Stability investigation of a cryo soft x-ray microscope by fiber interferometry.

We present a stability investigation of the Stockholm laboratory cryo soft x-ray microscope. The microscope operates at a wavelength of 2.48 nm and can image biological samples at liquid-nitrogen temperatures in order to mitigate radiation damage. We measured the stability of the two most critical components, sample holder and optics holder, in vacuo and at cryo temperatures at both short and long time scales with a fiber interferometer. Results revealed vibrations in the kHz range, originating mainly from a turbo pump, as well as long term drifts in connection with temperature fluctuations. With improvements in the microscope, earlier stability issues vanished and close-to diffraction-limited imaging could be achieved. Moreover, our investigation shows that fiber interferometers are a powerful tool in order to investigate position-sensitive setups at the nanometer level.

Segway® related injuries in Vienna: report from the Lorenz Böhler Trauma Centre.

INTRODUCTION: The Segway® vehicle facilitates a new way of eco-friendly mobility and is currently used all over the world. In the last years, the use of the Segway® transporters for sightseeing tours in Vienna has increased distinctly, resulting in a growing number of Segway® related injuries and subsequent admissions of these patients to the Lorenz Böhler Trauma Centre in Vienna, Austria. METHODS: A retrospective analysis of clinical records in the electronic data system of the LBTC in Vienna, Austria, was performed to identify Segway® transporter related injuries between January 2010 and December 2012. RESULTS: Eighty-six patients represented the study cohort. The median age was 38 years (range 14-80 years) with a majority of male patients. Most common injuries were contusions (24, 6 %, n = 44) and fractures (23, 5 %, n = 42). The most frequent injury was a fracture of the radial head in 15, 1 % of all patients. 13 (15, 1 %) of 86 patients required admission and seven (8, 1 %) of these 13 patients had surgical treatment. CONCLUSION: This case series presents severe injuries related to the use of a Segway® transporter. As a consequence, it has to be ensured that public tour operators need to provide sufficient safety instructions and equipment for people who are unfamiliar with riding a Segway® .

Speckle-based x-ray phase-contrast and dark-field imaging with a laboratory source.

We report on the observation and application of near-field speckles with a laboratory x-ray source. The detection of speckles is possible thanks to the enhanced brilliance properties of the used liquid-metal-jet source, and opens the way to a range of new applications in laboratory-based coherent x-ray imaging. Here, we use the speckle pattern for multimodal imaging of demonstrator objects. Moreover, we introduce algorithms for phase and dark-field imaging using speckle tracking, and we show that they yield superior results with respect to existing methods.

X-ray phase contrast with injected gas for tumor microangiography.

We show that the microvasculature of mouse tumors can be visualized using propagation-based phase-contrast x-ray imaging with gas as the contrast agent. The large density difference over the gas-tissue interface provides high contrast, allowing the imaging of small-diameter blood vessels with relatively short exposure times and low dose using a compact liquid-metal-jet x-ray source. The method investigated is applied to tumors (E1A/Ras-transformed mouse embryonic fibroblasts) grown in mouse ears, demonstrating sub-15-µm-diameter imaging of their blood vessels. The exposure time for a 2D projection image is a few seconds and a full tomographic 3D map takes some minutes. The method relies on the strength of the vasculature to withstand the gas pressure. Given that tumor vessels are known to be more fragile than normal vessels, we investigate the tolerance of the vasculature of 12 tumors to gas injection and find that a majority withstand 200 mbar pressures, enough to fill 12-µm-diameter vessels with gas. A comparison of the elasticity of tumorous and non-tumorous vessels supports the assumption of tumor vessels being more fragile. Finally, we conclude that the method has the potential to be extended to the imaging of 15 µm vessels in thick tissue, including mouse imaging, making it of interest for, e.g., angiogenesis research.

[Radiological evaluation of k-wire osteosynthesis, in comparison to fixed-angle-plate osteosynthesis, in patients aged 80 years or more with distal radius fractures]. / Radiologische Ergebnisse von K-Draht Osteosynthesen im Vergleich zu winkelstabilen Plattenosteosynthesen distaler Radiusfrakturen bei Patienten über 80 Lebensjahre.

