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- ItemcellSTORM - Cost-effective Super-Resolution on a Cellphone using dSTORM(San Francisco : Public Library of Science, 2019) Diederich, Benedict; Then, Patrick; Jügler, Alexander; Förster, Ronny; Heintzmann, RainerHigh optical resolution in microscopy usually goes along with costly hardware components, such as lenses, mechanical setups and cameras. Several studies proved that Single Molecular Localization Microscopy can be made affordable, relying on off-the-shelf optical components and industry grade CMOS cameras. Recent technological advantages have yielded consumer-grade camera devices with surprisingly good performance. The camera sensors of smartphones have benefited of this development. Combined with computing power smartphones provide a fantastic opportunity for “imaging on a budget”. Here we show that a consumer cellphone is capable of optical super-resolution imaging by (direct) Stochastic Optical Reconstruction Microscopy (dSTORM), achieving optical resolution better than 80 nm. In addition to the use of standard reconstruction algorithms, we used a trained image-to-image generative adversarial network (GAN) to reconstruct video sequences under conditions where traditional algorithms provide sub-optimal localization performance directly on the smartphone. We believe that “cellSTORM” paves the way to make super-resolution microscopy not only affordable but available due to the ubiquity of cellphone cameras.High optical resolution in microscopy usually goes along with costly hardware components, such as lenses, mechanical setups and cameras. Several studies proved that Single Molecular Localization Microscopy can be made affordable, relying on off-the-shelf optical components and industry grade CMOS cameras. Recent technological advantages have yielded consumer-grade camera devices with surprisingly good performance. The camera sensors of smartphones have benefited of this development. Combined with computing power smartphones provide a fantastic opportunity for “imaging on a budget”. Here we show that a consumer cellphone is capable of optical super-resolution imaging by (direct) Stochastic Optical Reconstruction Microscopy (dSTORM), achieving optical resolution better than 80 nm. In addition to the use of standard reconstruction algorithms, we used a trained image-to-image generative adversarial network (GAN) to reconstruct video sequences under conditions where traditional algorithms provide sub-optimal localization performance directly on the smartphone. We believe that “cellSTORM” paves the way to make super-resolution microscopy not only affordable but available due to the ubiquity of cellphone cameras.
- ItemTm3+/Ho3+ co-doped germanate glass double-clad fiber for broadband emission and lasing above 2 µm(Washington D.C. : Optical Society of America, 2019) Kochanowicz, Marcin; Zmojda, Jacek; Miluski, Piotr; Baranowska, Agata; Leich, Martin; Schwuchow, Anka; Jäger, Matthias; Kuwik, M.; Pisarska, Johanna; Pisarski, Wojciech A.; Dorosz, DominikIn this paper, a 2 µm broadband emission under 796 nm laser diode excitation in low phonon energy GeO2-Ga2O3-BaO glass system is co-doped with 0.7Tm2O3/(0.07-0.7)Ho2O3 (mol%). The widest emission band (where the Tm3+ → Ho3+ energy transfer efficiency is 63%) was obtained for 0.7Tm2O3/0.15Ho2O3 co-doped glass from which a double-clad optical fiber was realized and investigated. Optimization of Tm3+/Ho3+ concentration enabled the acquisition of broadband amplified spontaneous emission (ASE) in double-clad optical fiber with a full width at half maximum (FWHM): 377 nm and 662 nm for 3 dB and 10 dB bandwidth, respectively. ASE spectrum is a result of the superposition of (Tm3+: 3H4 →Η3F4) 1.45 µm, (Tm3+: 3F4 → 3H6) 1.8 µm and (Ho3+:5I7 → 5I8) 2 µm emission bands. Hence, highly rare-earth co-doped germanate glass is characterized by a remarkably broader ASE spectrum than silica and tellurite fibers showed promising lasing properties for their further application in tunable and dual wavelength lasers.In this paper, a 2 µm broadband emission under 796 nm laser diode excitation in low phonon energy GeO2-Ga2O3-BaO glass system is co-doped with 0.7Tm2O3/(0.07-0.7)Ho2O3 (mol%). The widest emission band (where the Tm3+ → Ho3+ energy transfer efficiency is 63%) was obtained for 0.7Tm2O3/0.15Ho2O3 co-doped glass from which a double-clad optical fiber was realized and investigated. Optimization of Tm3+/Ho3+ concentration enabled the acquisition of broadband amplified spontaneous emission (ASE) in double-clad optical fiber with a full width at half maximum (FWHM): 377 nm and 662 nm for 3 dB and 10 dB bandwidth, respectively. ASE spectrum is a result of the superposition of (Tm3+: 3H4 →Η3F4) 1.45 µm, (Tm3+: 3F4 → 3H6) 1.8 µm and (Ho3+:5I7 → 5I8) 2 µm emission bands. Hence, highly rare-earth co-doped germanate glass is characterized by a remarkably broader ASE spectrum than silica and tellurite fibers showed promising lasing properties for their further application in tunable and dual wavelength lasers.
