Imaging anatomical features of the human brain at cellular resolution currently relies on series of physical sections with related slicing artefacts. So far, microtomography has been employed to image an entire human brain at a voxel size of 20µm and selected regions using 6µm. This study aims to demonstrate the feasibility of imaging the entire human brain with cellular resolution without the need for physical sectioning using hard x-ray computed tomography. 1.2mm high sections of two human brains, one embedded in ethanol, the other in paraffin, were imaged using microtomography at the P07 beamline at DESY, Hamburg, Germany with a monochromatic beam at 67keV. The extended field of view necessary to cover the ca. 10 cm wide specimens at 2.54µm voxel size was realized by projection tiling with eight to ten rings. The resulting reconstructed slices measured 39,000×39,000 voxels. This synchrotron radiation-based study shows the feasibility of employing x-ray tomography to image the entire human brain with isotropic voxels of 2.54µm resolution. Next, we need to tackle the vertical stitching of several 10,000 slices of 6GB each, posing the challenge of processing the big data of an entire PB-sized human brain and making it accessible to the research community.
Transport of immune cells, nutrients and waste products via the cerebrospinal fluid (CSF) has been implicated in the development of neurological disorders. Using time-resolved in vivo microtomography, we investigated pulsatile motion of CSF spaces in the mouse brain as a potential driver of CSF flow. Here we present a method for detecting motion captured in murine brain images acquired in vivo at the European Synchrotron Radiation Facility. Anesthetized mice were placed in a heated holder that was designed to minimize head motion and maintain physiological body temperature. Contrast agent was infused into the ventricle to improve visibility of the CSF spaces. Projections were retrospectively sorted based on the ECG recording. Cardiac phase images were reconstructed in 10ms intervals from the ECG peak and automatically analyzed for decorrelation. Motion was automatically quantified by non-rigid registration. Regions with high intensity structures, large motion magnitudes, large improvements in image similarity due to registration, or at the contrast-enhanced ventricles were visually inspected for structures with motion artifacts prior to registration. We detected mainly motion in the nasopharynx, skin, ear channels, and bones in the range of 2.3 to 14.8µm magnitude. Small motion artifacts were detectable only for high-contrast structures. No misalignments were visible for the contrast-enhanced ventricles at a voxel resolution of 6.30 to 6.45µm. In the future, dedicated active gating to ensure regular sampling and local scans with improved spatial resolution will be used to investigate the limits to the detection of in vivo ventricular motion in mice.
The acquisition of large tomography volumes, exceeding the typical detector field-of-view, requires advanced acquisition techniques. Current approaches are the tiling of local reconstructed volumes or the tiling in projection space, also known as mosaic tomography. Reconstruction tiling has the advantage that standard reconstruction software can be used and acquisition can be interrupted and resumed relatively easily. The disadvantage is that there is the need for volume registration and transformation. Projection tiling is faster and more dose efficient, however a custom reconstruction pipeline is required, registration in projection space is challenging due to lower contrast, and there is a high sensitivity to mechanical instabilities. In this work we propose a third, hybrid approach, to profit from the advantages of projection tiling, but limit the risks. The volume to be imaged is covered by overlapping cylinders, each corresponding to the reconstructed volume of one mosaic tomogram. The number of rings per cylinder and the total number of cylinders can be tuned to the specimen at hand. We demonstrate this approach for a 2cm-wide section of a human brain stem, imaged at the Anatomix beamline of Synchrotron Soleil, France with 0.65µm voxel size, resulting in reconstructed slices 29,650 voxels wide. For mosaic reconstruction we used our team’s existing pipeline. For stitching of volumes, image registration was performed in the overlap regions. As pairwise displacements between cylinders are not independent, we modified the registration approach to force a consistent solution. The results of the hybrid acquisition in seven tiles with four rings were compared to a pure projection tiling approach with eight rings and to local regions representing reconstruction tiling. In conclusion, we propose an extended field of view acquisition scheme building on the speed and dose efficiency of mosaic acquisition, but relaxing the requirements for mechanical and beam stability.
Caries affects billions of individuals worldwide, thus pointing out the importance of advancements in restorative dentistry. Dental resin composites yield restorations with satisfying mechanical properties, therefore the focus of development has shifted to accelerated treatments and esthetic aspects. Challenges in matching tooth color arise due to limited options, application changes, and color variations over time. Single-shade composites with the 'chameleon effect' adapt their color to the surrounding enamel by closely matching the tooth's optical spectrum, enhancing color blending. Structural color, based on light interference, contributes to this effect. The study investigates the submicron filler particles' impact on optical properties and the chameleon effect. Four single-shade dental resin composite materials were investigated. Needle-like samples about 100 μm in diameter were prepared and imaged in a scanning electron microscope. Light transmission through the materials for wavelengths between 200 and 900 nm was measured using a spectrophotometer. Three-dimensional nanotomography data were obtained through transmission X-ray microscopy at the ANATOMIX beamline, Synchrotron SOLEIL, France in both absorption and Zernike phase contrast mode with 23 nm voxel size. The real space information was complemented with small-angle X-ray scattering. These experiments revealed substantial differences in the microscopic structure of the materials. In the case of Omnichroma, the filler consists of almost identical spheres with a diameter of 260 nm while Filtek Universal exhibits polydisperse, irregularly shaped fillers. Additionally, Venus Pearl One’s fillers have a polyhedral shape and a wide size distribution. Finally, the setups used did not reveal any clearly identified microstructure of the Chroma Fill composite. Although all investigated materials are known to exhibit the chameleon effect, their differences in micro- and nanostructure call into question previous hypotheses on the chameleon effect’s origin from structural color. While we have now a reasonable understanding of filler morphology, size distribution and spatial arrangement, more information is needed on the exact chemical composition of filler and matrix and their interaction with electromagnetic waves, including possible nonlinear effects.
