MRI overestimates articular cartilage thickness and volume compared to synchrotron radiation phase-contrast imaging.

Accurate evaluation of morphological changes in articular cartilage are necessary for early detection of osteoarthritis (OA). 3T magnetic resonance imaging (MRI) has highly sensitive contrast resolution and is widely used clinically to detect OA. However, synchrotron radiation phase-contrast imaging computed tomography (SR-PCI) can also provide contrast to tissue interfaces that do not have sufficient absorption differences, with the added benefit of very high spatial resolution. Here, MRI was compared with SR-PCI for quantitative evaluation of human articular cartilage. Medial tibial condyles were harvested from non-OA donors and from OA patients receiving knee replacement surgery. Both imaging methods revealed that average cartilage thickness and cartilage volume were significantly reduced in the OA group, compared to the non-OA group. When comparing modalities, the superior resolution of SR-PCI enabled more precise mapping of the cartilage surface relative to MRI. As a result, MRI showed significantly higher average cartilage thickness and cartilage volume, compared to SR-PCI. These data highlight the potential for high-resolution imaging of articular cartilage using SR-PCI as a solution for early OA diagnosis. Recognizing current limitations of using a synchrotron for clinical imaging, we discuss its nascent utility for preclinical models, particularly longitudinal studies of live animal models of OA.

Microbotanical residues for the study of early hominin tools.

More than 2 million years ago in East Africa, the earliest hominin stone tools evolved amidst changes in resource base, with pounding technology playing a key role in this adaptive process. Olduvai Gorge (now Oldupai) is a famed locality that remains paramount for the study of human evolution, also yielding some of the oldest battering tools in the world. However, direct evidence of the resources processed with these technologies is lacking entirely. One way to obtain this evidence is through the analysis of surviving residues. Yet, linking residues with past processing activities is not simple. In the case of plant exploitation, this link can only be established by assessing site-based reference collections inclusive of both anthropogenic and natural residues as a necessary first step and comparative starting point. In this paper, we assess microbotanical remains from rock clasts sourced at the same quarry utilized by Oldowan hominins at Oldupai Gorge. We mapped this signal and analysed it quantitatively to classify its spatial distribution objectively, extracting proxies for taxonomic identification and further comparison with freestanding soils. In addition, we used blanks to manufacture pounding tools for blind, controlled replication of plant processing. We discovered that stone blanks are in fact environmental reservoirs in which plant remains are trapped by lithobionts, preserved as hardened accretions. Tool use, on the other hand, creates residue clusters; however, their spatial distribution can be discriminated from purely natural assemblages by the georeferencing of residues and statistical analysis of resulting patterns. To conclude, we provide a protocol for best practice and a workflow that has the advantage of overcoming environmental noise, reducing the risk of false positive, delivering a firm understanding of residues as polygenic mixtures, a reliable use of controls, and most importantly, a stronger link between microbotanical remains and stone tool use.

Timing of Mouse Molar Formation Is Independent of Jaw Length Including Retromolar Space.

For humans and other mammals to eat effectively, teeth must develop properly inside the jaw. Deciphering craniodental integration is central to explaining the timely formation of permanent molars, including third molars which are often impacted in humans, and to clarifying how teeth and jaws fit, function and evolve together. A factor long-posited to influence molar onset time is the jaw space available for each molar organ to form within. Here, we tested whether each successive molar initiates only after a minimum threshold of space is created via jaw growth. We used synchrotron-based micro-CT scanning to assess developing molars in situ within jaws of C57BL/6J mice aged E10 to P32, encompassing molar onset to emergence. We compared total jaw, retromolar and molar lengths, and molar onset times, between upper and lower jaws. Initiation time and developmental duration were comparable between molar upper and lower counterparts despite shorter, slower-growing retromolar space in the upper jaw, and despite size differences between upper and lower molars. Timing of molar formation appears unmoved by jaw length including space. Conditions within the dental lamina likely influence molar onset much more than surrounding jaw tissues. We theorize that molar initiation is contingent on sufficient surface area for the physical reorganization of dental epithelium and its invagination of underlying mesenchyme.

The effects of sodium reduction on the gas phase of bread doughs using synchrotron X-ray microtomography.