- ItemCounterfeit and substandard test of the antimalarial tablet Riamet® by means of Raman hyperspectral multicomponent analysis(Basel : MDPI, 2019) Frosch, Timea; Wyrwich, Elisabeth; Yan, Di; Domes, Christian; Domes, Robert; Popp, Jürgen; Frosch, TorstenThe fight against counterfeit pharmaceuticals is a global issue of utmost importance, as failed medication results in millions of deaths every year. Particularly affected are antimalarial tablets. A very important issue is the identification of substandard tablets that do not contain the nominal amounts of the active pharmaceutical ingredient (API), and the differentiation between genuine products and products without any active ingredient or with a false active ingredient. This work presents a novel approach based on fiber-array based Raman hyperspectral imaging to qualify and quantify the antimalarial APIs lumefantrine and artemether directly and non-invasively in a tablet in a time-efficient way. The investigations were carried out with the antimalarial tablet Riamet® and self-made model tablets, which were used as examples of counterfeits and substandard. Partial least-squares regression modeling and density functional theory calculations were carried out for quantification of lumefantrine and artemether and for spectral band assignment. The most prominent differentiating vibrational signatures of the APIs were presented.
- ItemMicrofluidic Cultivation and Laser Tweezers Raman Spectroscopy of E. coli under Antibiotic Stress(Basel : MDPI, 2018) Pilát, Zdeněk; Bernatová, Silvie; Ježek, Jan; Kirchhoff, Johanna; Tannert, Astrid; Neugebauer, Ute; Samek, Ota; Zemánek, PavelAnalyzing the cells in various body fluids can greatly deepen the understanding of the mechanisms governing the cellular physiology. Due to the variability of physiological and metabolic states, it is important to be able to perform such studies on individual cells. Therefore, we developed an optofluidic system in which we precisely manipulated and monitored individual cells of Escherichia coli. We tested optical micromanipulation in a microfluidic chamber chip by transferring individual bacteria into the chambers. We then subjected the cells in the chambers to antibiotic cefotaxime and we observed the changes by using time-lapse microscopy. Separately, we used laser tweezers Raman spectroscopy (LTRS) in a different micro-chamber chip to manipulate and analyze individual cefotaxime-treated E. coli cells. Additionally, we performed conventional Raman micro-spectroscopic measurements of E. coli cells in a micro-chamber. We found observable changes in the cellular morphology (cell elongation) and in Raman spectra, which were consistent with other recently published observations. The principal component analysis (PCA) of Raman data distinguished between the cefotaxime treated cells and control. We tested the capabilities of the optofluidic system and found it to be a reliable and versatile solution for this class of microbiological experiments.