Dental restorations should match the color of the surrounding enamel. Carefully selecting the appropriate shade for the filling material is a challenge for dentists. Moreover, tooth color can change over time due to habits such as smoking or drinking coffee. In the last few years, single-shade dental composites have come to the market. They rely on a chameleon effect to provide acceptable to good color matching regardless of the tooth color. The chameleon effect refers to a dental filling’s ability to guide light in such a way that its color blends in with that of the tooth. Structural color is a contributing factor to the chameleon effect and an active area of research where structures at the submicron scale play a critical role. We investigated the size, shape, and three-dimensional spatial arrangement of filler particles in single-shade dental resin composites. Cylindrical samples of dental composites were prepared and imaged with the transmission X-ray microscope at the ANATOMIX beamline, Synchrotron SOLEIL, France. The centers of the filler particles were determined from the tomography data. Combined with shape information from scanning electron microscopy, a Monte Carlo approach was used to model the transmittance for light at wavelengths from the visible to the ultraviolet. The results were compared to optical transmission measurements. The combination of nanotomography and simulation can thus help to understand the influence of the size and distribution of filler particles on the chameleon effect.
Cementum deposits on mammalian teeth contain layered microstructures associated with the chronological age of an animal and other details of their life history. Hard X-ray tomography data captured this record contained within the cementum deposits from whole teeth without sectioning. We investigated three teeth of African bovids, namely gemsbok (Oryx gazella), eland (Taurotragus oryx), and African or Cape buffalo (Syncerus caffer) using the laboratorybased system nanotom m for measuring each complete tooth to identify relevant regions, which were scanned at the ANATOMIX beamline of Synchrotron SOLEIL, France. Using microtomography in archaeological materials such as teeth, eliminates the need for tooth sectioning, making it a desirable alternative for archaeologists and museum curators. Synchrotron measurements enabled the application of pixel sizes as low as 0.65µm, which generated around 40 TB of data. The three adult bovids investigated here, have a known day of death and season of death, and come from regions with distinct seasonal patterns in temperature and/or rainfall. They also have an estimated age at death based on occlusal wear. The known information serves as a control to determine the applicability of microtomography on whole teeth of large bovids. Preliminary results show that microtomography can successfully replace the need of sectioning in cementum dental analysis. Our future goal is to develop a protocol to standardize procedures of tooth cementum analysis in bovids using microtomography.
Comparable to annual rings present in a tree trunk, human tooth cementum contains yearly deposited incremental layers often termed incremental lines, which are generally visualized from tooth slides with optical microscopy in two dimensions. These micrometer-thin incremental lines are used to decode age-at-death and stress periods over the lifetime of an individuum. One can also visualize these layers without physical slicing by means of hard X rays because of density modulations. Within this project, two optically almost transparent tooth slides were used to record optical data in two dimensions with submicron pixel sizes. These data were registered with projections of available synchrotron radiation-based tomography data of the slides. Such data were also acquired for an entire tooth to determine thickness variations in each layer, the intra-layer thickness, and variations between the layers, the inter-layer thickness, automatically.
Nanotechnology-based dental composites, see Tokuyama Dental's OMNICHROMA, can address the issue of color mismatch between tooth and filling. Similar to a chameleon, the filling can match the color of the surrounding enamel. We thoroughly investigated the nanostructure of the composite and the related optical properties using electron microscopy, synchrotron radiation-based nanotomography, small-angle X-ray scattering and optical transmission measurements. The spherical silica-zirconia fillers show a size of 260 nm and form micrometer-sized spherical domains of close-packed nanospheres. The aim of the study is to quantify the chameleon effect and discuss possible paths towards biomimetic anisotropic dental composites with improved color matching.
Hard X-ray micro computed tomography can be used for three-dimensional histological phenotyping of zebrafish embryos down to 1 µm or below without the need for staining or physical slicing. Current advances in ze- brafish embryo imaging, however, mostly rely on synchrotron radiation sources or highly advanced laboratory sources, which despite their evident strengths with regard to their beam properties and the implementation of phase contrast imaging techniques, lack accessibility. Therefore, we evaluated the performance of a conventional SkyScan 1275 laboratory µCT scanner in absorption contrast mode for the visualization of anatomical features in ethanol- and paraffin-embedded zebrafish embryos. We compare our results to readily available synchrotron data where 35 anatomical structures were identified. Despite having a more than ten times larger voxel length, approximately two thirds of the features could also be determined with laboratory microtomography. This could allow to monitor morphological changes during development or disease progression on large sample numbers, enabling the performance of preclinical studies in a local laboratory.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.