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    Quantification of silver nanoparticle uptake and distribution within individual human macrophages by FIB/SEM slice and view
    (London : Biomed Central, 2017-3-21) Guehrs, Erik; Schneider, Michael; Günther, Christian M.; Hessing, Piet; Heitz, Karen; Wittke, Doreen; López-Serrano Oliver, Ana; Jakubowski, Norbert; Plendl, Johanna; Eisebitt, Stefan; Haase, Andrea
    Background: Quantification of nanoparticle (NP) uptake in cells or tissues is very important for safety assessment. Often, electron microscopy based approaches are used for this purpose, which allow imaging at very high resolution. However, precise quantification of NP numbers in cells and tissues remains challenging. The aim of this study was to present a novel approach, that combines precise quantification of NPs in individual cells together with high resolution imaging of their intracellular distribution based on focused ion beam/ scanning electron microscopy (FIB/SEM) slice and view approaches. Results: We quantified cellular uptake of 75 nm diameter citrate stabilized silver NPs (Ag 75 Cit) into an individual human macrophage derived from monocytic THP-1 cells using a FIB/SEM slice and view approach. Cells were treated with 10 μg/ml for 24 h. We investigated a single cell and found in total 3138 ± 722 silver NPs inside this cell. Most of the silver NPs were located in large agglomerates, only a few were found in clusters of fewer than five NPs. Furthermore, we cross-checked our results by using inductively coupled plasma mass spectrometry and could confirm the FIB/SEM results. Conclusions: Our approach based on FIB/SEM slice and view is currently the only one that allows the quantification of the absolute dose of silver NPs in individual cells and at the same time to assess their intracellular distribution at high resolution. We therefore propose to use FIB/SEM slice and view to systematically analyse the cellular uptake of various NPs as a function of size, concentration and incubation time.
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    CAMP@FLASH: an end-station for imaging, electron- and ion-spectroscopy, and pump–probe experiments at the FLASH free-electron laser
    (Chester : IUCr, 2018-8-2) Erk, Benjamin; Müller, Jan P.; Bomme, Cédric; Boll, Rebecca; Brenner, Günter; Chapman, Henry N.; Correa, Jonathan; Düsterer, Stefan; Dziarzhytski, Siarhei; Eisebitt, Stefan; Graafsma, Heinz; Grunewald, Sören; Gumprecht, Lars; Hartmann, Robert; Hauser, Günter; Keitel, Barbara; von Korff Schmising, Clemens; Kuhlmann, Marion; Manschwetus, Bastian; Mercadier, Laurent; Müller, Erland; Passow, Christopher; Plönjes, Elke; Ramm, Daniel; Rompotis, Dimitrios; Rudenko, Artem; Rupp, Daniela; Sauppe, Mario; Siewert, Frank; Schlosser, Dieter; Strüder, Lothar; Swiderski, Angad; Techert, Simone; Tiedtke, Kai; Tilp, Thomas; Treusch, Rolf; Schlichting, Ilme; Ullrich, Joachim; Moshammer, Robert; Möller, Thomas; Rolles, Daniel
    The non-monochromatic beamline BL1 at the FLASH free-electron laser facility at DESY was upgraded with new transport and focusing optics, and a new permanent end-station, CAMP, was installed. This multi-purpose instrument is optimized for electron- and ion-spectroscopy, imaging and pump–probe experiments at free-electron lasers. It can be equipped with various electron- and ion-spectrometers, along with large-area single-photon-counting pnCCD X-ray detectors, thus enabling a wide range of experiments from atomic, molecular, and cluster physics to material and energy science, chemistry and biology. Here, an overview of the layout, the beam transport and focusing capabilities, and the experimental possibilities of this new end-station are presented, as well as results from its commissioning.