Globally, the bakery industry has a target of reducing sodium content in bread products. However, removing salt results in changes in the quality of bread through effects on dough's gas phase during the breadmaking process. Using synchrotron X-ray microtomography, the objective of this study was to investigate how sodium reduction induced changes in the gas phase parameters (i.e., gas volume fraction, bubble size distribution (BSD) and its time evolution) of non-yeasted doughs made from a wide range of formulations (i.e., wheat cultivar and water content) prepared with different mixing times. As salt content was reduced, a lower gas volume was retained in the dough by the end of mixing. Less gas bubbles were also retained if doughs were prepared from a stronger wheat cultivar, higher water content, and/or mixed for a shorter time. Rates of change in the median (R0) and the width (ε) of the fitted lognormal radius dependence of bubble volume fraction [BVF(R)] indicated that reduced sodium content permitted disproportionation to proceed more rapidly. Higher water content or longer mixing time also resulted in faster disproportionation, indicating that water content and mixing time can be manipulated as a means of increasing bubble stability against disproportionation during low-sodium breadmaking. An examination of relative changes in dough's gas phase parameters arising from sodium reduction demonstrated that wheat cultivar, water content and mixing time all affected dough's tolerance to sodium reduction. Therefore, attainment of good bread crumb cell structure in low-sodium bread formulas is a function of salt's effects on dough rheology in addition to its effect on yeast activity, so that dough formulation and mixing conditions also need to be considered.

Application of Two-Photon-Absorption Pulsed Laser for Single-Event-Effects Sensitivity Mapping Technology.

Single-event effects (SEEs) in integrated circuits and devices can be studied by utilizing ultra-fast pulsed laser system through Two Photon Absorption process. This paper presents technical ways to characterize key factors for laser based SEEs mapping testing system: output power from laser source, spot size focused by objective lens, opening window of Pockels cell, and calibration of injected laser energy. The laser based SEEs mapping testing system can work in a stable and controllable status by applying these methods. Furthermore, a sensitivity map of a Static Random Access Memory (SRAM) cell with a 65 nm technique node was created through the established laser system. The sensitivity map of the SRAM cell was compared to a map generated by a commercial simulation tool (TFIT), and the two matched well. In addition, experiments in this paper also provided energy distribution profile along Z axis that is the direction of the pulsed laser injection and threshold energy for different SRAM structures.

Protocol development for synchrotron contrast-enhanced CT of human hip cartilage.

Understanding hip osteoarthritis requires new investigational tools for quantitative studies of biophysical and biomechanical properties as well as for determination of structure. Three new protocols to study pathological changes in cartilage and to measure cartilage thickness in intact human hips are described using synchrotron contrast enhanced computed tomography (sCECT) with the iodinated contrast agent CA4+. Ten human cadaver hips were prepared and injected with CA4+ using three different methods, all of which included rotation and distraction of the joint. CA4+ diffusion into cartilage was monitored using sCECT. The thickness of acetabular and femoral cartilage was also measured. Diffusion times ranged from 2 h to 75 h, depending on the injection protocol and the cartilage region. Direct single injection of the contrast through the labrum resulted in the fastest diffusion times. The iodine attenuation coefficient, which reflects the contrast agent distribution in the cartilage, ranged from 0.0142/cm to 0.1457/cm. Three injections at the head/neck conjunction area yielded the highest iodine attenuation coefficients in cartilage. The femoral cartilage in the Superior-Medial compartment was significantly thicker than in the other 3 femoral compartments, and femoral cartilage in the Superior-Anterior compartment was significantly thinner than the other 3 femoral compartments. The acetabular cartilage in the Superior compartment was significantly thicker than that in the Superior-Posterior compartment. sCECT with CA4+ allows assessment of hip cartilage thickness with 0.1 mm isotropic voxel size, sufficient for evaluating cartilage pathology and biomechanics.

Development and assessment of a multi-beam continuous-phantom-motion x-ray scatter projection imaging system.

