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

Ultrafast exciton dephasing in PbS colloidal quantum dots

In this work, we have measured the ground state excitonic dephasing in PbS QDs of sizes from 3.7nm to 5.7nm diameter in the temperature range from 5K to 100K by transient degenerate four-wave mixing (FWM) using 100fs pulses. A combination of heterodyne and k-selection detection was implemented to increase sensitivity and enable 4 orders of magnitude dynamic range in the FWM field detection

Lo Svezzamento nel coniglio: effetti della stagione e dell’età sulla produzione e qualità della carne

Two studies were performed on weaning rabbits: - A six-years investigation was carried out on rabbits reared under constant photoperiod (16L:8D). Pregnancy rate and numerical productivity at weaning appeared to be significantly affected by season. During July and August were observed the lowest number of matings, pregnancy rate and number of weaned subjects. - In order to clarify the influence of weaning age on productive traits of rabbits and of commercial characteristics of carcasses at slaughter, a survey was carried out on 192 crossbred rabbits, 96 weaned at 28 (W28) and 96 at 63 (W63) days of age. Live weight at 83 d and daily weight gain from 64 to 83 d of the 2 groups showed significant differences. W28 rabbits also performed a greater daily solid feed intake and a better feed conversion index. Average dressing out percentage of carcasses was significantly higher in W28 vs W63 as well as loin width, chest width and chest depth measurements. At dissection, meat to bone ratio of W28 carcasses resulted greater than W63. The results also showed the best quality dietary and nutritional properties of meat rabbits W28. In particular, the iron content was higher (P&lt;0.001), and indices of atherogenicity and trombogenicity were more favorable. Weaning age is confirmed to be a considerable variable with reference to the quantitative and qualitative traits of the rabbit meat production.</P

Sparse sampling for fast hyperspectral coherent anti-Stokes Raman scattering imaging

We demonstrate a method to increase the acquisition speed in coherent anti-Stokes Raman scattering (CARS) hyperspectral imaging while retaining the relevant spectral information. The method first determines the important spectral components of a sample from a hyperspectral image over a small number of spatial points but a large number of spectral points covering the accessible spectral range and sampling the instrument spectral resolution at the Nyquist limit. From these components we determine a small set of frequencies needed to retrieve the weights of the components with minimum error for a given measurement noise. Hyperspectral images with a large number of spatial points, for example covering a large spatial region, are then measured at this small set of frequencies, and a reconstruction algorithm is applied to generate the full spectral range and resolution. The resulting spectra are suited to retrieve from the CARS intensity the CARS susceptibility which is linear in the concentration, and apply unsupervised quantitative analysis methods such as FSC3 [1]. We demonstrate the method on CARS hyperspectral images of human osteosarcoma U2OS cell, with a reduction in the acquisition time by a factor of 25. This method is suited also for other coherent vibrational microscopy techniques such as stimulated Raman scattering, and in general for hyperspectral imaging techniques with sequential spectral acquisition

Label-free quantitative chemical imaging and classification analysis of adipogenesis using mouse embryonic stem cells

Stem cells have received much attention recently for their potential utility in regenerative medicine. The identification of their differentiated progeny often requires complex staining procedures, and is challenging for intermediary stages which are a priori unknown. In this work, the ability of label‐free quantitative coherent anti‐Stokes Raman scattering (CARS) micro‐spectroscopy to identify populations of intermediate cell states during the differentiation of murine embryonic stem cells into adipocytes is assessed. Cells were imaged at different days of differentiation by hyperspectral CARS, and images were analysed with an unsupervised factorization algorithm providing Raman‐like spectra and spatially resolved maps of chemical components. Chemical decomposition combined with a statistical analysis of their spatial distributions provided a set of parameters that were used for classification analysis. The first 2 principal components of these parameters indicated 3 main groups, attributed to undifferentiated cells, cells differentiated into committed white pre‐adipocytes, and differentiating cells exhibiting a distinct protein globular structure with adjacent lipid droplets. An unsupervised classification methodology was developed, separating undifferentiated cell from cells in other stages, using a novel method to estimate the optimal number of clusters. The proposed unsupervised classification pipeline of hyperspectral CARS data offers a promising new tool for automated cell sorting in lineage analysis

