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- ItemBiochemical Characterization of Mouse Retina of an Alzheimer's Disease Model by Raman Spectroscopy(Washington, DC : ACS Publications, 2020) Stiebing, Clara; Jahn, Izabella J.; Schmitt, Michael; Keijzer, Nanda; Kleemann, Robert; Kiliaan, Amanda J.; Drexler, Wolfgang; Leitgeb, Rainer A.; Popp, JürgenThe presence of biomarkers characteristic for Alzheimer's disease in the retina is a controversial topic. Raman spectroscopy offers information on the biochemical composition of tissues. Thus, it could give valuable insight into the diagnostic value of retinal analysis. Within the present study, retinas of a double transgenic mouse model, that expresses a chimeric mouse/human amyloid precursor protein and a mutant form of human presenilin 1, and corresponding control group were subjected to ex vivo Raman imaging. The Raman data recorded on cross sections of whole eyes highlight the layered structure of the retina in a label-free manner. Based on the Raman information obtained from en face mounted retina samples, a discrimination between healthy and Alzheimer's disease retinal tissue can be done with an accuracy of 85.9%. For this a partial least squares-linear discriminant analysis was applied. Therefore, although no macromolecular changes in form of, i.e., amyloid beta plaques, can be noticed based on Raman spectroscopy, subtle biochemical changes happening in the retina could lead to Alzheimer's disease identification. ©
- ItemShape-Memory Metallopolymers Based on Two Orthogonal Metal–Ligand Interactions(Weinheim : Wiley-VCH, 2021) Meurer, Josefine; Hniopek, Julian; Bätz, Thomas; Zechel, Stefan; Enke, Marcel; Vitz, Jürgen; Schmitt, Michael; Popp, Jürgen; Hager, Martin D.; Schubert, Ulrich S.A new shape-memory polymer is presented, in which both the stable phase as well as the switching unit consist of two different metal complexes. Suitable metal ions, which simultaneously form labile complexes with histidine and stable ones with terpyridine ligands, are identified via isothermal titration calorimetry (ITC) measurements. Different copolymers are synthesized, which contain butyl methacrylate as the main monomer and the metal-binding ligands in the side chains. Zn(TFMS)2 and NiCl2 are utilized for the dual crosslinking, resulting in the formation of metallopolymer networks. The switching temperature can simply be tuned by changing the composition as well as by the choice of the metal ion. Strain fixity rates (about 99%) and very high strain recovery rates (up to 95%) are achieved and the mechanism is revealed using different techniques such as Raman spectroscopy. © 2021 The Authors. Advanced Materials published by Wiley-VCH GmbH
- ItemStealth Effect of Short Polyoxazolines in Graft Copolymers: Minor Changes of Backbone End Group Determine Liver Cell-Type Specificity(Washington, DC : ACS Publications, 2021) Muljajew, Irina; Huschke, Sophie; Ramoji, Anuradha; Cseresnyés, Zoltán; Hoeppener, Stephanie; Nischang, Ivo; Foo, Wanling; Popp, Jürgen; Figge, Marc Thilo; Weber, Christine; Bauer, Michael; Schubert, Ulrich S.; Press, Adrian T.Dye-loaded micelles of 10 nm diameter formed from amphiphilic graft copolymers composed of a hydrophobic poly(methyl methacrylate) backbone and hydrophilic poly(2-ethyl-2-oxazoline) side chains with a degree of polymerization of 15 were investigated concerning their cellular interaction and uptake in vitro as well as their interaction with local and circulating cells of the reticuloendothelial system in the liver by intravital microscopy. Despite the high molar mass of the individual macromolecules (Mn ≈ 20 kg mol-1), backbone end group modification by attachment of a hydrophilic anionic fluorescent probe strongly affected the in vivo performance. To understand these effects, the end group was additionally modified by the attachment of four methacrylic acid repeating units. Although various micelles appeared similar in dynamic light scattering and cryo-transmission electron microscopy, changes in the micelles were evident from principal component analysis of the Raman spectra. Whereas an efficient stealth effect was found for micelles formed from polymers with anionically charged or thiol end groups, a hydrophobic end group altered the micelles' structure sufficiently to adapt cell-type specificity and stealth properties in the liver. © 2021 The Authors. Published by American Chemical Society.