BACKGROUND: This study evaluates the short-term radiological outcome of K-wire osteosynthesis (KWO) in comparison to the fixed-angle-plate osteosynthesis (ORIF) on distal radius fractures in elderly patients (aged 80 years or more) with osteoporotic bones. PATIENTS AND METHODS: This study retrospectivly compares the postoperative X-rays of distal radius fractures (obtained between the years of 1998-2009) of patients aged 80 years and above treated with KWO (228 fractures, mean age 85 years), with the results of patients who were treated with fixed-angle plate ORIF (120 fractures, mean age 84 years). Within the KWO results, we also further compared the radiological results of a static and a dynamic (Kapandji) KWO technique. Only patients with a postoperative, anatomic reduction, and those who were radiologically followed up in a period of 2 months and above were included. The radiological criteria included the palmar and radial inclination as well as the radial shortening. RESULTS: With KWO performed in a static technique, 24% of the postoperative results showed no reduction loss. The use of the dynamic Kapandji technique KWO, improved the positive results to 63%. However, almost a third of the fractures (30%) treated with KWO, had shifted back to their preoperative positions, or worsened overall. The fixed angle plate (ORIF) was able to maintain 76% of all fractures in their postoperative positions. Merely 1.7% of the ORIF group sustained a complete reduction loss. The fixed-angle plate osteosynthesis shows a significant decrease of cases in which a complete repositioning loss is experienced. CONCLUSION: Although the importance of anatomic reconstruction of distal radius fractures is often debated in cases involving elderly patients, it is our considered opinion that, should an operative solution be chosen, one should consider the fixed-angle-plate osteosynthesis as the preferred operation method to prevent loss of reduction.

Comparison of two x-ray phase-contrast imaging methods with a microfocus source.

We present a comparison for high-resolution imaging with a laboratory source between grating-based (GBI) and propagation-based (PBI) x-ray phase-contrast imaging. The comparison is done through simulations and experiments using a liquid-metal-jet x-ray microfocus source. Radiation doses required for detection in projection images are simulated as a function of the diameter of a cylindrical sample. Using monochromatic radiation, simulations show a lower dose requirement for PBI for small object features and a lower dose for GBI for larger object features. Using polychromatic radiation, such as that from a laboratory microfocus source, experiments and simulations show a lower dose requirement for PBI for a large range of feature sizes. Tested on a biological sample, GBI shows higher noise levels than PBI, but its advantage of quantitative refractive index reconstruction for multi-material samples becomes apparent.

High average brightness water window source for short-exposure cryomicroscopy.

Laboratory water window cryomicroscopy has recently demonstrated similar image quality as synchrotron-based microscopy but still with much longer exposure times, prohibiting the spread to a wider scientific community. Here we demonstrate high-resolution laboratory water window imaging of cryofrozen cells with 10 s range exposure times. The major improvement is the operation of a λ=2.48 nm, 2 kHz liquid nitrogen jet laser plasma source with high spatial and temporal stability at high average brightness >1.5×10(12) ph/(s×sr×µm(2)×line), i.e., close to that of early synchrotrons. Thus, this source enables not only biological x-ray microscopy in the home laboratory but potentially other applications previously only accessible at synchrotron facilities.

Compact x-ray microscope for the water window based on a high brightness laser plasma source.

We present a laser plasma based x-ray microscope for the water window employing a high-average power laser system for plasma generation. At 90 W laser power a brightness of 7.4 x 10(11) photons/(s x sr x µm(2)) was measured for the nitrogen Lyα line emission at 2.478 nm. Using a multilayer condenser mirror with 0.3 % reflectivity 10(6) photons/(µm(2) x s) were obtained in the object plane. Microscopy performed at a laser power of 60 W resolves 40 nm lines with an exposure time of 60 s. The exposure time can be further reduced to 20 s by the use of new multilayer condenser optics and operating the laser at its full power of 130 W.

