Soft X-ray microcopy has resolved 30 nm structures in biological cells. To protect the cells from radiation damage caused by X-rays, imaging of the samples has to be performed at cryogenic temperatures, which makes it possible to take multiple images of a single cell. Due to the small numerical aperture of zone plates, X-ray objectives have a depth of focus on the order of several microns. By treating the X-ray microscopic images as projections of the sample absorption, computed tomography (CT) can be performed. Since cryogenic biological samples are resistant to radiation damage, it is possible to reconstruct frozen-hydrated cells imaged with a full-field X-ray microscope. This approach is used to obtain three-dimensional information about the location of specific proteins in cells. To localize proteins in cells, immunolabelling with strongly X-ray absorbing nanoparticles was performed. With the new tomography setup developed for the X-ray microscope XM-1 installed at the ALS, we have performed tomography of immunolabelled frozen-hydrated cells to detect protein distributions inside of cells. As a first example, the distribution of the nuclear protein, male specific lethal 1 (MSL-1) in the Drosophila melanogaster cell was studied.
High-resolution x-ray microscopy makes use of nanostructured diffractive optics such as Fresnel zone plates made of nickel. These micro zone plates are manufactured using microscopic galvanoforms and electrodeposition techniques. Copolymer galvanoforms for nickel micro zone plates were irradiated with high doses of x-ray radiation to increase the degree of cross-linking of the copolymer network; in this way, exact pattern replication in the galvanoform was obtained for zone aspect ratios of up to 8:1. Using these galvanoforms, nickel micro zone plates were produced for the soft x-ray wavelength λ=2.4 nm, with smallest zone widths of 30 nm and 40 nm, achieving first-order diffraction efficiencies of up to 13% and 20%, respectively.
The inner surface of the ring vacuum chambers of the US Spallation Neutron Source (SNS) will be coated with ~100 nm of Titanium Nitride (TiN). This is to minimize the secondary electron yield (SEY) from the chamber wall, and thus avoid the so-called e-p instability caused by electron multipacting as observed in a few high-intensity proton storage rings. Both DC sputtering and DCmagnetron sputtering were conducted in a test chamber of relevant geometry to SNS ring vacuum chambers. Auger Electron Spectroscopy (AES) and Rutherford Back Scattering (RBS) were used to analyze the coatings for thickness, stoichiometry and impurity. Excellent results were obtained with magnetron sputtering. The development of the parameters for the coating process and the surface analysis results are presented.
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