X-ray image formation using scattered radiation can yield a superior contrast-to-noise ratio compared to conventional transmission x-ray imaging. A barrier to practical implementation of scatter imaging systems has been slow image acquisition. We have developed a projection imaging system which uses five monoenergetic pencil beams in combination with continuous phantom motion to achieve acquisition times that are practical for medical and security applications. The system was configured at the Canadian Light Source synchrotron and consists of a primary collimator, motorized stages for phantom translation, a flat-panel x-ray detector for measuring scattered x rays, and photodiodes for simultaneously measuring transmitted x rays. Image generation requires several corrections to raw data artifacts arising from the nature of the detector, x-ray source, and acquisition procedure. We developed a novel correction for pixel location inaccuracy arising from continuous phantom motion. A five-beam system had nearly five times faster acquisition than a single-beam system. Continuous motion acquisition was approximately 30 times faster than step-and-shoot acquisition. The total acquisition time for a 9 cm × 5 cm phantom with 8425 pixels was just over 2 min. Image quality was also assessed, in part to determine its relation to acquisition speed. The width of sharp material boundaries was found to be at a minimum equal to the pencil beam width (1.75 mm) and to have an additional width equal to the product of the phantom translation speed and the acquisition time per pixel (up to 1.0 mm in our experiments). Contrast-detail performance was independent of acquisition speed, depending only on phantom entrance x-ray fluence. Pixel signal-to-noise ratio measurements indicate that detector readout noise is important for the scatter data, even for phantom air kerma as high as 30 mGy. Images could be improved with a detector having lower readout noise and higher sensitivity. Its spatial resolution could be moderate. We confirmed that for the same range of λ-1 sin(θ/2), where λ is the x-ray wavelength and θ is the scattering angle, scatter images acquired using different beam energies (33-70 keV) had nearly identical contrast.

Nebulized hypertonic saline triggers nervous system-mediated active liquid secretion in cystic fibrosis swine trachea.

Inhaled hypertonic saline (HTS) treatment is used to improve lung health in patients with cystic fibrosis (CF). The current consensus is that the treatment generates an osmotic gradient that draws water into the airways and increases airway surface liquid (ASL) volume. However, there is evidence that HTS may also stimulate active secretion of ASL by airway epithelia through the activation of sensory neurons. We tested the contribution of the nervous system and airway epithelia on HTS-stimulated ASL height increase in CF and wild-type swine airway. We used synchrotron-based imaging to investigate whether airway neurons and epithelia are involved in HTS treatment-triggered ASL secretion in CFTR-/- and wild-type swine. We showed that blocking parasympathetic and sensory neurons in airway resulted in ~50% reduction of the effect of HTS treatment on ASL volume in vivo. Incubating tracheal preparations with inhibitors of epithelial ion transport across airway decreased secretory responses to HTS treatment. CFTR-/- swine ex-vivo tracheal preparations showed substantially decreased secretory response to HTS treatment after blockage of neuronal activity. Our results indicated that HTS-triggered ASL secretion is partially mediated by the stimulation of airway neurons and the subsequent activation of active epithelia secretion; osmosis accounts for only ~50% of the effect.

High-power-load DCLM monochromator for a computed tomography program at BMIT at energies of 25-150 keV.

The research program at the biomedical imaging facility requires a high-flux hard-X-ray monochromator that can also provide a wide beam. A wide energy range is needed for standard radiography, phase-contrast imaging, K-edge subtraction imaging and monochromatic beam therapy modalities. The double-crystal Laue monochromator, developed for the BioMedical Imaging and Therapy facility, is optimized for the imaging of medium- and large-scale samples at high energies with the resolution reaching 4 µm. A pair of 2 mm-thick Si(111) bent Laue-type crystals were used in fixed-exit beam mode with a 16 mm vertical beam offset and the first crystal water-cooled. The monochromator operates at energies from 25 to 150 keV, and the measured size of the beam is 189 mm (H) × 8.6 mm (V) at 55 m from the source. This paper presents our approach in developing a complete focusing model of the monochromator. The model uses mechanical properties of crystals and benders to obtain a finite-element analysis of the complete assembly. The modeling results are compared and calibrated with experimental measurements. Using the developed analysis, a rough estimate of the bending radius and virtual focus (image) position of the first crystal can be made, which is also the real source for the second crystal. On the other hand, by measuring the beam height in several points in the SOE-1 hutch, the virtual focus of the second crystal can be estimated. The focusing model was then calibrated with measured mechanical properties, the values for the force and torque applied to the crystals were corrected, and the actual operating parameters of the monochromator for fine-tuning were provided.

Countercurrent heat exchange and thermoregulation during blood-feeding in kissing bugs.

