X-ray mirrors with sub-nanometre figure errors obtained by differential deposition of thin WSi2 films.

Differential deposition by DC magnetron sputtering was applied to correct for figure errors of X-ray mirrors to be deployed on low-emittance synchrotron beamlines. During the deposition process, the mirrors were moved in front of a beam-defining aperture and the required velocity profile was calculated using a deconvolution algorithm. The surface figure was characterized using conventional off-line visible-light metrology instrumentation (long trace profiler and Fizeau interferometer) before and after the deposition. WSi2 was revealed to be a promising candidate material since it conserves the initial substrate surface roughness and limits the film stress to acceptable levels. On a 300 mm-long flat Si mirror the average height errors were reduced by a factor of 20 down to 0.2 nm root mean square. This result shows the suitability of WSi2 for differential deposition. Potential promising applications include the upgrade of affordable, average-quality substrates to the standards of modern synchrotron beamlines.

ID23-2: an automated and high-performance microfocus beamline for macromolecular crystallography at the ESRF.

ID23-2 is a fixed-energy (14.2 keV) microfocus beamline at the European Synchrotron Radiation Facility (ESRF) dedicated to macromolecular crystallography. The optics and sample environment have recently been redesigned and rebuilt to take full advantage of the upgrade of the ESRF to the fourth generation Extremely Brilliant Source (ESRF-EBS). The upgraded beamline now makes use of two sets of compound refractive lenses and multilayer mirrors to obtain a highly intense (>1013 photons s-1) focused microbeam (minimum size 1.5 µm × 3 µm full width at half-maximum). The sample environment now includes a FLEX-HCD sample changer/storage system, as well as a state-of-the-art MD3Up high-precision multi-axis diffractometer. Automatic data reduction and analysis are also provided for more advanced protocols such as synchrotron serial crystallographic experiments.

ID23-2: an automated and high-performance microfocus beamline for macromolecular crystallography at the ESRF. Corrigendum.

A revised version of Table 2 of Nanao et al. [J. Synchrotron Rad. (2022). 29, 581-590] is provided.

X-ray mirror figure correction by differential deposition and off-line metrology.

The surface figure error of a hard X-ray mirror was improved by combining differential deposition and off-line metrology tools. Thin Cr layers were deposited on flat substrates by DC magnetron sputtering. The substrates were moved in front of a beam-defining aperture. The required velocity profile was calculated using a deconvolution algorithm. The Cr thickness profiles were measured directly using hard X-ray reflectivity data. The surface figure was characterized using conventional visible-light metrology instrumentation (long trace profiler) before and after the deposition. The method converges quickly, and after two iterations the mirror surface figure had improved by a factor of 7. The surface roughness evolves with increasing Cr thickness and deteriorates the quality of subsequent multilayer coatings. The mirror curvature can change upon coating, which complicates the interpretation of the surface metrology data. In this context, the role of layer stress is discussed. Potential improvements of the process are also proposed.

Two-dimensional stitching interferometry for self-calibration of high-order additive systematic errors.

Stitching interferometry is performed by collecting interferometric data from overlapped sub-apertures and stitching these data together to provide a full surface map. The propagation of the systematic error in the measured subset data is one of the main error sources in stitching interferometry for accurate reconstruction of the surface topography. In this work, we propose, using the redundancy of the captured subset data, two types of two-dimensional (2D) self-calibration stitching algorithms to overcome this issue by in situ estimating the repeatable high-order additive systematic errors, especially for the application of measuring X-ray mirrors. The first algorithm, called CS short for "Calibrate, and then Stitch", calibrates the high-order terms of the reference by minimizing the de-tilted discrepancies of the overlapped subsets and then stitches the reference-subtracted subsets. The second algorithm, called SC short for "Stitch, and then Calibrate", stitches a temporarily result and then calibrates the reference from the de-tilted discrepancies of the measured subsets and the temporarily stitched result. In the implementation of 2D scans in x- and y-directions, step randomization is introduced to generate nonuniformly spaced subsets which can diminish the periodic stitching errors commonly observed in evenly spaced subsets. The regularization on low-order terms enables a highly flexible option to add the curvature and twist acquired by another system. Both numerical simulations and experiments are carried out to verify the proposed method. All the results indicate that 2D high-order repeatable additive systematic errors can be retrieved from the 2D redundant overlapped data in stitching interferometry.

DABAM: an open-source database of X-ray mirrors metrology.

An open-source database containing metrology data for X-ray mirrors is presented. It makes available metrology data (mirror heights and slopes profiles) that can be used with simulation tools for calculating the effects of optical surface errors in the performances of an optical instrument, such as a synchrotron beamline. A typical case is the degradation of the intensity profile at the focal position in a beamline due to mirror surface errors. This database for metrology (DABAM) aims to provide to the users of simulation tools the data of real mirrors. The data included in the database are described in this paper, with details of how the mirror parameters are stored. An accompanying software is provided to allow simple access and processing of these data, calculate the most usual statistical parameters, and also include the option of creating input files for most used simulation codes. Some optics simulations are presented and discussed to illustrate the real use of the profiles from the database.

