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Revealing the Chemical Composition of Birch Pollen Grains by Raman Spectroscopic Imaging

2022, Stiebing, Clara, Post, Nele, Schindler, Claudia, Göhrig, Bianca, Lux, Harald, Popp, Jürgen, Heutelbeck, Astrid, Schie, Iwan W.

The investigation of the biochemical composition of pollen grains is of the utmost interest for several environmental aspects, such as their allergenic potential and their changes in growth conditions due to climatic factors. In order to fully understand the composition of pollen grains, not only is an in-depth analysis of their molecular components necessary but also spatial information of, e.g., the thickness of the outer shell, should be recorded. However, there is a lack of studies using molecular imaging methods for a spatially resolved biochemical composition on a single-grain level. In this study, Raman spectroscopy was implemented as an analytical tool to investigate birch pollen by imaging single pollen grains and analyzing their spectral profiles. The imaging modality allowed us to reveal the layered structure of pollen grains based on the biochemical information of the recorded Raman spectra. Seven different birch pollen species collected at two different locations in Germany were investigated and compared. Using chemometric algorithms such as hierarchical cluster analysis and multiple-curve resolution, several components of the grain wall, such as sporopollenin, as well as the inner core presenting high starch concentrations, were identified and quantified. Differences in the concentrations of, e.g., sporopollenin, lipids and proteins in the pollen species at the two different collection sites were found, and are discussed in connection with germination and other growth processes.

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Beyond Beer's Law: Why the Index of Refraction Depends (Almost) Linearly on Concentration

2020, Mayerhöfer, Thomas G., Dabrowska, Alicja, Schwaighofer, Andreas, Lendl, Bernhard, Popp, Jürgen

Beer's empiric law states that absorbance is linearly proportional to the concentration. Based on electromagnetic theory, an approximately linear dependence can only be confirmed for comparably weak oscillators. For stronger oscillators the proportionality constant, the molar attenuation coefficient, is modulated by the inverse index of refraction, which is itself a function of concentration. For comparably weak oscillators, the index of refraction function depends, like absorbance, linearly on concentration. For stronger oscillators, this linearity is lost, except at wavenumbers considerably lower than the oscillator position. In these transparency regions, linearity between the change of the index of refraction and concentration is preserved to a high degree. This can be shown with help of the Kramers–Kronig relations which connect the integrated absorbance to the index of refraction change at lower wavenumbers than the corresponding band. This finding builds the foundation not only for refractive index sensing, but also for new interferometric approaches in IR spectroscopy, which allow measuring the complex index of refraction function. © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.

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Characterization of Zahari Zograph’s nave wall paintings in the church “The nativity of the virgin” of Rila Monastery (Bulgaria) by vibrational spectroscopy and SEM–EDX analysis

2018, Yancheva, Denitsa, Tapanov, Stefan, Velcheva, Evelina, Stamboliyska, Bistra, Glavcheva, Zornitza, Stoyanov, Simeon, Haralampiev, Nikifor, Fischer, Dieter, Lederer, Albena

An analytical study on the nave mural paintings of the church “The Nativity of the Virgin” of Rila monastery, Bulgaria, painted by Zahari Zograph was carried out. Vibrational spectroscopy was applied to identify the pigments and organic materials used in the mural paintings. To complement the spectral information, elemental composition of the samples was determined by SEM-EDX. The data showed that smalt with carbohydrate binder was applied for the blue background, green colour was executed by green earths and red-orange colour – by red lead. Azurite is the pigment used to paint the blue colour of the saints’ hoods. The mordant for gilding was prepared of drying oil, resin and siccative metal oxides as evidenced by SEM-EDX, ATR-FTIR and pyrolysis GC-MS analysis. The use of azurite is related to Zahari Zograph’s works as it was not found in any of the previously studied murals in the church painted by other artists.