- ItemAmyloids: From molecular structure to mechanical properties(Amsterdam [u.a.] : Elsevier, 2013) Schleeger, M.; Vandenakker, C.C.; Deckert-Gaudig, T.; Deckert, V.; Velikov, K.P.; Koenderink, G.; Bonn, M.Many proteins of diverse sequence, structure and function self-assemble into morphologically similar fibrillar aggregates known as amyloids. Amyloids are remarkable polymers in several respects. First of all, amyloids can be formed from proteins with very different amino acid sequences; the common denominator is that the individual proteins constituting the amyloid fold predominantly into a β-sheet structure. Secondly, the formation of the fibril occurs through non-covalent interactions between primarily the β-sheets, causing the monomers to stack into fibrils. The fibrils are remarkably robust, considering that the monomers are bound non-covalently. Finally, a common characteristic of fibrils is their unbranched, straight, fiber-like structure arising from the intertwining of the multiple β-sheet filaments. These remarkably ordered and stable nanofibrils can be useful as building blocks for protein-based functional materials, but they are also implicated in severe neurodegenerative diseases. The overall aim of this article is to highlight recent efforts aimed at obtaining insights into amyloid proteins on different length scales. Starting from molecular information on amyloids, single fibril properties and mechanical properties of networks of fibrils are described. Specifically, we focus on the self-assembly of amyloid protein fibrils composed of peptides and denatured model proteins, as well as the influence of inhibitors of fibril formation. Additionally, we will demonstrate how the application of recently developed vibrational spectroscopic techniques has emerged as a powerful approach to gain spatially resolved information on the structure-function relation of amyloids. While spectroscopy provides information on local molecular conformations and protein secondary structure, information on the single fibril level has been developed by diverse microscopic techniques. The approaches to reveal basic mechanical properties of single fibrils like bending rigidity, shear modulus, ultimate tensile strength and fracture behavior are illustrated. Lastly, mechanics of networks of amyloid fibrils, typically forming viscoelastic gels are outlined, with a focus on (micro-) rheological properties. The resulting fundamental insights are essential for the rational design of novel edible and biodegradable protein-based polymers, but also to devise therapeutic strategies to combat amyloid assembly and accumulation during pathogenic disorders.
- ItemCarbon chloride-core fibers for soliton mediated supercontinuum generation(Washington D.C. : Optical Society of America, 2018) Chemnitz, Mario; Gaida, Christian; Gebhardt, Martin; Stutzki, Fabian; Kobelke, Jens; Tünnermann, Andreas; Limpert, Jens; Schmidt, Markus A.We report on soliton-fission mediated infrared supercontinuum generation in liquid-core step-index fibers using highly transparent carbon chlorides (CCl4, C2Cl4). By developing models for the refractive index dispersions and nonlinear response functions, dispersion engineering and pumping with an ultrafast thulium fiber laser (300 fs) at 1.92 μm, distinct soliton fission and dispersive wave generation was observed, particularly in the case of tetrachloroethylene (C2Cl4). The measured results match simulations of both the generalized and a hybrid nonlinear Schrödinger equation, with the latter resembling the characteristics of non-instantaneous medium via a static potential term and representing a simulation tool with substantially reduced complexity. We show that C2Cl4 has the potential for observing non-instantaneous soliton dynamics along meters of liquid-core fiber opening a feasible route for directly observing hybrid soliton dynamics.
- ItemNanostructured Silicon Matrix for Materials Engineering(Weinheim : Wiley-VCH, 2023) Liu, Poting; Schleusener, Alexander; Zieger, Gabriel; Bochmann, Arne; van Spronsen, Matthijs A.; Sivakov, VladimirTin-containing layers with different degrees of oxidation are uniformly distributed along the length of silicon nanowires formed by a top-down method by applying metalorganic chemical vapor deposition. The electronic and atomic structure of the obtained layers is investigated by applying nondestructive surface-sensitive X-ray absorption near edge spectroscopy using synchrotron radiation. The results demonstrated, for the first time, a distribution effect of the tin-containing phases in the nanostructured silicon matrix compared to the results obtained for planar structures at the same deposition temperatures. The amount and distribution of tin-containing phases can be effectively varied and controlled by adjusting the geometric parameters (pore diameter and length) of the initial matrix of nanostructured silicon. Due to the occurrence of intense interactions between precursor molecules and decomposition by-products in the nanocapillary, as a consequence of random thermal motion of molecules in the nanocapillary, which leads to additional kinetic energy and formation of reducing agents, resulting in effective reduction of tin-based compounds to a metallic tin state for molecules with the highest penetration depth in the nanostructured silicon matrix. This effect will enable clear control of the phase distributions of functional materials in 3D matrices for a wide range of applications.
