Sabrina Lang

Recent developments in X-ray Talbot interferometry at ESRF-ID19

In this paper we describe the design of different X-ray Talbot interferometers that have been built at the tomography beamline ID19 of the European Synchrotron Radiation Facility (ESRF) in Grenoble, France, and give a short review of performance characteristics, of current developments, and of the results obtained with these instruments so far. Among the applications so far, soft-tissue imaging has been a particular focus, as demonstrated in a recent paper by Schulz et al. (J. Roy. Soc. Interface, in press).

Experimental comparison of grating- and propagation-based hard X-ray phase tomography of soft tissue

When imaging soft tissues with hard X-rays, phase contrast is often preferred over conventional attenuation contrast due its superior sensitivity. However, it is unclear which of the numerous phase tomography methods yields the optimized results at given experimental conditions. Therefore, we quantitatively compared the three phase tomography methods implemented at the beamline ID19 of the European Synchrotron Radiation Facility: X-ray grating interferometry (XGI), and propagation-based phase tomography, i.e., single-distance phase retrieval (SDPR) and holotomography (HT), using cancerous tissue from a mouse model and an entire heart of a rat. We show that for both specimens, the spatial resolution derived from the characteristic morphological features is about a factor of two better for HT and SDPR compared to XGI, whereas the XGI data generally exhibit much better contrast-to-noise ratios for the anatomical features. Moreover, XGI excels in fidelity of the density measurements, and is also more robust against low-frequency artifacts than HT, but it might suffer from phase-wrapping artifacts. Thus, we can regard the three phase tomography methods discussed as complementary. The application will decide which spatial and density resolutions are desired, for the imaging task and dose requirements, and, in addition, the applicant must choose between the complexity of the experimental setup and the one of data processing.

Three-dimensional quantification of capillary networks in healthy and cancerous tissues of two mice

A key issue in developing strategies against diseases such as cancer is the analysis of the vessel tree in comparison to the healthy one. In the search for parameters that might be characteristic for tumor capillaries we study the vascularization in mice for cancerous and healthy tissues using synchrotron radiation-based micro computed tomography in absorption and phase contrast modes. Our investigations are based on absorption tomograms of casted healthy and cancerous tissues as well as a phase tomogram of a fixated tumor. We demonstrate how the voxel-based tomography data can be vectorized to assess the capillary networks quantitatively. The processing includes segmentation, skeletonization, and vectorization to finally extract the vessel parameters. The mean diameter of capillaries in healthy and cancerous tissues corresponds to (8.0±1.1) μm and (3.9±1.1) μm, respectively. Further evaluated parameters show marginal or no differences between capillaries in healthy and cancerous tissues, namely fractal dimension 2.3±0.3 vs. 2.3±0.2, tortuosity (SOAM) 0.18 rad/μm vs. 0.24 rad/μm and vessel length 20 μm vs. 17 μm. The bifurcation angles exhibit a narrow distribution around 115°. Furthermore, we show that phase tomography is a powerful alternative to absorption tomography of casts for the vessel visualization omitting any invasive specimen preparation procedure.

Holotomography versus X-ray grating interferometry: A comparative study

X-ray phase-contrast imaging techniques are used at synchrotron facilities to visualize tiny density variations in bulk samples. They overcome the limitations of other non-destructive methods, which often provide insufficient spatial and/or density resolution. Holotomography (HT) and X-ray grating interferometry (XGI) are among the most powerful phase-contrast techniques. Here, we show a direct comparison of HT versus XGI. We find that XGI excels in fidelity of the density measurements and is more robust against low-frequency artifacts, while HT is superior in spatial resolution. This study gives indications for applications and developments of phase-contrast imaging.

Three-dimensional registration of tomography data for quantification in biomaterials science

Abstract The paper shows the benefit of registration tools in the quantitative analysis of tomography data. For this purpose three-dimensional datasets, i.e. target design in rapid prototyping, hard X-ray tomography, and magnetic resonance tomography, were registered with rigid, affine and non-rigid algorithms. Using rigid registration, we aligned individual tomograms to obtain one three-dimensional dataset per specimen, compared human teeth to determine abrasion, and optimized the acquisition parameters in clinical computed tomography. Affine registration was used to determine the sintering-induced shrinkage of hydroxyapatite scaffolds. Using non-rigid registration, we gained insight into tumor morphology imaged by different modes of micro computed tomography. In addition, the fixation-induced shrinkage of tumor tissue and the local deformation of brain tissues as well as the shape changes in nickel titanium alloys were quantitatively determined. Three-dimensional registration is, therefore, a powerful means to count back any influence on morphology of tissues and to quantitatively compare data of different tomography modalities.