Blood-sucking insects experience thermal stress at each feeding event on endothermic vertebrates. We used thermography to examine how kissing-bugs Rhodnius prolixus actively protect themselves from overheating. During feeding, these bugs sequester and dissipate the excess heat in their heads while maintaining an abdominal temperature close to ambient. We employed a functional-morphological approach, combining histology, µCT and X-ray-synchrotron imaging to shed light on the way these insects manage the flow of heat across their bodies. The close alignment of the circulatory and ingestion systems, as well as other morphological characteristics, support the existence of a countercurrent heat exchanger in the head of R. prolixus, which decreases the temperature of the ingested blood before it reaches the abdomen. This kind of system has never been described before in the head of an insect. For the first time, we show that countercurrent heat exchange is associated to thermoregulation during blood-feeding.

Superhydrophobicity from the Inside.

The nature of trapped air on submersed ultra-water-repellent interfaces has been investigated. These gaseous layers (plastrons) can last from hours to, in some examples such as the Salvinia molesta fern, months. The interface of submerged superhydrophobic surfaces with carefully controlled micropatterned surface roughness has been probed using synchrotron-based high-resolution X-ray phase tomography. This technique looks in situ, through the aqueous/gas interface in three dimensions. Long-term plastron stability appears to correlate with the appearance of scattered microdroplets <20 µm in diameter that are sandwiched within the 30 µm thick gaseous interfacial layer. These microdroplets are centered on defects or damaged sections within the substrate surface approximately 20-50 µm apart. Such irregularities represent heterogeneous micro/nano-hierarchical structures with varying surface structures and chemistry. The stability of microdroplets is governed by a combination of electrostatic repulsion, contact angle limitations, and a saturated vapor pressure, the latter of which reduces the rate of diffusion of gas out of the air layer, thus increasing underwater longevity. Homogenous surfaces exhibiting purely nano- or micro-regularity do not support such microdroplets, and, as a consequence, plastrons can disappear in <20 h compared with >160 h for surfaces with scattered microdroplets. Such behavior may be a requirement for long-term nonwetting in any system.

Cystic fibrosis swine fail to secrete airway surface liquid in response to inhalation of pathogens.

Cystic fibrosis is caused by mutations in the gene encoding the cystic fibrosis transmembrane conductance regulator (CFTR) channel, which can result in chronic lung disease. The sequence of events leading to lung disease is not fully understood but recent data show that the critical pathogenic event is the loss of the ability to clear bacteria due to abnormal airway surface liquid secretion (ASL). However, whether the inhalation of bacteria triggers ASL secretion and whether this is abnormal in cystic fibrosis has never been tested. Here we show, using a novel synchrotron-based in vivo imaging technique, that wild-type pigs display both a basal and a Toll-like receptor-mediated ASL secretory response to the inhalation of cystic fibrosis relevant bacteria. Both mechanisms fail in CFTR-/- swine, suggesting that cystic fibrosis airways do not respond to inhaled pathogens, thus favoring infection and inflammation that may eventually lead to tissue remodeling and respiratory disease.Cystic fibrosis is caused by mutations in the CFTR chloride channel, leading to reduced airway surface liquid secretion. Here the authors show that exposure to bacteria triggers secretion in wild-type but not in pig models of cystic fibrosis, suggesting an impaired response to pathogens contributes to infection.

Three-dimensional labeling of newly formed bone using synchrotron radiation barium K-edge subtraction imaging.

Bone is a dynamic tissue which exhibits complex patterns of growth as well as continuous internal turnover (i.e. remodeling). Tracking such changes can be challenging and thus a high resolution imaging-based tracer would provide a powerful new perspective on bone tissue dynamics. This is, particularly so if such a tracer can be detected in 3D. Previously, strontium has been demonstrated to be an effective tracer which can be detected by synchrotron-based dual energy K-edge subtraction (KES) imaging in either 2D or 3D. The use of strontium is, however, limited to very small sample thicknesses due to its low K-edge energy (16.105 keV) and thus is not suitable for in vivo application. Here we establish proof-of-principle for the use of barium as an alternative tracer with a higher K-edge energy (37.441 keV), albeit for ex vivo imaging at the moment, which enables application in larger specimens and has the potential to be developed for in vivo imaging of preclinical animal models. New bone formation within growing rats in 2D and 3D was demonstrated at the Biomedical Imaging and Therapy bending magnet (BMIT-BM) beamline of the Canadian Light Source synchrotron. Comparative x-ray fluorescence imaging confirmed those patterns of uptake detected by KES. This initial work provides a platform for the further development of this tracer and its exploration of applications for in vivo development.

