25 research outputs found

    Characterization of Memory and Measurement History in Photoconductivity of Nanocyrstal Arrays

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    Photoconductivity in nanocrystal films has been previously characterized, but memory effects have received little attention despite their importance for device applications. We show that the magnitude and temperature dependence of the photocurrent in CdSe/ZnS core-shell nanocrystal arrays depends on the illumination and electric field history. Changes in photoconductivity occur on a few-hour timescale, and subband gap illumination of nanocrystals prior to measurements modifies the photocurrent more than band gap illumination. The observed effects can be explained by charge traps within the band gap that are filled or emptied, which may alter nonradiative recombination processes and affect photocurrent

    Fabrication of Nano-Scale Gaps in Integrated Circuits

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    Nano-size objects like metal clusters present an ideal system for the study of quantum phenomena and for constructing practical quantum devices. Integrating these small objects in a macroscopic circuit is, however, a difficult task. So far the nanoparticles have been contacted and addressed by highly sophisticated techniques which are not suitable for large scale integration in macroscopic circuits. We present an optical lithography method that allows for the fabrication of a network of electrodes which are separated by gaps of controlled nanometer size. The main idea is to control the gap size with subnanometer precision using a structure grown by molecular beam epitaxy.Comment: 4 pages, 3 figure

    Resolving ESCRT-III Spirals at the Intercellular Bridge of Dividing Cells Using 3D STORM

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    Summary: The ESCRT machinery mediates membrane fission in a variety of processes in cells. According to current models, ESCRT-III proteins drive membrane fission by assembling into helical filaments on membranes. Here, we used 3D STORM imaging of endogenous ESCRT-III component IST1 to reveal the evolution of the structural organization of ESCRT-III in mammalian cytokinetic abscission. Using this approach, ESCRT-III ring and spiral assemblies were resolved and characterized at different stages of abscission. Visualization of IST1 structures in cells lacking the microtubule-severing enzyme spastin and in cells depleted of specific ESCRT-III components or the ATPase VPS4 demonstrated the contribution of these components to the organization and function of ESCRTs in cells. This work provides direct evidence that ESCRT-III proteins form helical filaments to mediate their function in cells and raises new mechanistic scenarios for ESCRT-driven cytokinetic abscission. : The ESCRT complex drives membrane constriction and fission in a variety of process in cells. In this work, Goliand et al. used 3D STORM imaging to resolve the structure of the ESCRT-III complex during abscission of the intercellular bridge connecting two dividing cells, highlighting mechanistic steps in ESCRT cellular function. Keywords: super resolution microscopy, cytokinesis, abscission, ESCRT machinery, membrane fissio

    Interpreting the Energy-Dependent Anisotropy of Colloidal Nanorods Using Ensemble and Single-Particle Spectroscopy

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    We report the use of polarized excitation spectroscopy in the study of the polarized optical properties of II–VI semiconducting nanorods. This technique provides a quantitative measure for analyzing the polarization-dependent electronic structure of ensembles of semiconducting nanoparticles in colloidal solutions. We develop a procedure to quantify the anisotropy of nanorod excitations and the influence of the dielectric environment which yields results in qualitative agreement with theoretical predictions of nanorod absorption properties. Excitation measurements of nanorod ensembles and single-particle excitation and emission polarization measurements are used to interpret ensemble measurements on the molecular frame. At the single-particle level, we find a large dispersion in the angles between excitation and emission polarization curves. This contrasts with the conventional picture of CdSe electronic transitions, in which the crystal axes dictate the symmetry of excitations. Such geometrically heterogeneous excitations suggest that perturbations of the physical structure adjust the polarization properties of individual nanorods from those of an ideal theoretical nanorod

    Colocalization and Disposition of Cellulosomes in Clostridium clariflavum as Revealed by Correlative Superresolution Imaging

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    Cellulosomes are multienzyme complexes produced by anaerobic, cellulolytic bacteria for highly efficient breakdown of plant cell wall polysaccharides. Clostridium clariflavum is an anaerobic, thermophilic bacterium that produces the largest assembled cellulosome complex in nature to date, comprising three types of scaffoldins: a primary scaffoldin, ScaA; an adaptor scaffoldin, ScaB; and a cell surface anchoring scaffoldin, ScaC. This complex can contain 160 polysaccharide-degrading enzymes. In previous studies, we proposed potential types of cellulosome assemblies in C. clariflavum and demonstrated that these complexes are released into the extracellular medium. In the present study, we explored the disposition of the highly structured, four-tiered cell-anchored cellulosome complex of this bacterium. Four separate, integral cellulosome components were subjected to immunolabeling: ScaA, ScaB, ScaC, and the cellulosome’s most prominent enzyme, GH48. Imaging of the cells by correlating scanning electron microscopy and three-dimensional (3D) superresolution fluorescence microscopy revealed that some of the protuberance-like structures on the cell surface represent cellulosomes and that the components are highly colocalized and organized by a defined hierarchy on the cell surface. The display of the cellulosome on the cell surface was found to differ between cells grown on soluble or insoluble substrates. Cell growth on microcrystalline cellulose and wheat straw exhibited dramatic enhancement in the amount of cellulosomes displayed on the bacterial cell surface

    Phospholipid imbalance impairs autophagosome completion

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    Autophagy, a conserved eukaryotic intracellular catabolic pathway, maintains cell homeostasis by lysosomal degradation of cytosolic material engulfed in double membrane vesicles termed autophagosomes, which form upon sealing of single-membrane cisternae called phagophores. While the role of phosphatidylinositol 3-phosphate (PI3P) and phosphatidylethanolamine (PE) in autophagosome biogenesis is well-studied, the roles of other phospholipids in autophagy remain rather obscure. Here we utilized budding yeast to study the contribution of phosphatidylcholine (PC) to autophagy. We reveal for the first time that genetic loss of PC biosynthesis via the CDP-DAG pathway leads to changes in lipid composition of autophagic membranes, specifically replacement of PC by phosphatidylserine (PS). This impairs closure of the autophagic membrane and autophagic flux. Consequently, we show that choline-dependent recovery of de novo PC biosynthesis via the CDP-choline pathway restores autophagosome formation and autophagic flux in PC-deficient cells. Our findings therefore implicate phospholipid metabolism in autophagosome biogenesis

    Deposition of collagen type I onto skeletal endothelium reveals a new role for blood vessels in regulating bone morphology

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    \u3cp\u3eRecently, blood vessels have been implicated in the morphogenesis of various organs. The vasculature is also known to be essential for endochondral bone development, yet the underlying mechanism has remained elusive. We show that a unique composition of blood vessels facilitates the role of the endothelium in bone mineralization and morphogenesis. Immunostaining and electron microscopy showed that the endothelium in developing bones lacks basement membrane, which normally isolates the blood vessel from its surroundings. Further analysis revealed the presence of collagen type I on the endothelial wall of these vessels. Because collagen type I is the main component of the osteoid, we hypothesized that the bone vasculature guides the formation of the collagenous template and consequently of the mature bone. Indeed, some of the bone vessels were found to undergo mineralization. Moreover, the vascular pattern at each embryonic stage prefigured the mineral distribution pattern observed one day later. Finally, perturbation of vascular patterning by overexpressing Vegf in osteoblasts resulted in abnormal bone morphology, supporting a role for blood vessels in bone morphogenesis. These data reveal the unique composition of the endothelium in developing bones and indicate that vascular patterning plays a role in determining bone shape by forming a template for deposition of bone matrix.\u3c/p\u3
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