Protein clustering in chemically stressed HeLa cells studied by infrared nanospectroscopy

Photo-Thermal Induced Resonance (PTIR) nanospectroscopy, tuned towards amide-I absorption, was used to study the distribution of proteic material in 34 different HeLa cells, of which 18 were chemically stressed by oxidative stress with Na3AsO3. The cell nucleus was found to provide a weaker amide-I signal than the surrounding cytoplasm, while the strongest PTIR signal comes from the perinuclear region. AFM topography shows that the cells exposed to oxidative stress undergo a volume reduction with respect to the control cells, through an accumulation of the proteic material around and above the nucleus. This is confirmed by the PTIR maps of the cytoplasm, where the pixels providing a high amide-I signal were identified with a space resolution of ∼300 × 300 nm. By analyzing their distribution with two different statistical procedures we found that the probability to find protein clusters smaller than 0.6 μm in the cytoplasm of stressed HeLa cells is higher by 35% than in the control cells. These results indicate that it is possible to study proteic clustering within single cells by label-free optical nanospectroscopy

Tunability and Losses of Mid-infrared Plasmonics in Heavily Doped Germanium Thin Films

Heavily-doped semiconductor films are very promising for application in mid-infrared plasmonic devices because the real part of their dielectric function is negative and broadly tunable in this wavelength range. In this work we investigate heavily n-type doped germanium epilayers grown on different substrates, in-situ doped in the $10^{17}$ to $10^{19}$ cm$^{-3}$ range, by infrared spectroscopy, first principle calculations, pump-probe spectroscopy and dc transport measurements to determine the relation between plasma edge and carrier density and to quantify mid-infrared plasmon losses. We demonstrate that the unscreened plasma frequency can be tuned in the 400 - 4800 cm$^{-1}$ range and that the average electron scattering rate, dominated by scattering with optical phonons and charged impurities, increases almost linearly with frequency. We also found weak dependence of losses and tunability on the crystal defect density, on the inactivated dopant density and on the temperature down to 10 K. In films where the plasma was optically activated by pumping in the near-infrared, we found weak but significant dependence of relaxation times on the static doping level of the film. Our results suggest that plasmon decay times in the several-picosecond range can be obtained in n-type germanium thin films grown on silicon substrates hence allowing for underdamped mid-infrared plasma oscillations at room temperature.Comment: 18 pages, 10 figure

Confocal Imaging at 0.3 THz with depth resolution of a painted wood artwork for the identification of buried thin metal foils

A compact confocal terahertz microscope working at 0.30 THz based on all-solid-state components is used to locate buried thin metal foils in a painted wood artwork. Metal foils are used for decoration, and their precise localization under the pictorial layer is relevant information for conservation scientists and restorers, which can neither be obtained by X-ray radiography nor by spectroscopic imaging in the infrared, as we directly show here. The confocal microscopy principle based on the spatial pinhole concept is here implemented by positioning the first focus of an ellipsoidal reflector at the phase center of horn antennas coupled to Schottky diode detector and emitter mounted in rectangular waveguide blocks, together with an optical beamsplitter. The second focus of the reflector is mechanically scanned inside the sample in three dimensions. The predictions of diffraction theory for a confocal microscope at an imaging wavelength of 1.00 mm with numerical aperture of 0.53 are verified experimentally (1.2 and 2.8 mm for the lateral and the axial resolution, respectively). These values of resolution allow a precise determination of the position of buried metal foils in an ancient piece of art hence making restoration interventions possible

Low-temperature stability and sensing performance of mid-infrared bloch surface waves on a one-dimensional photonic crystal

