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- ItemEpidermal growth factor receptor subunit locations determined in hydrated cells with environmental scanning electron microscopy(London : Nature Publishing Group, 2013) Peckys, Diana B; Baudoin, Jean-Pierre; Eder, Magdalena; Werner, Ulf; de Jonge, NielsImaging single epidermal growth factor receptors (EGFR) in intact cells is presently limited by the available microscopy methods. Environmental scanning electron microscopy (ESEM) of whole cells in hydrated state in combination with specific labeling with gold nanoparticles was used to localize activated EGFRs in the plasma membranes of COS7 and A549 cells. The use of a scanning transmission electron microscopy (STEM) detector yielded a spatial resolution of 3 nm, sufficient to identify the locations of individual EGFR dimer subunits. The sizes and distribution of dimers and higher order clusters of EGFRs were determined. The distance between labels bound to dimers amounted to 19 nm, consistent with a molecular model. A fraction of the EGFRs was found in higher order clusters with sizes ranging from 32–56 nm. ESEM can be used for quantitative whole cell screening studies of membrane receptors, and for the study of nanoparticle-cell interactions in general.
- ItemSize control in mammalian cells involves modulation of both growth rate and cell cycle duration(London : Nature Publishing Group, 2018) Cadar, Clotilde; Monnier, Sylvain; Grilli, Jacopo; Sáez, Pablo J.; Srivastava, Nishit; Attia, Rafaele; Terriac, Emmanuel; Baum, Buzz; Cosentino-Lagomarsino, Marco; Piel, MatthieuDespite decades of research, how mammalian cell size is controlled remains unclear because of the difficulty of directly measuring growth at the single-cell level. Here we report direct measurements of single-cell volumes over entire cell cycles on various mammalian cell lines and primary human cells. We find that, in a majority of cell types, the volume added across the cell cycle shows little or no correlation to cell birth size, a homeostatic behavior called “adder”. This behavior involves modulation of G1 or S-G2 duration and modulation of growth rate. The precise combination of these mechanisms depends on the cell type and the growth condition. We have developed a mathematical framework to compare size homeostasis in datasets ranging from bacteria to mammalian cells. This reveals that a near-adder behavior is the most common type of size control and highlights the importance of growth rate modulation to size control in mammalian cells.
- ItemDissection of iron signaling and iron accumulation by overexpression of subgroup Ib bHLH039 protein(London : Nature Publishing Group, 2017) Naranjo-Arcos, Maria Augusta; Maurer, Felix; Meiser, Johannes; Pateyron, Stephanie; Fink-Straube, Claudia; Bauer, PetraIron is an essential growth determinant for plants, and plants acquire this micronutrient in amounts they need in their environment. Plants can increase iron uptake in response to a regulatory transcription factor cascade. Arabidopsis thaliana serves as model plant to identify and characterize iron regulation genes. Here, we show that overexpression of subgroup Ib bHLH transcription factor bHLH039 (39Ox) caused constitutive iron acquisition responses, which resulted in enhanced iron contents in leaves and seeds. Transcriptome analysis demonstrated that 39Ox plants displayed simultaneously gene expression patterns characteristic of iron deficiency and iron stress signaling. Thereby, we could dissect iron deficiency response regulation. The transcription factor FIT, which is required to regulate iron uptake, was essential for the 39Ox phenotype. We provide evidence that subgroup Ib transcription factors are involved in FIT transcriptional regulation. Our findings pose interesting questions to the feedback control of iron homeostasis.
- ItemDynamic fine-tuning of CAR-T cell therapy(London : Nature Publishing Group, 2023) Trehin, Pierre V.M.; Muñoz-Guamuro, Geisler; Weber, Wilfried[No abstract available]
- ItemX-ray imaging with scintillator-sensitized hybrid organic photodetectors(London : Nature Publishing Group, 2015) Büchele, Patric; Richter, Moses; Tedde, Sandro F.; Matt, Gebhard J.; Ankah, Genesis N.; Fischer, Rene; Biele, Markus; Metzger, Wilhelm; Lilliu, Samuele; Bikondoa, Oier; Macdonald, J. Emyr; Brabec, Christoph J.; Kraus, Tobias; Lemmer, Uli; Schmidt, OliverMedical X-ray imaging requires cost-effective and high-resolution flat-panel detectors for the energy range between 20 and 120 keV. Solution-processed photodetectors provide the opportunity to fabricate detectors with a large active area at low cost. Here, we present a disruptive approach that improves the resolution of such detectors by incorporating terbium-doped gadolinium oxysulfide scintillator particles into an organic photodetector matrix. The X-ray induced light emission from the scintillators is absorbed within hundreds of nanometres, which is negligible compared with the pixel size. Hence, optical crosstalk, a limiting factor in the resolution of scintillator-based X-ray detectors, is minimized. The concept is validated with a 256 × 256 pixel detector with a resolution of 4.75 lp mm−1 at a MTF = 0.2, significantly better than previous stacked scintillator-based flat-panel detectors. We achieved a resolution that proves the feasibility of solution-based detectors in medical applications. Time-resolved electrical characterization showed enhanced charge carrier mobility with increased scintillator filling, which is explained by morphological changes.