Data of low-dose phase-based X-ray imaging for in situ soft tissue engineering assessments.

This article presents the data of using three phase-based X-ray imaging techniques to characterize biomaterial scaffold and soft tissues in situ, as reported in our study "Low-dose phase-based X-ray imaging techniques for in situ soft tissue engineering assessments" [1]. The examined parameters include the radiation dose, scan time, and image quality, which are all critical to longitudinal in situ live animal assessments. The data presented were obtained from three dimensional imaging of scaffolds in situ cartilage by means of synchrotron-based computed tomography-diffraction enhanced imaging (CT-DEI), analyzer based imaging (CT-ABI), and in-line phase contrast imaging (CT-PCI) at standard and low dose imaging modalities.

Low-dose phase-based X-ray imaging techniques for in situ soft tissue engineering assessments.

In tissue engineering, non-invasive imaging of biomaterial scaffolds and tissues in living systems is essential to longitudinal animal studies for assessments without interrupting the repair process. Conventional X-ray imaging is inadequate for use in soft tissue engineering due to the limited absorption difference between the soft tissue and biomaterial scaffolds. X-ray phase-based imaging techniques that derive contrast from refraction or phase effects rather than absorption can provide the necessary contrast to see low-density biomaterial scaffolds and tissues in large living systems. This paper explores and compares three synchrotron phase-based X-ray imaging techniques-computed tomography (CT)-diffraction enhanced imaging (DEI), -analyzer based imaging (ABI), and -phase contrast imaging (PCI)-for visualization and characterization of low-density biomaterial scaffolds and tissues in situ for non-invasive soft tissue engineering assessments. Intact pig joints implanted with polycaprolactone scaffolds were used as the model to assess and compare the imaging techniques in terms of different qualitative and quantitative criteria. For long-term in vivo live animal imaging, different strategies for reducing the imaging radiation dose and scan time-reduced number of CT projections, region of interest, and low resolution imaging-were examined with the presented phase-based imaging techniques. The results demonstrated promising capabilities of the phase-based techniques for visualization of biomaterial scaffolds and soft tissues in situ. The low-dose imaging strategies were illustrated effective for reducing the radiation dose to levels appropriate for live animal imaging. The comparison among the imaging techniques suggested that CT-DEI has the highest efficiency in retaining image contrast at considerably low radiation doses.

Application of analyzer based X-ray imaging technique for detection of ultrasound induced cavitation bubbles from a physical therapy unit.

BACKGROUND: The observation of ultrasound generated cavitation bubbles deep in tissue is very difficult. The development of an imaging method capable of investigating cavitation bubbles in tissue would improve the efficiency and application of ultrasound in the clinic. Among the previous imaging modalities capable of detecting cavitation bubbles in vivo, the acoustic detection technique has the positive aspect of in vivo application. However the size of the initial cavitation bubble and the amplitude of the ultrasound that produced the cavitation bubbles, affect the timing and amplitude of the cavitation bubbles' emissions. METHODS: The spatial distribution of cavitation bubbles, driven by 0.8835 MHz therapeutic ultrasound system at output power of 14 Watt, was studied in water using a synchrotron X-ray imaging technique, Analyzer Based Imaging (ABI). The cavitation bubble distribution was investigated by repeated application of the ultrasound and imaging the water tank. The spatial frequency of the cavitation bubble pattern was evaluated by Fourier analysis. Acoustic cavitation was imaged at four different locations through the acoustic beam in water at a fixed power level. The pattern of cavitation bubbles in water was detected by synchrotron X-ray ABI. RESULTS: The spatial distribution of cavitation bubbles driven by the therapeutic ultrasound system was observed using ABI X-ray imaging technique. It was observed that the cavitation bubbles appeared in a periodic pattern. The calculated distance between intervals revealed that the distance of frequent cavitation lines (intervals) is one-half of the acoustic wave length consistent with standing waves. CONCLUSION: This set of experiments demonstrates the utility of synchrotron ABI for visualizing cavitation bubbles formed in water by clinical ultrasound systems working at high frequency and output powers as low as a therapeutic system.