The growing need for new and reliable surface sensing methods is arousing interest in the electromagnetic excitations of ultrathin films, i.e., to generate electromagnetic field distributions that resonantly interact with the most significant quasi-particles of condensed matter. In such a context, Bloch surface waves turned out to be a valid alternative to surface plasmon polaritons to implement high-sensitivity sensors in the visible spectral range. Only in the last few years, however, has their use been extended to infrared wavelengths, which represent a powerful tool for detecting and recognizing molecular species and crystalline structures. In this work, we demonstrate, by means of high-resolution reflectivity measurements, that a one-dimensional photonic crystal can sustain Bloch surface waves in the infrared spectral range from room temperature down to 10 K. To the best of our knowledge, this is the first demonstration of infrared Bloch surface waves at cryogenic temperatures. Furthermore, by exploiting the enhancement of the surface state and the high brilliance of infrared synchrotron radiation, we demonstrate that the proposed BSW-based sensor has a sensitivity on the order of 2.9 cm–1 for each nanometer-thick ice layer grown on its surface below 150 K. In conclusion, we believe that Bloch surface wave-based sensors are a valid new class of surface mode-based sensors for applications in materials science

IgE to cross-reactive carbohydrate determinants (CCD) in childhood: Prevalence, risk factors, putative origins

background IgE antibodies to cross-reactive carbohydrate determinants (CCD) are usually clinically irrelevant but they can be a cause of false positive outcomes of allergen-specific IgE tests in vitro. their prevalence and levels have been so far cross-sectionally examined among adult allergic patients and much less is known about their origins and relevance in childhood. methods we examined CCD with a cross-sectional approach in 1263 Italian pollen allergic children (panallergen in paediatrics, PAN-PED), as well as with a longitudinal approach in 612 german children (Multicenter allergy Study, MAS), whose cutaneous and IgE sensitization profile to a broad panel of allergen extracts and molecules was already known. the presence and levels of IgE to CCD were examined in the sera of both cohorts using bromelain (MUXF3) as reagent and a novel chemiluminescence detection system, operating in a solid phase of fluorescently labelled and streptavidin-coated paramagnetic microparticles (NOVEOS, HYCOR, USA). resultsIgE to CCD was found in 22% of the Italian pollen allergic children, mainly in association with an IgE response to grass pollen. children with IgE to CCD had higher total IgE levels and were sensitized to more allergenic molecules of Phleum pratense than those with no IgE to CCD. among participants of the German MAS birth cohort study, IgE to CCD emerged early in life (even at pre-school age), with IgE sensitization to group 1 and 4 allergen molecules of grasses, and almost invariably persisted over the full observation period.ConclusionsOur results contribute to dissect the immunological origins, onset, evolution and risk factors of CCD-sIgE response in childhood, and raise the hypothesis that group 1 and/or 4 allergen molecules of grass pollen are major inducers of these antibodies through an antigen-specific, T-B cell cognate interaction.IgE antibodies to CCD in childhood, tested in the PAN-PED (cross-sectional approach) and MAS (longitudinal approach) cohorts, are as follows: (1) very frequent in patients with pollen allergy (22%, 275/1263), (2) associated with strong and broad IgE response to grass pollen, (3) emerging with IgE sensitization to grass group 1 and 4 allergens, (4) can start very early in life (positive IgE seen in second year of life) and (5) once started, are almost invariably persistent.imag

Performance Assessment in Fingerprinting and Multi Component Quantitative NMR Analyses

An interlaboratory comparison (ILC) was organized with the aim to set up quality control indicators suitable for multicomponent quantitative analysis by nuclear magnetic resonance (NMR) spectroscopy. A total of 36 NMR data sets (corresponding to 1260 NMR spectra) were produced by 30 participants using 34 NMR spectrometers. The calibration line method was chosen for the quantification of a five-component model mixture. Results show that quantitative NMR is a robust quantification tool and that 26 out of 36 data sets resulted in statistically equivalent calibration lines for all considered NMR signals. The performance of each laboratory was assessed by means of a new performance index (named Qp-score) which is related to the difference between the experimental and the consensus values of the slope of the calibration lines. Laboratories endowed with a Qp-score falling within the suitable acceptability range are qualified to produce NMR spectra that can be considered statistically equivalent in terms of relative intensities of the signals. In addition, the specific response of nuclei to the experimental excitation/relaxation conditions was addressed by means of the parameter named NR. NR is related to the difference between the theoretical and the consensus slopes of the calibration lines and is specific for each signal produced by a well-defined set of acquisition parameters