No abstract

Hard x-ray microscopy with nanometer resolution will open frontiers in the study of materials and devices, environmental sciences, and life sciences by utilizing the unique characterization capabilities of x-rays. Here we report two-dimensional nanofocusing by multilayer Laue lenses (MLLs), a type of diffractive optics that is in principle capable of focusing x-rays to 1 nm. We demonstrate focusing to a 25 × 27 nm(2) FWHM spot with an efficiency of 2% at a photon energy of 12 keV, and to a 25 × 40 nm(2) FWHM spot with an efficiency of 17% at a photon energy of 19.5 keV.

We have used coherent X-ray diffraction experiments to characterize both the 1-D and 2-D foci produced by nanofocusing Kirkpatrick-Baez (K-B) mirrors, and we find agreement. Algorithms related to ptychography were used to obtain a 3-D reconstruction of a focused hard X-ray beam waist, using data measured when the mirrors were not optimally aligned. Considerable astigmatism was evident in the reconstructed complex wavefield. Comparing the reconstructed wavefield for a single mirror with a geometrical projection of the wavefront errors expected from optical metrology data allowed us to diagnose a 40 μrad misalignment in the incident angle of the first mirror, which had occurred during the experiment. Good agreement between the reconstructed wavefront obtained from the X-ray data and off-line metrology data obtained with visible light demonstrates the usefulness of the technique as a metrology and alignment tool for nanofocusing X-ray optics.

We describe progress in the fabrication of short-focal-length total-external-reflection Kirkpatrick-Baez x-ray mirrors with ultralow figure errors. The short focal length optics produce nanoscale beams (<100nm) on conventional (∼64m long) beamlines at third generation synchrotron sources. The total-external reflection optics are inherently achromatic and efficiently focus a white (polychromatic) or a tunable monochromatic spectrum of x rays. The ability to focus independent of wavelength allows novel new experimental capabilities. Mirrors have been fabricated both by computer assisted profiling (differential polishing) and by profile coating (coating through a mask onto ultra-smooth surfaces). A doubly focused 85×95nm2 hard x-ray nanobeam has been obtained on the UNICAT beamline 34-ID at the Advanced Photon Source. The performance of the mirrors, techniques for characterizing the spot size, and factors limiting focusing performance are discussed.

RFLAT-1/KLF13, a member of the Krü ppel-like family of transcription factors, was identified as a transcription factor expressed 3-5 days after T lymphocyte activation. It binds to the promoter of the chemokine gene RANTES (regulated on activation normal T cell expressed and secreted) and regulates its ''late'' expression in activated T-cells. In this study, a series of experiments to define the functional domains of RFLAT-1/KLF13 were undertaken to further advance the understanding of the molecular mechanisms underlying transcriptional regulation by this factor. Using the GAL4 fusion system, distinct transcriptional activation and repression domains were identified. The RFLAT-1 minimum activation domain is localized to amino acids 1-35, whereas the repression domain resides in amino acids 67-168. Deletion analysis on the RFLAT-1 protein further supports these domain functions. The RFLAT-1 activation domain is similar to that of its closest family member, basic transcription element-binding protein 1. This domain is highly hydrophobic, and site-directed mutagenesis demonstrated that both negatively charged and hydrophobic residues are important for transactivation. The nuclear localization signal of RFLAT-1 was also identified using the RFLAT-1/green fluorescence protein fusion approach. RFLAT-1 contains two potent, independent nuclear localization signals; one is immediately upstream of the zinc finger DNA-binding domain, and the other is within the zinc fingers. Using mutational analysis, we also determined that the critical binding sequence of RFLAT-1 is CTCCC. The intact CTCCC box on the RANTES promoter is necessary for RFLAT-1-mediated RANTES transcription and is also required for the synergy between RFLAT-1 and NF-B proteins.