X-ray phase-contrast CO(2) angiography for sub-10 µm vessel imaging.

X-ray in-line phase contrast has recently been combined with CO(2) angiography for high-resolution small-animal vascular imaging at low radiation dose. In this paper we further investigate the potential and limitations of this method and demonstrate observation of vessels down to 8 µm in diameter, considerably smaller than the 60 µm previously reported. Our in-line phase-contrast imaging system is based on a liquid-metal-jet-anode x-ray source and utilizes free-space propagation to convert phase shifts, caused by refractive index variations, into intensity differences. Enhanced refractive index variations are obtained through injection of CO(2) gas into the vascular system to replace the blood. We show rat-kidney images with blood vessels down to 27 µm in diameter and mouse-ear images with vessels down to 8 µm. The minimum size of observable blood vessels is found to be limited by the penetration of gas into the vascular system and the signal-to-noise ratio, i.e. the allowed dose. The diameters of vessels being gas-filled depend on the gas pressure and follow a simple model based on surface tension. A theoretical signal-to-noise comparison shows that this method requires 1000 times less radiation dose than conventional iodine-based absorption contrast for observing sub-50 µm vessels.

X-ray phase contrast for CO2 microangiography.

We demonstrate a laboratory method for imaging small blood vessels using x-ray propagation-based phase-contrast imaging and carbon dioxide (CO(2)) gas as a contrast agent. The limited radiation dose in combination with CO(2) being clinically acceptable makes the method promising for small-diameter vascular visualization. We investigate the possibilities and limitations of the method for small-animal angiography and compare it with conventional absorption-based x-ray angiography. Photon noise in absorption-contrast imaging prevents visualization of blood vessels narrower than 50 µm at the highest radiation doses compatible with living animals, whereas our simulations and experiments indicate the possibility of visualizing 20 µm vessels at radiation doses as low as 100 mGy. Experimental computed tomography of excised rat kidney shows blood vessels of diameters down to 60 µm with improved image quality compared to absorption-based methods. With our present prototype x-ray source, the acquisition time for a tomographic dataset is approximately 1 h, which is long compared to the 1-20 min common for absorption-contrast micro-CT systems. Further development of the liquid-metal-jet microfocus x-ray sources used here and high-resolution x-ray detectors shows promise to reduce exposure times and make this high-resolution method practical for imaging of living animals.

Laboratory cryo soft X-ray microscopy.

Lens-based water-window X-ray microscopy allows two- and three-dimensional (2D and 3D) imaging of intact unstained cells in their near-native state with unprecedented contrast and resolution. Cryofixation is essential to avoid radiation damage to the sample. Present cryo X-ray microscopes rely on synchrotron radiation sources, thereby limiting the accessibility for a wider community of biologists. In the present paper we demonstrate water-window cryo X-ray microscopy with a laboratory-source-based arrangement. The microscope relies on a λ=2.48-nm liquid-jet high-brightness laser-plasma source, normal-incidence multilayer condenser optics, 30-nm zone-plate optics, and a cryo sample chamber. We demonstrate 2D imaging of test patterns, and intact unstained yeast, protozoan parasites and mammalian cells. Overview 3D information is obtained by stereo imaging while complete 3D microscopy is provided by full tomographic reconstruction. The laboratory microscope image quality approaches that of the synchrotron microscopes, but with longer exposure times. The experimental image quality is analyzed from a numerical wave-propagation model of the imaging system and a path to reach synchrotron-like exposure times in laboratory microscopy is outlined.

A 24 keV liquid-metal-jet x-ray source for biomedical applications.

We present a high-brightness 24-keV electron-impact microfocus x-ray source based on continuous operation of a heated liquid-indium/gallium-jet anode. The 30-70 W electron beam is magnetically focused onto the jet, producing a circular 7-13 µm full width half maximum x-ray spot. The measured spectral brightness at the 24.2 keV In K(α) line is 3 × 10(9) photons∕(s × mm(2) × mrad(2) × 0.1% BW) at 30 W electron-beam power. The high photon energy compared to existing liquid-metal-jet sources increases the penetration depth and allows imaging of thicker samples. The applicability of the source in the biomedical field is demonstrated by high-resolution imaging of a mammography phantom and a phase-contrast angiography phantom.

