Characterization of different substrates for Raman spectroscopic imaging of eukaryotic cells

For Raman spectroscopic analyses of the cells and other biological samples, the choice of the right substrate material is very important to avoid loss of information in characteristic spectral features because of competing background signals. In the current study, Raman spectroscopy is used to characterize several potential Raman substrates. Raman vibrational bands of the substrate material are discussed. The surface topography is analyzed by atomic force microscopy, and the root mean square surface roughness values are reported. Biocompatibility of the substrates is tested with Hep G2 cells evaluating cellular morphology as well as live/dead staining. Calcium fluoride, silicon, fused silica, borofloat glass, and silicon nitride membranes support cell growth and adherence. Silicon, borofloat glass, and fused silica give rise to Raman signals in the region of interest. Calcium fluoride substrate (UV grade) is suitable for Raman spectroscopic investigation of living cells. Nickel foil is suitable substrate for Raman spectroscopic investigation but cellular adherence and viability depend on the quality of the foil. Silicon nitride membranes coated with nickel chrome is a suitable Raman substrate in closed microfluidic systems. Copyright © 2016 John Wiley & Sons, Ltd.     

Effectiveness of natural and synthetic blocking reagents and their application for detecting food allergens in enzyme-linked immunosorbent assays

Blocking is an important step before an enzyme-linked immunosorbent assay (ELISA) can be performed. It reduces non-specific binding to the microtiter plate to a minimum. For detecting food allergens by means of ELISA, the problem with protein blocking solutions is obvious. The blocker might interfere with the antibodies of the assay and leads to false positive results. Therefore, other blocking solutions are greatly needed. There are some alternatives like synthetic blockers or carbohydrates. Comparisons of these different blocking agents, namely proteins, carbohydrates, and synthetic blockers, were made at different reaction conditions. The incubation periods and temperatures were varied, as well as the pH. The best combinations were evaluated and compared, in respect of their blocking efficiency. The two best non-proteinaceous blockers, i.e. polyvinylalcohol and Ficoll, were subsequently applied to ELISA tests for the determination of α-casein and peanut. The study showed that Ficoll and PVA did as well as BSA in buffer solution. Therefore, they can be considered as alternative blocking reagents for ELISA, especially for the detection of food allergens.     

Compton scattering by the proton through θcms = 75° and 90° in the Δ-resonance region

Differential cross sections for Compton scattering by the proton have been measured in the energy interval between 200 and 500 MeV at scattering angles of θcms = 75° and θcms = 90° using the CATS, the CATS/TRAJAN, and the COPP setups with the Glasgow Tagger at MAMI (Mainz). The data are compared with predictions from dispersion theory using photo-meson amplitudes from the recent VPI solution SM95. The experiment and the theoretical procedure are described in detail. It is found that the experiment and predictions are in agreement as far as the energy dependence of the differential cross sections in the Δ-range is concerned. However, there is evidence that a scaling down of the resonance part of the M1+3/2 photo-meson amplitude by (2.8 ± 0.9)% is required in comparison with the VPI analysis. The deduced value of the M1+3/2-photoproduction amplitude at the resonance energy of 320 MeV is: |M1+3/2| = (39.6 ± 0.4) × 10−3 mπ+−1.     

X-ray triple-crystal diffractometer investigation of arsenic implanted silicon after pulsed laser irradiation

X-ray triple-crystal diffractometry (TCD) of parallel (+n, -n, +n) setting was used to investigate the structural changes of silicon surface layers after arsenic implantation and pulsed laser irradiation. The implated layers of different doses show enlarged lattice constants in the disturbed crystalline parts. After low energy annealing a polycrystalline surface layer is built up from the amorphous layer while the crystalline layer below it is not significantly changed. High energy annealed regions show recrystallisation with high degree of perfection and decreased lattice constant. The results of TCD measurements are compared with results of double-crystal diffractometer investigations and advantages of the TCD method are discussed.     