- ItemResonance Raman Spectro-Electrochemistry to Illuminate Photo-Induced Molecular Reaction Pathways(Basel : MDPI, 2019) Zedler, Linda; Krieck, Sven; Kupfer, Stephan; Dietzek, BenjaminElectron transfer reactions play a key role for artificial solar energy conversion, however, the underlying reaction mechanisms and the interplay with the molecular structure are still poorly understood due to the complexity of the reaction pathways and ultrafast timescales. In order to investigate such light-induced reaction pathways, a new spectroscopic tool has been applied, which combines UV-vis and resonance Raman spectroscopy at multiple excitation wavelengths with electrochemistry in a thin-layer electrochemical cell to study [RuII(tbtpy)2]2+ (tbtpy = tri-tert-butyl-2,2′:6′,2′′-terpyridine) as a model compound for the photo-activated electron donor in structurally related molecular and supramolecular assemblies. The new spectroscopic method substantiates previous suggestions regarding the reduction mechanism of this complex by localizing photo-electrons and identifying structural changes of metastable intermediates along the reaction cascade. This has been realized by monitoring selective enhancement of Raman-active vibrations associated with structural changes upon electronic absorption when tuning the excitation wavelength into new UV-vis absorption bands of intermediate structures. Additional interpretation of shifts in Raman band positions upon reduction with the help of quantum chemical calculations provides a consistent picture of the sequential reduction of the individual terpyridine ligands, i.e., the first reduction results in the monocation [(tbtpy)Ru(tbtpy•)]+, while the second reduction generates [(tbtpy•)Ru(tbtpy•)]0 of triplet multiplicity. Therefore, the combination of this versatile spectro-electrochemical tool allows us to deepen the fundamental understanding of light-induced charge transfer processes in more relevant and complex systems.
- ItemMicro-fluorescene lifetime and spectral imaging of ytterbium doped laser materials(Washington D.C. : Optical Society of America, 2018) Schreiber, T.; Kuhn, S.; Feldkamp, G.; Schuster, Kay; Hein, S.; Eberhardt, Ramona; Tünnermann, AndreasWe present the application of a confocal fluorescence microscope to the analysis of Yb-doped solid-state laser materials, with examples of Yb-doped crystals, photonic crystal fibers and fiber preforms made with different manufacturing processes. Beside the fluorescence lifetime image itself, a microscopic spectral fluorescence emission analysis is presented and spatially resolved emission cross sections are obtained. Doping concentration and its distributions and other laser optical parameters are measured, which help to analyze manufacturing steps. Further properties like photodarkening and saturation are addressed.
- ItemNoise Sources and Requirements for Confocal Raman Spectrometers in Biosensor Applications(Basel : MDPI, 2021) Jahn, Izabella J.; Grjasnow, Alexej; John, Henry; Weber, Karina; Popp, Jürgen; Hauswald, WalterRaman spectroscopy probes the biochemical composition of samples in a non-destructive, non-invasive and label-free fashion yielding specific information on a molecular level. Nevertheless, the Raman effect is very weak. The detection of all inelastically scattered photons with highest efficiency is therefore crucial as well as the identification of all noise sources present in the system. Here we provide a study for performance comparison and assessment of different spectrometers for confocal Raman spectroscopy in biosensor applications. A low-cost, home-built Raman spectrometer with a complementary metal-oxide-semiconductor (CMOS) camera, a middle price-class mini charge-coupled device (CCD) Raman spectrometer and a laboratory grade confocal Raman system with a deeply cooled CCD detector are compared. It is often overlooked that the sample itself is the most important “optical” component in a Raman spectrometer and its properties contribute most significantly to the signal-to-noise ratio. For this purpose, different representative samples: a crystalline silicon wafer, a polypropylene sample and E. coli bacteria were measured under similar conditions using the three confocal Raman spectrometers. We show that biosensor applications do not in every case profit from the most expensive equipment. Finally, a small Raman database of three different bacteria species is set up with the middle price-class mini CCD Raman spectrometer in order to demonstrate the potential of a compact setup for pathogen discrimination.