Design of a mouse restraint for synchrotron-based computed tomography imaging.

High-resolution computed tomography (CT) imaging of a live animal within a lead-lined synchrotron light hutch presents several unique challenges. In order to confirm that the animal is under a stable plane of anaesthesia, several physiological parameters (e.g. heart rate, arterial oxygen saturation, core body temperature and respiratory rate) must be remotely monitored from outside the imaging hutch. In addition, to properly scan the thoracic region using CT, the animal needs to be held in a vertical position perpendicular to the fixed angle of the X-ray beam and free to rotate 180°-360°. A new X-ray transparent mouse restraint designed and fabricated using computer-aided design software and three-dimensional rapid prototype printing has been successfully tested at the Biomedical Imaging and Therapy bending-magnet (BMIT-BM) beamline at the Canadian Light Source.

Diffraction-Enhanced Computed Tomographic Imaging of Growing Piglet Joints by Using a Synchrotron Light Source.

The objective of this project was to develop and test a new technology for imaging growing joints by means of diffraction-enhanced imaging (DEI) combined with CT and using a synchrotron radiation source. DEI-CT images of an explanted 4-wk-old piglet stifle joint were acquired by using a 40-keV beam. The series of scanned slices was later 'stitched' together, forming a 3D dataset. High-resolution DEI-CT images demonstrated fine detail within all joint structures and tissues. Striking detail of vasculature traversing between bone and cartilage, a characteristic of growing but not mature joints, was demonstrated. This report documents for the first time that DEI combined with CT and a synchrotron radiation source can generate more detailed images of intact, growing joints than can currently available conventional imaging modalities.

A phase-space beam position monitor for synchrotron radiation.

The stability of the photon beam position on synchrotron beamlines is critical for most if not all synchrotron radiation experiments. The position of the beam at the experiment or optical element location is set by the position and angle of the electron beam source as it traverses the magnetic field of the bend-magnet or insertion device. Thus an ideal photon beam monitor would be able to simultaneously measure the photon beam's position and angle, and thus infer the electron beam's position in phase space. X-ray diffraction is commonly used to prepare monochromatic beams on X-ray beamlines usually in the form of a double-crystal monochromator. Diffraction couples the photon wavelength or energy to the incident angle on the lattice planes within the crystal. The beam from such a monochromator will contain a spread of energies due to the vertical divergence of the photon beam from the source. This range of energies can easily cover the absorption edge of a filter element such as iodine at 33.17 keV. A vertical profile measurement of the photon beam footprint with and without the filter can be used to determine the vertical centroid position and angle of the photon beam. In the measurements described here an imaging detector is used to measure these vertical profiles with an iodine filter that horizontally covers part of the monochromatic beam. The goal was to investigate the use of a combined monochromator, filter and detector as a phase-space beam position monitor. The system was tested for sensitivity to position and angle under a number of synchrotron operating conditions, such as normal operations and special operating modes where the photon beam is intentionally altered in position and angle at the source point. The results are comparable with other methods of beam position measurement and indicate that such a system is feasible in situations where part of the synchrotron beam can be used for the phase-space measurement.

Factors influencing real time internal structural visualization and dynamic process monitoring in plants using synchrotron-based phase contrast X-ray imaging.

Minimally invasive investigation of plant parts (root, stem, leaves, and flower) has good potential to elucidate the dynamics of plant growth, morphology, physiology, and root-rhizosphere interactions. Laboratory based absorption X-ray imaging and computed tomography (CT) systems are extensively used for in situ feasibility studies of plants grown in natural and artificial soil. These techniques have challenges such as low contrast between soil pore space and roots, long X-ray imaging time, and low spatial resolution. In this study, the use of synchrotron (SR) based phase contrast X-ray imaging (PCI) has been demonstrated as a minimally invasive technique for imaging plants. Above ground plant parts and roots of 10 day old canola and wheat seedlings grown in sandy clay loam soil were successfully scanned and reconstructed. Results confirmed that SR-PCI can deliver good quality images to study dynamic and real time processes such as cavitation and water-refilling in plants. The advantages of SR-PCI, effect of X-ray energy, and effective pixel size to study plant samples have been demonstrated. The use of contrast agents to monitor physiological processes in plants was also investigated and discussed.