Vision and refraction in Australian preschool children : measures, prevalence, and associated factors

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Whole-Cell Analysis of Low-Density Lipoprotein Uptake by Macrophages Using STEM Tomography

Nanoparticles of heavy materials such as gold can be used as markers in quantitative electron microscopic studies of protein distributions in cells with nanometer spatial resolution. Studying nanoparticles within the context of cells is also relevant for nanotoxicological research. Here, we report a method to quantify the locations and the number of nanoparticles, and of clusters of nanoparticles inside whole eukaryotic cells in three dimensions using scanning transmission electron microscopy (STEM) tomography. Whole-mount fixed cellular samples were prepared, avoiding sectioning or slicing. The level of membrane staining was kept much lower than is common practice in transmission electron microscopy (TEM), such that the nanoparticles could be detected throughout the entire cellular thickness. Tilt-series were recorded with a limited tilt-range of 80° thereby preventing excessive beam broadening occurring at higher tilt angles. The 3D locations of the nanoparticles were nevertheless determined with high precision using computation. The obtained information differed from that obtained with conventional TEM tomography data since the nanoparticles were highlighted while only faint contrast was obtained on the cellular material. Similar as in fluorescence microscopy, a particular set of labels can be studied. This method was applied to study the fate of sequentially up-taken low-density lipoprotein (LDL) conjugated to gold nanoparticles in macrophages. Analysis of a 3D reconstruction revealed that newly up-taken LDL-gold was delivered to lysosomes containing previously up-taken LDL-gold thereby forming onion-like clusters

Whole-cell analysis of low-density lipoprotein uptake by macrophages using STEM tomography

Nanoparticles of heavy materials such as gold can be used as markers in quantitative electron microscopic studies of protein distributions in cells with nanometer spatial resolution. Studying nanoparticles within the context of cells is also relevant for nanotoxicological research. Here, we report a method to quantify the locations and the number of nanoparticles, and of clusters of nanoparticles inside whole eukaryotic cells in three dimensions using scanning transmission electron microscopy (STEM) tomography. Whole-mount fixed cellular samples were prepared, avoiding sectioning or slicing. The level of membrane staining was kept much lower than is common practice in transmission electron microscopy (TEM), such that the nanoparticles could be detected throughout the entire cellular thickness. Tilt-series were recorded with a limited tilt-range of 80u thereby preventing excessive beam broadening occurring at higher tilt angles. The 3D locations of the nanoparticles were nevertheless determined with high precision using computation. The obtained information differed from that obtained with conventional TEM tomography data since the nanoparticles were highlighted while only faint contrast was obtained on the cellular material. Similar as in fluorescence microscopy, a particular set of labels can be studied. This method was applied to study the fate of sequentially up-taken low-density lipoprotein (LDL) conjugated to gold nanoparticles in macrophages. Analysis of a 3D reconstruction revealed that newly up-taken LDL-gold was delivered to lysosomes containing previously up-taken LDL-gold thereby forming onion-like clusters

Electron microscopy of nanoparticle superlattice formation at a solid-liquid interface in nonpolar liquids

Nanoparticle superlattice films form at the solid-liquid interface and are important for mesoscale materials, but are notoriously difficult to analyze before they are fully dried. Here, the early stages of nanoparticle assembly were studied at solid-liquid interfaces using liquid-phase electron microscopy. Oleylamine-stabilized gold nanoparticles spontaneously formed thin layers on a silicon nitride (SiN) membrane window of the liquid enclosure. Dense packings of hexagonal symmetry were obtained for the first monolayer independent of the nonpolar solvent type. The second layer, however, exhibited geometries ranging from dense packing in a hexagonal honeycomb structure to quasi-crystalline particle arrangements depending on the dielectric constant of the liquid. The complex structures formed by the weaker interactions in the second particle layer were preserved, while the surface remained immersed in liquid. Fine-tuning the properties of the involved materials can thus be used to control the three-dimensional geometry of a superlattice including quasi-crystals

Exploring neuropsychological effects of a self-monitoring intervention for ADHD-symptoms in school

Children who have symptoms of Attention Deficit/Hyperactivity Disorder (ADHD) often experience disruptive and off-task behavior and lower school performance than would be expected based on their cognitive abilities. These behavior and achievement difficulties are a challenge to teachers, who often provide inclusive education for children with ADHD. This study explored whether a self-monitoring intervention can help children with ADHD-symptoms to reduce off-task behavior as well as improve their cognition. The participating children were seven boys in special needs education with an age between nine and twelve. The students used an interval timer to remind themselves to monitor whether they were still on task. During math classes, observations, teacher ratings and neuropsychological tests were assessed. The results showed that off-task behavior was significantly reduced during the period the interval timer was used compared to baseline (reduction from 46.8 to 27.3%), as measured by observations (effect size: η2p = .83) and this was confirmed by teacher ratings (effect size: η2p = .69). With respect to cognition, children only showed significant improvements in inhibition (effect sizes: Cohen's d = 2.62 and 1.24). The teachers as well as students evaluated the intervention mainly as positive. In line with previous studies, we found that that a self-monitoring intervention can be beneficial for children with ADHD-symptoms. Larger studies including a control group and blind observers are necessary to establish these results and to investigate the underlying mechanisms

Measuring and managing changes in estuaries and lagoons: Morphological and eco-toxicological aspects

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