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

Bessel-beam hyperspectral CARS microscopy with sparse sampling: enabling high-content high-throughput label-free quantitative chemical imaging

Microscopy-based high-content and high-throughput analysis of cellular systems plays a central role in drug discovery. However, for contrast and specificity, the majority of assays require a fluorescent readout which always comes with the risk of alteration of the true biological conditions. In this work, we demonstrate a label-free imaging platform which combines chemically specific hyperspectral coherent anti-Stokes Raman scattering microscopy with sparse sampling and Bessel beam illumination. This enabled us to screen multiwell plates at high speed, while retaining the high-content chemical analysis of hyperspectral imaging. To demonstrate the practical applicability of the method we addressed a critical side effect in drug screens, namely, drug-induced lipid storage within hepatic tissue. We screened 15 combinations of drugs and neutral lipids added to human HepG2 liver cells and developed a high-content quantitative data analysis pipeline which extracted the spectra and spatial distributions of lipid and protein components. We then used their combination to train a support vector machine discriminative algorithm. Classification of the drug responses in terms of phospholipidosis versus steatosis was achieved in a completely label-free assay

Hyperspectral analysis applied to micro-Brillouin maps of amyloid-beta plaques in Alzheimer’s disease brains

A recent investigation on the architecture and chemical composition of amyloid-β (Aβ) plaques in ex vivo histological sections of an Aβ-overexpressing transgenic mouse hippocampus has shed light on the infrared light signature of cell-activation related biomarkers of Alzheimer’s disease. A correlation was highlighted between the biomechanical properties detected by Brillouin microscopy and the molecular make-up of Aβ plaques provided by FTIR spectroscopic imaging and Raman microscopy (with correlative immunofluorescence imaging) in this animal model of the disease. In the Brillouin spectra of heterogeneous materials such as biomedical samples, peaks are likely the result of multiple contributions, more or less overlaid on a spatial and spectral scale. The ability to disentangle these contributions is very important as it may give access to discrete components that would otherwise be buried within the Brillouin peak envelope. Here, we applied an unsupervised non-negative matrix factorization method to analyse the spontaneous Brillouin microscopy maps of Aβ plaques in transgenic mouse hippocampal sections. The method has already been proven successful in decomposing chemical images and is applied here for the first time to acoustic maps acquired with a Fabry–Perot Brillouin microscope. We extracted and visualised a decrease in tissue rigidity from the core through to the periphery of the plaque, with spatially distinct components that we assigned to specific entities. This work demonstrates that it is possible to reveal the structure and mechanical properties of Aβ plaques, with details visualized by the projection of the mechanical contrast into a few relevant channels

Photochemical approach for multiplexed biofunctionalisation of gallium arsenide

The optoelectronic properties of gallium arsenide (GaAs) hold great promise in biosensing applications, currently being held back by the lack of methodologies reporting the spatially selective functionalisation of this material with multiple biomolecules. Here, we exploit the use of a photoreactive crosslinker - a diazirine derivative - for spatially selective covalent immobilisation of multiple bioreceptors on the GaAs surface. As a proof of principle we show the immobilisation of two proteins: neutravidin and endosulfine alpha protein. X-ray photoelectron spectroscopy results showed the presence of the biomolecules on the GaAs regions selectively exposed to ultraviolet light. The approach presented here is applicable to the covalent attachment of other biomolecules, paving the way for using GaAs as a platform for multiplexed biosensing

Wavelength agile quantum dot laser for lab-on-chip optical biosensors

We report simulations of photonic, optoelectronic, and thermal properties of a wavelength agile photonic crystal laser for lab-on-chip optical biosensors. We find that with an optimized cavity, lasing can be achieved with a single layer of QDs, providing a tunability by self-heating over 5 nm