Fabrication of ultrashort sub-meter-radius x-ray mirrors using dynamic stencil deposition with figure correction.

This paper presents nanometer-scale production and metrology methods for elliptic-cylindrical x-ray mirrors with an unprecedentedly small tangential radius of curvature of 160 mm. Sub-millimeter-scale figure correction is conducted based on dynamic stencil deposition. The deposition flux through one or two shadow masks is examined by a comparison to a simple model. The masked deposition flux distribution is improved, leading to film thickness profiles that are 50 times sharper in terms of aspect ratio than those obtained using existing differential deposition approaches. Surface roughness deterioration is also effectively suppressed. A 2-mm-long 160-mm-radius mirror is produced with a width of 10 mm and measured using simple interferometry. The results are confirmed by conventional mirror metrology, contact profilometry, and x-ray ptychography. The x-ray focusing profile is diffraction-limited with a 142-nm focus size at a photon energy of 300 eV. The proposed methods have the potential to enhance the ultraprecise fabrication of highly curved mirrors, thus benefiting nanoscale photon-hungry x-ray techniques.

Measurement techniques to improve the accuracy of x-ray mirror metrology using stitching Shack-Hartmann wavefront sensors.

We describe the development of specific measurement protocols to improve the accuracy of surface metrology of x-ray mirrors using a dedicated commercial instrument based on wavefront sensing techniques. This instrument, SHARPeR, uses measurements from a Shack-Hartmann wavefront sensor combined with a sub-aperture stitching method to provide two-dimensional maps of the surface slope errors and can measure curved mirrors above 1 m radii. In this paper, we describe the results of measurement methods developed on a SHARPeR system installed at the European Synchrotron (ESRF) to reduce the contribution of systematic errors to measurements of strongly curved spherical and aspherical x-ray mirrors with intrinsic slope errors of the order of 100-200 nrad rms. We demonstrate how this commercial integrated instrument can provide measurements of these mirrors with comparable accuracy to those measured with a long trace profiler.

Electron microscopy reveals the presence of viruses in mouse embryonic fibroblasts but neither in human embryonic fibroblasts nor in human mesenchymal cells used for hESC maintenance: toward an implementation of microbiological quality assurance program in stem cell banks.

Human embryonic stem cells (hESCs) are expected to open up new avenues in regenerative medicine by allowing the generation of transplantable cells to be used in future cell replacement therapies. Maintenance of hESCs in the presence of xenogenic compounds is likely to prevent their use in future therapeutic applications in humans. Recently, it has been claimed that human foreskin-derived human embryonic fibroblast (HEFs) and human adult marrow cells have the ability to support prolonged expansion of hESCs in culture similar to murine feeders. Here, to minimize the use of xenogenic components for hESC maintenance, we performed transmission electron microscopy-based microbiological studies in an attempt to implement a microbiological Quality Assurance Program in Stem Cell Banks by determining the potential presence of viral particles in MEFs compared with human HEFs and bone marrow-derived mesenchymal cells. We observed in three out of nine MEF samples (33.3%) viruses belonging to the Retroviridae family. Within the Retroviridae family, these viruses have a C morphology, which indicates they belong to the subfamily Orthoretroviridae. In contrast, no viral particles could be observed in either the HEF samples (n = 5) or the human BM-derived mesenchymal cells (n = 9) analyzed. Based on these experimental microbiological data, we recommend the implementation of microbiological Quality Assurance Programs by means of transmission electron microscopy as a routine technique to assess the potential presence of viral particles in any feeder cell used in stem cell banks and support the use of human cells rather than murine cells as feeders to maintain hESC cultures in an undifferentiated state.

In vitro culture of Borrelia garinii results in loss of flagella and decreased invasiveness.

A virulent, low-passage culture of a tick-derived strain of Borrelia garinii was subjected to serial in vitro passages, from which inoculations were made into C3H/HeN mice. A full display of pathogenicity was observed through passage 4, as measured by cultures of ear punch biopsy samples and internal organs and determination of tibiotarsal joint swelling. Decreased dissemination through skin and infection of internal organs were observed beginning at passage 6. These losses correlated with both the selection of clones harboring 21% less flagella than the parent strain, as seen by electron microscopy, and loss of the motility of the higher passages, as demonstrated by a swarm assay. However, during the chronic phase (3 months after infection), spirochetes were cultured from the bladder and kidney of a mouse inoculated with passage 12. The kidney isolate had the same number of flagella and motility as the original low-passage isolate. Although we can't exclude the possibility that other subtle variations may be arising given the uncloned nature of the isolate, we have found a strong association between loss of flagella and decreased invasiveness. Arthritogenicity progressively decreased with passages, so that only 12.5% of chronically infected mice inoculated with passage 29 still presented with joint swelling, concurrent with a decrease in the staining intensity in a Southern blot with a vlsE-based probe. These results suggest a multifactorial model in which the number of flagella drives the invasiveness of this agent, while plasmid-associated factors are responsible for triggering arthritogenicity.