Integrated waveguide sensor utilizing organic photodiodes

We present a novel optical sensor platform, combining monolithically integrated ring‐like sensor waveguides together with ring‐shaped thin‐film organic photodiodes (OPDs) on one substrate. The OPDs serve as integrated light detectors, simplifying the detection system by minimizing the number of required optical components. The waveguide structures, including a means of coupling light in and out of the waveguides, serve as sensing elements. The functionality of the concept is demonstrated by an integrated carbon dioxide sensor, utilizing absorbance as sensing principle. The integrated optical sensor platform is suitable for the parallel detection of multiple parameters in a single sensor chip using sensor arrays. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Screen-printed polymer waveguides for integrated optics

We demonstrate screen printing of polymer waveguides on rigid and flexible substrates. The waveguides are characterized by optical microscopy and propagation losses are determined by cut-back measurements. Furthermore, the efficiency of light coupling using fluorescent molecules within the waveguides is measured with regard to the application feasibility in sensor systems. It is shown that minimum propagation losses of 0.74 dB/cm can be reached with this low-cost printing technique.     

Photoemission of protons from 4He in the ⊿-resonance region

High resolution proton energy spectra from the ⁴He(γ,p) reaction have been measured with tagged photons in the range E_γ = 130–525 MeV using the large Mainz NaI(Tl) spectrometer at \([Theta _p^{lab} = 37.1^circ ]\) . Three separate reaction channels were identified, viz. ⁴He(γ,p)t two-body breakup, ⁴He photodisintegration via two-nucleon photon absorption processes and the quasifree pion production channel. Differential cross sections are presented for each of these channels as a function of photon energy. The sum of the two-nucleon photon absorption and the quasifree pion production differential cross sections, in the CM system, resembles that of the corresponding free-nucleon differential cross section when Fermi motion is taken into account.     

Hepatic Vitamin A Content Investigation Using Coherent Anti‐Stokes Raman Scattering Microscopy

Standard techniques for examining the distribution of vitamin A in liver either require staining or lead to rapid photobleaching of the molecule. A potentially better alternative approach is to use coherent anti‐Stokes Raman scattering (CARS) microscopy; a fast, label‐free, non‐disruptive imaging method that provides contrast based on molecular vibrations. This contribution evaluates the viability of CARS microscopy for imaging vitamin A within thick hepatic tissue under physiological conditions by tuning into its characteristic vibrational band in the fingerprint region. Additional information about the morphology and architecture of the tissue was acquired using second harmonic generation (SHG) and multi‐photon excited fluorescence (MPEF) to help mapping the intra‐lobular positions of the vitamin A droplets. We demonstrate the capability of our multimodal imaging framework to selectively image lipid‐soluble vitamin A droplets deep in bulk liver tissue with a high contrast while co‐registering a complementary morphological background that clearly visualizes hepatic lobules. The results obtained envisage the good prospect of the technique for in vivo studies assessing vitamin A distribution heterogeneity and how it is affected by the progression of hepatic diseases.     

Commercialized rapid immunoanalytical tests for determination of allergenic food proteins: an overview

Food allergies have become an important health issue especially in industrialized countries. Undeclared allergenic ingredients or the presence of “hidden” allergens because of contamination during the food production process pose great health risks to sensitised individuals. The EU directive for food labelling lists allergenic foods that have to be declared on food products by the manufacturers. The list includes gluten-containing cereals, crustaceans, eggs, fish, peanuts, soybeans, milk, various nuts (e.g. almond, hazelnut, and walnut, etc.), celery, mustard, sesame seeds, lupin, and molluscs. Reliable methods for detection and quantification of food allergens are needed that can be applied in a fast and easy-to-use manner, are portable, and need only limited technical equipment. This review focuses on the latest developments in food allergen analysis with special emphasis on fast immunoanalytical methods such as rapid enzyme-linked immunosorbent assays (ELISA), lateral-flow immunochromatographic assays (LFA) and dipstick tests. Emerging technologies such as immunochemical microarrays and biosensors are also discussed and their application to food allergen analysis is reviewed. Finally, a comprehensive overview of rapid immunochemical test kits that are currently available commercially is given in tabular form.