- ItemBeyond Beer's Law: Revisiting the Lorentz-Lorenz Equation(Weinheim : Wiley-VCH Verl., 2020) Mayerhöfer, Thomas G.; Popp, JürgenIn this contribution we show how the Lorentz-Lorenz and the Clausius-Mosotti equations are related to Beer's law. Accordingly, the linear concentration dependence of absorbance is a consequence of neglecting the difference between the local and the applied electric field. Additionally, it is necessary to assume that the absorption index and the related refractive index change is small. By connecting the Lorentz-Lorenz equations with dispersion theory, it becomes obvious that the oscillators are coupled via the local field. We investigate this coupling with numerical examples and show that, as a consequence, the integrated absorbance of a single band is in general no longer linearly depending on the concentration. In practice, the deviations from Beer's law usually do not set in before the density reaches about one tenth of that of condensed matter. For solutions, the Lorentz-Lorenz equations predict a strong coupling also between the oscillators of solute and solvent. In particular, in the infrared spectral region, the absorption coefficients are prognosticated to be much higher due to this coupling compared to those in the gas phase. © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.
- ItemEosinophils and Neutrophils-Molecular Differences Revealed by Spontaneous Raman, CARS and Fluorescence Microscopy(Basel : MDPI, 2020) Dorosz, Aleksandra; Grosicki, Marek; Dybas, Jakub; Matuszyk, Ewelina; Rodewald, Marko; Meyer, Tobias; Popp, Jürgen; Malek, Kamilla; Baranska, MalgorzataLeukocytes are a part of the immune system that plays an important role in the host's defense against viral, bacterial, and fungal infections. Among the human leukocytes, two granulocytes, neutrophils (Ne) and eosinophils (EOS) play an important role in the innate immune system. For that purpose, eosinophils and neutrophils contain specific granules containing protoporphyrin-type proteins such as eosinophil peroxidase (EPO) and myeloperoxidase (MPO), respectively, which contribute directly to their anti-infection activity. Since both proteins are structurally and functionally different, they could potentially be a marker of both cells' types. To prove this hypothesis, UV-Vis absorption spectroscopy and Raman imaging were applied to analyze EPO and MPO and their content in leukocytes isolated from the whole blood. Moreover, leukocytes can contain lipidic structures, called lipid bodies (LBs), which are linked to the regulation of immune responses and are considered to be a marker of cell inflammation. In this work, we showed how to determine the number of LBs in two types of granulocytes, EOS and Ne, using fluorescence and coherent anti-Stokes Raman scattering (CARS) microscopy. Spectroscopic differences of EPO and MPO can be used to identify these cells in blood samples, while the detection of LBs can indicate the cell inflammation process.
- ItemNovel Biobased Self-Healing Ionomers Derived from Itaconic Acid Derivates(Weinheim : Wiley-VCH, 2021) Meurer, Josefine; Hniopek, Julian; Dahlke, Jan; Schmitt, Michael; Popp, Jürgen; Zechel, Stefan; Hager, Martin D.This article presents novel biobased ionomers featuring self-healing abilities. These smart materials are synthesized from itaconic acid derivates. Large quantities of itaconic acid can be produced from diverse biomass like corn, rice, and others. This study presents a comprehensive investigation of their thermal and mechanical properties via differential scanning calorimetry (DSC), thermo gravimetric analysis (TGA), and FT-Raman and FT-IR measurements as well as dynamic mechanic analysis. Within all these measurements, different kinds of structure-property relationships could be derived from these measurements. For example, the proportion of ionic groups enormously influences the self-healing efficiency. The investigation of the self-healing abilities reveals healing efficiencies up to 99% in 2 h at 90 °C for the itaconic acid based ionomer with the lowest ionic content. © 2020 The Authors. Macromolecular Rapid Communications published by Wiley-VCH GmbH
- ItemFLIm and raman spectroscopy for investigating biochemical changes of bovine pericardium upon genipin cross-linking(Basel : MDPI, 2020) Shaik, Tanveer Ahmed; Alfonso-Garcia, Alba; Richter, Martin; Korinth, Florian; Krafft, Christoph; Marcu, Laura; Popp, JürgenBiomaterials used in tissue engineering and regenerative medicine applications benefit from longitudinal monitoring in a non-destructive manner. Label-free imaging based on fluorescence lifetime imaging (FLIm) and Raman spectroscopy were used to monitor the degree of genipin (GE) cross-linking of antigen-removed bovine pericardium (ARBP) at three incubation time points (0.5, 1.0, and 2.5 h). Fluorescence lifetime decreased and the emission spectrum redshifted compared to that of uncross-linked ARBP. The Raman signature of GE-ARBP was resonance-enhanced due to the GE cross-linker that generated new Raman bands at 1165, 1326, 1350, 1380, 1402, 1470, 1506, 1535, 1574, 1630, 1728, and 1741 cm-1. These were validated through density functional theory calculations as cross-linker-specific bands. A multivariate multiple regression model was developed to enhance the biochemical specificity of FLIm parameters fluorescence intensity ratio (R2 = 0.92) and lifetime (R2 = 0.94)) with Raman spectral results. FLIm and Raman spectroscopy detected biochemical changes occurring in the collagenous tissue during the cross-linking process that were characterized by the formation of a blue pigment which affected the tissue fluorescence and scattering properties. In conclusion, FLIm parameters and Raman spectroscopy were used to monitor the degree of cross-linking non-destructively. © 2020 by the authors. Licensee MDPI, Basel, Switzerland.
