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Protein interaction and protein imaging strongly benefit from the advancements in time-resolved and superresolution fluorescence microscopic techniques. However, the techniques were typically applied separately and ex vivo because of technical challenges and the absence of suitable fluorescent protein pairs. Here, we show correlative in vivo fluorescence lifetime imaging microscopy Förster resonance energy transfer (FLIM-FRET) and stimulated emission depletion (STED) microscopy to unravel protein mechanics and structure in living cells. We use magnetotactic bacteria as a model system where two proteins, MamJ and MamK, are used to assemble magnetic particles called magnetosomes. The filament polymerizes out of MamK and the magnetosomes are connected via the linker MamJ. Our system reveals that bacterial filamentous structures are more fragile than the connection of biomineralized particles to this filament. More importantly, we anticipate the technique to find wide applicability for the study and quantification of biological processes in living cells and at high resolution.
Quantum dots increasingly gain popularity for in vivo applications. However, their delivery and accumulation into cells can be challenging and there is still lack of detailed information. Thereby, the application of advanced fluorescence techniques can expand the portfolio of useful parameters for a more comprehensive evaluation. Here, we encapsulated hydrophilic quantum dots into liposomes for studying cellular uptake of these so-called lipodots into living cells. First, we investigated photophysical properties of free quantum dots and lipodots observing changes in the fluorescence decay time and translational diffusion behaviour. In comparison to empty liposomes, lipodots exhibited an altered zeta potential, whereas their hydrodynamic size did not change. Fluorescence lifetime imaging microscopy (FLIM) and fluorescence correlation spectroscopy (FCS), both combined with two-photon excitation (2P), were used to investigate the interaction behaviour of lipodots with an insect epithelial tissue. In contrast to the application of free quantum dots, their successful delivery into the cytosol of salivary gland duct cells could be observed when applying lipodots. Lipodots with different lipid compositions and surface charges did not result in considerable differences in the intracellular labelling pattern, luminescence decay time and diffusion behaviour. However, quantum dot degradation after intracellular accumulation could be assumed from reduced luminescence decay times and blue-shifted luminescence signals. In addition to single diffusing quantum dots, possible intracellular clustering of quantum dots could be assumed from increased diffusion times. Thus, by using a simple and manageable liposome carrier system, 2P-FLIM and 2P-FCS recording protocols could be tested, which are promising for investigating the fate of quantum dots during cellular interaction.
Autocrats utilize (nominally) democratic elections, to claim procedural legitimation. To secure their political survival in these elections, they have an extensive menu of manipulation at their disposal. These manipulations are not only addressed at contestation but also inclusiveness of the elections. Although autocrats formally claim universal suffrage, informal restrictions and practices are implemented. Analyzing elections held between 1970 and 2020 in electoral autocracies, I find empiric evidence for strategic adjustments of suffrage rights as a response to electoral contexts.
"Bei der Entwicklung von Informationssystemen, die eine durchgängige Unterstützung aller in einem Unternehmen zu bewältigenden Abläufe bieten können, ist nicht in erster Linie von den Leistungsmerkmalen konkreter EDV-Systeme auszugehen. Vielmehr sind hierbei die strategischen Unternehmensziele sowie die für die Erfüllung dieser Ziele erforderlichen Vorgangsketten zugrunde zu legen. Die EDV-Technik dient dann lediglich der Umsetzung der unternehmerischen Zielsetzung mit Hilfe konkreter Informationssyssteme und sollte die freie Gestaltung von Arbeitsabläufen und Organisationskonzepten ermöglichen."[1]
Die Suche nach sicheren Konzepten, angepaßt an individuelle Unternehmensstrukturen, ist an unterschiedlichen Forschungseinrichtungen ein Thema. An der Nahtstelle zwischen den Fachbereichen Betriebswirtschaft und Maschinenbau ist dies auch an der Technischen Fachhochschule Wildau ein Thema.
The article investigates the correlation between the factors of enhancing traffic safety on highways and parameters of the economic growth in Russia and in countries with transition economy; such correlation does not always lead to traffic safety enhancement. As a rule, the population motorization (car density) level growth in such countries is not accompanied by efficient actions on accident rate decrease. Among the factors enhancing traffic safety on highways are the efficiency of traffic infrastructure planning (changing and removing traffic flows from cities’ centers, creation of a large amount of pedestrian and bicycle lanes separated from traffic flows) as well as harmonization of physical infrastructure and safety goals, ensuring conjugation of the developed transport infrastructure with the decrease of the share of private vehicle fleet and increase of the share of public transport.
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