetry detection of single plasmonic nanoparticles by four-wave mixing interferometry with optical vortices

Single nanoparticle tracking using optical microscopy is a powerful technique with many applications in biology, chemistry and material sciences. Despite significant advances, localising objects with nanometric position accuracy in a scattering environment remains challenging. Applied methods to achieve contrast are dominantly fluorescence based, with fundamental limits in the emitted photon fluxes arising from the excited-state lifetime as well as photobleaching. Furthermore, every localisation method reported to date requires signal acquisition from multiple spatial points, with consequent speed limitations. Here, we show a new four-wave mixing interferometry technique, whereby the position of a single non-fluorescing gold nanoparticle is determined with better than 20 nm accuracy in plane and 1 nm axially from rapid single-point acquisition measurements by exploiting optical vortices. The technique is also uniquely sensitive to particle asymmetries of only 0.5% aspect ratio, corresponding to a single atomic layer of gold, as well as particle orientation, and the detection is background-free even inside biological cells. This method opens new ways of of unraveling single-particle trafficking within complex 3D architectures.

physics.optics↗

arXiv2014

Evidence of giant oscillator strength in the exciton dephasing of CdSe nanoplatelets measured by resonant four-wave mixing

We measured the intrinsic ground-state exciton dephasing and population dynamics in colloidal quasi two-dimensional (2D) CdSe nanoplatelets at low temperature (5-50K) using transient resonant four-wave mixing in heterodyne detection. Our results indicate that below 20K the exciton dephasing is lifetime limited, with the exciton population lifetime being as fast as 1ps. This is consistent with an exciton lifetime given by a fast radiative decay due to the large in-plane coherence area of the exciton center-of-mass motion in these quasi 2D systems compared to spherical nanocrystals.

cond-mat.mes-hall↗

arXiv2013

Polarization-resolved extinction and scattering cross-section of individual gold nanoparticles measured by wide-field microscopy on a large ensemble

We report a simple, rapid, and quantitative wide-field technique to measure the optical extinction $σ_{\rm ext}$ and scattering $σ_{\rm sca}$ cross-section of single nanoparticles using wide-field microscopy enabling simultaneous acquisition of hundreds of nanoparticles for statistical analysis. As a proof of principle, we measured nominally spherical gold nanoparticles of 40\,nm and 100\,nm diameter and found mean values and standard deviations of $σ_{\rm ext}$ and $σ_{\rm sca}$ consistent with previous literature. Switching from unpolarized to linearly polarized excitation, we measured $σ_{\rm ext}$ as a function of the polarization direction, and used it to characterize the asphericity of the nanoparticles. The method can be implemented cost-effectively on any conventional wide-field microscope and is applicable to any nanoparticles.

cond-mat.mtrl-sci↗

arXiv2012

Femtosecond phase-resolved microscopy of plasmon dynamics in individual gold nanospheres

The selective optical detection of individual metallic nanoparticles (NPs) with high spatial and temporal resolution is a challenging endeavour, yet is key to the understanding of their optical response and their exploitation in applications from miniaturised optoelectronics and sensors to medical diagnostics and therapeutics. However, only few reports on ultrafast pump-probe spectroscopy on single small metallic NPs are available to date. Here, we demonstrate a novel phase-sensitive four-wave mixing (FWM) microscopy in heterodyne detection to resolve for the first time the ultrafast changes of real and imaginary part of the dielectric function of single small (<40nm) spherical gold NPs. The results are quantitatively described via the transient electron temperature and density in gold considering both intraband and interband transitions at the surface plasmon resonance. This novel microscopy technique enables background-free detection of the complex susceptibility change even in highly scattering environments and can be readily applied to any metal nanostructure.

cond-mat.mes-hall↗

arXiv2011

Spin--flip Limited Exciton Dephasing in CdSe/ZnS Colloidal Quantum Dots

The dephasing time of the lowest bright exciton in CdSe/ZnS wurtzite quantum dots is measured from 5 K to 170 K and compared with density dynamics within the exciton fine structure using a sensitive three-beam four-wave-mixing technique unaffected by spectral diffusion. Pure dephasing via acoustic phonons dominates the initial dynamics, followed by an exponential zero-phonon line dephasing of 109 ps at 5 K, much faster than the ~10 ns exciton radiative lifetime. The zero-phonon line dephasing is explained by phonon-assisted spin-flip from the lowest bright state to dark exciton states. This is confirmed by the temperature dependence of the exciton lifetime and by direct measurements of the bright-dark exciton relaxation. Our results give an unambiguous evidence of the physical origin of the exciton dephasing in these nanocrystals.

cond-mat.mes-hall↗