- ItemOptoregulated force application to cellular receptors using molecular motors(London : Nature Publishing Group, 2021) Zheng, Yijun; Han, Mitchell K.L.; Zhao, Renping; Blass, Johanna; Zhang, Jingnan; Zhou, Dennis W.; Colard-Itté, Jean-Rémy; Dattler, Damien; Çolak, Arzu; Hoth, Markus; García, Andrés J.; Qu, Bin; Bennewitz, Roland; Giuseppone, Nicolas; del Campo, AránzazuProgress in our understanding of mechanotransduction events requires noninvasive methods for the manipulation of forces at molecular scale in physiological environments. Inspired by cellular mechanisms for force application (i.e. motor proteins pulling on cytoskeletal fibers), we present a unique molecular machine that can apply forces at cell-matrix and cell-cell junctions using light as an energy source. The key actuator is a light-driven rotatory molecular motor linked to polymer chains, which is intercalated between a membrane receptor and an engineered biointerface. The light-driven actuation of the molecular motor is converted in mechanical twisting of the entangled polymer chains, which will in turn effectively “pull” on engaged cell membrane receptors (e.g., integrins, T cell receptors) within the illuminated area. Applied forces have physiologically-relevant magnitude and occur at time scales within the relevant ranges for mechanotransduction at cell-friendly exposure conditions, as demonstrated in force-dependent focal adhesion maturation and T cell activation experiments. Our results reveal the potential of nanomotors for the manipulation of living cells at the molecular scale and demonstrate a functionality which at the moment cannot be achieved by other technologies for force application.
- ItemSalt concentration and charging velocity determine ion charge storage mechanism in nanoporous supercapacitors(London : Nature Publishing Group, 2018) Prehal, C.; Koczwara, C.; Amenitsch, H.; Presser, V.; Paris, O.A fundamental understanding of ion charge storage in nanoporous electrodes is essential to improve the performance of supercapacitors or devices for capacitive desalination. Here, we employ in situ X-ray transmission measurements on activated carbon supercapacitors to study ion concentration changes during electrochemical operation. Whereas counter-ion adsorption was found to dominate at small electrolyte salt concentrations and slow cycling speed, ion replacement prevails for high molar concentrations and/or fast cycling. Chronoamperometry measurements reveal two distinct time regimes of ion concentration changes. In the first regime the supercapacitor is charged, and counter- and co-ion concentration changes align with ion replacement and partially co-ion expulsion. In the second regime, the electrode charge remains constant, but the total ion concentration increases. We conclude that the initial fast charge neutralization in nanoporous supercapacitor electrodes leads to a non-equilibrium ion configuration. The subsequent, charge-neutral equilibration slowly increases the total ion concentration towards counter-ion adsorption.
- ItemLipid droplets as a novel cargo of tunnelling nanotubes in endothelial cells(London : Nature Publishing Group, 2015) Astanina, Ksenia; Koch, Marcus; Jüngst, Christian; Zumbusch, Andreas; Kiemer, Alexandra K.Intercellular communication is a fundamental process in the development and functioning of multicellular organisms. Recently, an essentially new type of intercellular communication, based on thin membrane channels between cells, has been reported. These structures, termed intercellular or tunnelling nanotubes (TNTs), permit the direct exchange of various components or signals (e.g., ions, proteins, or organelles) between non-adjacent cells at distances over 100 μm. Our studies revealed the presence of tunnelling nanotubes in microvascular endothelial cells (HMEC-1). The TNTs were studied with live cell imaging, environmental scanning electron microscopy (ESEM), and coherent anti-Stokes Raman scattering spectroscopy (CARS). Tunneling nanotubes showed marked persistence: the TNTs could connect cells over long distances (up to 150 μm) for several hours. Several cellular organelles were present in TNTs, such as lysosomes and mitochondria. Moreover, we could identify lipid droplets as a novel type of cargo in the TNTs. Under angiogenic conditions (VEGF treatment) the number of lipid droplets increased significantly. Arachidonic acid application not only increased the number of lipid droplets but also tripled the extent of TNT formation. Taken together, our results provide the first demonstration of lipid droplets as a cargo of TNTs and thereby open a new field in intercellular communication research.