A 9 keV electron-impact liquid-gallium-jet x-ray source.

We demonstrate a high-brightness compact 9 keV electron-impact microfocus x-ray source based on a liquid-gallium-jet anode. A approximately 30 W, 50 kV electron gun is focused onto the approximately 20 ms, 30 mum diameter liquid-gallium-jet anode to produce an approximately 10 microm full width at half maximum x-ray spot. The peak spectral brightness is >2 x 10(10) photons(s mm(2) mrad(2)x0.1% BW). Calculation and experiments show potential for increasing this brightness by approximately three orders of magnitude, making the source suitable for laboratory-scale x-ray crystallography and hard x-ray microscopy.

Nerve regeneration using tubular scaffolds from biodegradable polyurethane.

INTRODUCTION: In severe nerve lesion, nerve defects and in brachial plexus reconstruction, autologous nerve grafting is the golden standard. Although, nerve grafting technique is the best available approach a major disadvantages exists: there is a limited source of autologous nerve grafts. This study presents data on the use of tubular scaffolds with uniaxial pore orientation from experimental biodegradable polyurethanes coated with fibrin sealant to regenerate a 8 mm resected segment of rat sciatic nerve. METHODS: Tubular scaffolds: prepared by extrusion of the polymer solution in DMF into water coagulation bath. The polymer used for the preparation of tubular scaffolds was a biodegradable polyurethane based on hexamethylene diisocyanate, poly(epsilon-caprolactone) and dianhydro-D-sorbitol. EXPERIMENTAL MODEL: Eighteen Sprague Dawley rats underwent mid-thigh sciatic nerve transection and were randomly assigned to two experimental groups with immediate repair: (1) tubular scaffold, (2) 180 degrees rotated sciatic nerve segment (control). Serial functional measurements (toe spread test, placing tests) were performed weekly from 3rd to 12th week after nerve repair. On week 12, electrophysiological assessment was performed. Sciatic nerve and scaffold/nerve grafts were harvested for histomorphometric analysis. Collagenic connective tissue, Schwann cells and axons were evaluated in the proximal nerve stump, the scaffold/nerve graft and the distal nerve stump. The implants have uniaxially-oriented pore structure with a pore size in the range of 2 micorm (the pore wall) and 75 x 700 microm (elongated pores in the implant lumen). The skin of the tubular implants was nonporous. Animals which underwent repair with tubular scaffolds of biodegradable polyurethanes coated with diluted fibrin sealant had no significant functional differences compared with the nerve graft group. Control group resulted in a trend-wise better electrophysiological recovery but did not show statistically significant differences. There was a higher level of collagenic connective tissue within the scaffold and within the distal nerve stump. Schwann cells migrated into the polyurethane scaffold. There was no statistical difference to the nerve graft group although Schwann cell counts were lower especially within the middle of the polyurethane scaffold. Axon counts showed a trend-wise decrease within the scaffold. CONCLUSION: These results suggest that biodegradable polyurethane tubular scaffolds coated with diluted fibrin sealant support peripheral nerve regeneration in a standard gap model in the rat up to 3 months. Three months after surgery no sign of degradation could be seen.

[Gunshot injuries--incidence and treatment]. / Schussverletzungen--ihre Inzidenz und Versorgung.

BACKGROUND: Gunshot injuries are very rare in the European Union. To show the requirements for the trauma surgeon to deal with gunshot injuries this analysis was performed. METHODS: Gunshot injuries seen at Lorenz Boehler Trauma Center from 1997 to 2004 were reviewed. The case histories of 67 patients were analysed for the cause of the gunshot injury, type of weapon, surgical intervention, days of treatment and outcome. RESULTS: The most commonly cause of gunshot injuries were criminal offenses (n = 35). Handguns were used most often (n = 32). The locations of injuries were evenly distributed over head (n = 18 ), trunk (n = 16), upper (n = 15) and lower limb (n = 18). Surgical treatment was performed in 57 patients. The median stay on ICU was 10 days. 6 patients died. CONCLUSION: Gunshot injuries are frequently not confined to the extremities. The challenge for the trauma surgeon is to deal with injuries of the head, chest and abdomen.