High-resolution tomographic imaging of microvessels

Cancer belongs to the primary diseases these days. Although different successful treatments including surgery, chemical, pharmacological, and radiation therapies are established, the aggressive proliferation of cancerous cells and the related formation of blood vessels has to be better understood to develop more powerful strategies against the different kinds of cancer. Angiogenesis is one of the crucial steps for the survival and metastasis formation of malignant tumors. Although therapeutic strategies attempting to inhibit these processes are being developed, the biological regulation is still unclear. This study concentrates on the three-dimensional morphology of vessels formed in a mouse tumor xenograft model post mortem. Synchrotron radiation-based micro computed tomography (SRμCT) could provide the necessary information that is essential for validating the simulations. Using mouse and human brain tissue, the different approaches to extract the vessel tree from SRμCT data are discussed. These approaches include corrosion casting, the application of contrast agents such as barium sulfate, tissue embedding, all of them regarded as materials science based. Alternatively, phase contrast tomography was used, which gave rise to promising results but still not reaches the spatial resolution to uncover the smallest capillaries.

Grating-based tomography of human tissues

The development of therapies to improve our health requires a detailed knowledge on the anatomy of soft tissues from the human body down to the cellular level. Grating-based phase contrast micro computed tomography using synchrotron radiation provides a sensitivity, which allows visualizing micrometer size anatomical features in soft tissue without applying any contrast agent. We show phase contrast tomography data of human brain, tumor vessels and constricted arteries from the beamline ID 19 (ESRF) and urethral tissue from the beamline W2 (HASYLAB/DESY) with micrometer resolution. Here, we demonstrate that anatomical features can be identified within brain tissue as well known from histology. Using human urethral tissue, the application of two photon energies is compared. Tumor vessels thicker than 20 μm can be perfectly segmented. The morphology of coronary arteries can be better extracted in formalin than after paraffin embedding.

Comparison of propagation-based phase-contrast tomography approaches for the evaluation of dentin microstructure

The complex hierarchical structure of human tooth hard tissues, enamel and dentin, guarantees function for decades. On the micrometer level the dentin morphology is dominated by the tubules, micrometer-narrow channels extending from the dentin-enamel junction to the pulp chamber. Their structure has been extensively studied, mainly with two-dimensional approaches. Dentin tubules are formed during tooth growth and their orientation is linked to the morphology of the nanometer-sized components, which is of interest for example for the development of bio-inspired dental fillings. Therefore, a method has to be identified that can access the three-dimensional organization of the tubules, e.g. density and orientation. Tomographic setups with pixel sizes in the sub-micrometer range allow for the three-dimensional visualization of tooth dentin tubules both in phase and absorption contrast modes. We compare high-resolution tomographic scans reconstructed with propagation based phase retrieval algorithms as well as reconstructions without phase retrieval concerning spatial and density resolution as well as rendering of the dentin microstructure to determine the approach best suited for dentin tubule imaging. Reasonable results were obtained with a single-distance phase retrieval algorithm and a propagation distance of about 75% of the critical distance of d2/λ, where d is the size of the smallest objects identifiable in the specimen and λ is the X-ray wavelength.

Global and local hard X-ray tomography of a centimeter-size tumor vessel tree

The visualization of the vascular network in tumors down to the smallest vessels requires high spatial resolution and reasonable contrast. Stained corrosion casts of the microvasculature network guarantee superior X-ray absorption contrast and highest reproduction fidelity. Tomography of a centimeter-size tumor, however, is unfeasible at the spatial resolution needed to reveal the smallest vessels. Therefore, local tomography has been performed to visualize the smallest capillaries within the region of interest. These three-dimensional data show the detailed morphology, but the reconstructed absorption coefficients obtained in local tomography differ substantially from the absorption coefficients retrieved from the less detailed global tomography data. This paper deals with the adaptation of local tomography data using the global data and considers two-parameter histogram matching of the radiographs, sinogram extension, and multi-parameter cupping correction. It is demonstrated that two-parameter histogram matching of the radiographs already provides reasonable agreement. The change of the lens in front of the detectors camera, however, significantly affects the obtained local X-ray absorption coefficients in the tomograms predominantly owing to the dissimilar point-spread functions of the two configurations used, and much less to the fact that one of the data sets was acquired in a local geometry.

Comparing the micro-vascular structure of cancerous and healthy tissues

Basic research is required to develop more powerful approaches to prevent, diagnose, and above all to treat cancer, although the clever combination of surgery, chemical, pharmacological, and radiation therapies is well established. Our research activity concentrates on the quantification of the three-dimensional micro-morphology of vessel trees formed in cancerous and healthy tissues of a mouse model post mortem. While in several cases it is possible to extract the vessel tree from corrosion casts, phase contrast imaging modalities are needed for cancerous tissues with a significant amount of damaged vessel walls. Differences between cancerous and healthy tissues could be identified. The sum-of-angle metrics is found to be constant for vessel segments in cancerous and healthy tissues with lengths between 12 and 220 μm and corresponds to (0.62 ± 0.10) rad/μm.