- ItemA Machine Learning-Based Raman Spectroscopic Assay for the Identification of Burkholderia mallei and Related Species(Basel : MDPI, 2019) Silge, Anja; Moawad, Amira A.; Bocklitz, Thomas; Fischer, Katja; Rösch, Petra; Roesler, Uwe; Elschner, Mandy C.; Popp, Jürgen; Neubauer, HeinrichBurkholderia (B.) mallei, the causative agent of glanders, and B. pseudomallei, the causative agent of melioidosis in humans and animals, are genetically closely related. The high infectious potential of both organisms, their serological cross-reactivity, and similar clinical symptoms in human and animals make the differentiation from each other and other Burkholderia species challenging. The increased resistance against many antibiotics implies the need for fast and robust identification methods. The use of Raman microspectroscopy in microbial diagnostic has the potential for rapid and reliable identification. Single bacterial cells are directly probed and a broad range of phenotypic information is recorded, which is subsequently analyzed by machine learning methods. Burkholderia were handled under biosafety level 1 (BSL 1) conditions after heat inactivation. The clusters of the spectral phenotypes and the diagnostic relevance of the Burkholderia spp. were considered for an advanced hierarchical machine learning approach. The strain panel for training involved 12 B. mallei, 13 B. pseudomallei and 11 other Burkholderia spp. type strains. The combination of top- and sub-level classifier identified the mallei-complex with high sensitivities (>95%). The reliable identification of unknown B. mallei and B. pseudomallei strains highlighted the robustness of the machine learning-based Raman spectroscopic assay. © 2019 by the authors
- ItemLiquid-Core Microstructured Polymer Optical Fiber as Fiber-Enhanced Raman Spectroscopy Probe for Glucose Sensing(Washington, DC : OSA, 2020) Azkune, Mikel; Frosch, Timea; Arrospide, Eneko; Aldabaldetreku, Gotzon; Bikandi, Iñaki; Zubia, Joseba; Popp, Jürgen; Frosch, TorstenThis work reports the development and application of two liquid-core microstructured polymer optical fibers (LC-mPOF) with different microstructure sizes. They are used in a fiber-enhanced Raman spectroscopy sensing platform, with the aim of detecting glucose in aqueous solutions in the clinically relevant range for sodium-glucose cotransporter 2 inhibitor therapy. The sensing platform is tested for low-concentration glucose solutions using each LC-mPOF. Results confirm that a significant enhancement of the Raman signal is achieved in comparison to conventional Raman spectroscopy. Additional measurements are carried out to obtain the valid measurement range, the resolution, and the limit of detection, showing that the LC-mPOF with 66-µm-diameter central hollow core has the highest potential for future clinical applications. Finally, preliminary tests successfully demonstrate glucose identification in urine. © 1983-2012 IEEE.
- ItemSurface enhanced Raman spectroscopy-based evaluation of the membrane protein composition of the organohalide-respiring Sulfurospirillum multivorans(Chichester [u.a.] : Wiley, 2021) Cialla-May, Dana; Gadkari, Jennifer; Winterfeld, Andreea; Hübner, Uwe; Weber, Karina; Diekert, Gabriele; Schubert, Torsten; Goris, Tobias; Popp, JürgenBacteria often employ different respiratory chains that comprise membrane proteins equipped with various cofactors. Monitoring the protein inventory that is present in the cells under a given cultivation condition is often difficult and time-consuming. One example of a metabolically versatile bacterium is the microaerophilic organohalide-respiring Sulfurospirillum multivorans. Here, we used surface enhanced Raman spectroscopy (SERS) to quickly identify the cofactors involved in the respiration of S. multivorans. We cultured the organism with either tetrachloroethene (perchloroethylene, PCE), fumarate, nitrate, or oxygen as electron acceptors. Because the corresponding terminal reductases of the four different respiratory chains harbor different cofactors, specific fingerprint signals in SERS were expected. Silver nanostructures fabricated by means of electron beam lithography were coated with the membrane fractions extracted from the four S. multivorans cultivations, and SERS spectra were recorded. In the case of S. multivorans cultivated with PCE, the recorded SERS spectra were dominated by Raman peaks specific for Vitamin B12. This is attributed to the high abundance of the PCE reductive dehalogenase (PceA), the key enzyme in PCE respiration. After cultivation with oxygen, fumarate, or nitrate, no Raman spectral features of B12 were found. © 2020 The Authors. Journal of Raman Spectroscopy published by John Wiley & Sons Ltd