High-resolution compact X-ray microscopy.

We demonstrate compact full-field soft X-ray transmission microscopy with sub 60-nm resolution operating at lambda= 2.48 nm. The microscope is based on a 100-Hz regenerative liquid-nitrogen-jet laser-plasma source in combination with a condenser zone plate and a micro-zone plate objective for high-resolution imaging onto a 2048 x 2048 pixel CCD detector. The sample holder is mounted in a helium atmosphere and allows imaging of both dry and wet specimens. The microscope design enables fast sample switching and the sample can be pre-aligned using a visible-light microscope. High-quality images can be acquired with exposure times of less than 5 min. We demonstrate the performance of the microscope using both dry and wet samples.

A vacuum-compatible wet-specimen chamber for compact X-ray microscopy.

Soft X-ray microscopy is a powerful tool for investigations of, for example, polymers or soils in their natural liquid environment. This requires a wet-specimen chamber. Compact X-ray microscopy allows the horizontal mounting of such samples, thereby reducing the influence of gravitational forces. We have developed a wet-specimen chamber for such compact X-ray microscope. The chamber is vacuum compatible, which reduces the exposure time. The vacuum sealing is achieved by a combination of mechanical sealing and sealing by bio-compatible glue. With the wet-specimen chamber the specimens can be kept in an aqueous environment in a vacuum of 10(-4) mbar for several hours. Imaging of lipid droplets in water demonstrates the function of the wet-specimen chamber.

Size-selective colloidal-gold localization in transmission X-ray microscopy.

Colloidal gold is a useful marker for functional-imaging experiments in transmission X-ray microscopy. Due to the low contrast of gold particles with small diameters it is necessary to develop a powerful algorithm to localize the single gold particles. The presented image-analysis algorithm for identifying colloidal gold particles is based on the combination of a threshold with respect to the local absorption and shape discrimination, realized by fitting a Gaussian profile to the identified regions of interest. The shape discrimination provides the possibility of size-selective identification and localization of single colloidal gold particles down to a diameter of 50 nm. The image-analysis algorithm, therefore, has potential for localization studies of several proteins simultaneously and for localization of fiducial markers in X-ray tomography.

Proliferation and viability of adherent cells manipulated by standing-wave ultrasound in a microfluidic chip.

Ultrasonic-standing-wave (USW) technology has potential to become a standard method for gentle and contactless cell handling in microfluidic chips. We investigate the viability of adherent cells exposed to USWs by studying the proliferation rate of recultured cells following ultrasonic trapping and aggregation of low cell numbers in a microfluidic chip. The cells form 2-D aggregates inside the chip and the aggregates are held against a continuous flow of cell culture medium perpendicular to the propagation direction of the standing wave. No deviations in the doubling time from expected values (24 to 48 h) were observed for COS-7 cells held in the trap at acoustic pressure amplitudes up to 0.85 MPa and for times ranging between 30 and 75 min. Thus, the results demonstrate the potential of ultrasonic standing waves as a tool for gentle manipulation of low cell numbers in microfluidic systems.

Ultrasonic enhancement of bead-based bioaffinity assays.

Ultrasonic radiation forces can be used for non-intrusive manipulation and concentration of suspended micrometer-sized particles. For bioanalytical purposes, standing-wave ultrasound has long been used for rapid immuno-agglutination of functionalized latex beads. More recently, detection methods based on laser-scanning fluorometry and single-step homogeneous bead-based assays show promise for fast, easy and sensitive biochemical analysis. If such methods are combined with ultrasonic enhancement, detection limits in the femtomolar region are feasible. In this paper, we review the development of standing-wave ultrasonic manipulation for bioanalysis, with special emphasis on miniaturization and ultrasensitive bead-based immunoassays.