Jan Dudak

High-contrast X-ray micro-radiography and micro-CT of ex-vivo soft tissue murine organs utilizing ethanol fixation and large area photon-counting detector.

Using dedicated contrast agents high-quality X-ray imaging of soft tissue structures with isotropic micrometre resolution has become feasible. This technique is frequently titled as virtual histology as it allows production of slices of tissue without destroying the sample. The use of contrast agents is, however, often an irreversible time-consuming procedure and despite the non-destructive principle of X-ray imaging, the sample is usually no longer usable for other research methods. In this work we present the application of recently developed large-area photon counting detector for high resolution X-ray micro-radiography and micro-tomography of whole ex-vivo ethanol-preserved mouse organs. The photon counting detectors provide dark-current-free quantum-counting operation enabling acquisition of data with virtually unlimited contrast-to-noise ratio (CNR). Thanks to the very high CNR even ethanol-only preserved soft-tissue samples without addition of any contrast agent can be visualized in great detail. As ethanol preservation is one of the standard steps of tissue fixation for histology, the presented method can open a way for widespread use of micro-CT with all its advantages for routine 3D non-destructive soft-tissue visualisation.

Geometric correction methods for Timepix based large area detectors

X-ray micro radiography with the hybrid pixel detectors provides versatile tool for the object inspection in various fields of science. It has proven itself especially suitable for the samples with low intrinsic attenuation contrast (e.g. soft tissue in biology, plastics in material sciences, thin paint layers in cultural heritage, etc.). The limited size of single Medipix type detector (1.96 cm2) was recently overcome by the construction of large area detectors WidePIX assembled of Timepix chips equipped with edgeless silicon sensors. The largest already built device consists of 100 chips and provides fully sensitive area of 14.3 × 14.3 cm2 without any physical gaps between sensors. The pixel resolution of this device is 2560 × 2560 pixels (6.5 Mpix). The unique modular detector layout requires special processing of acquired data to avoid occurring image distortions. It is necessary to use several geometric compensations after standard corrections methods typical for this type of pixel detectors (i.e. flat-field, beam hardening correction). The proposed geometric compensations cover both concept features and particular detector assembly misalignment of individual chip rows of large area detectors based on Timepix assemblies. The former deals with larger border pixels in individual edgeless sensors and their behaviour while the latter grapple with shifts, tilts and steps between detector rows. The real position of all pixels is defined in Cartesian coordinate system and together with non-binary reliability mask it is used for the final image interpolation. The results of geometric corrections for test wire phantoms and paleo botanic material are presented in this article.

Development and characterization of high-resolution neutron pixel detectors based on Timepix read-out chips

Using a suitable isotope such as 6Li and 10B semiconductor hybrid pixel detectors can be successfully adapted for position sensitive detection of thermal and cold neutrons via conversion into energetic light ions. The adapted devices then typically provides spatial resolution at the level comparable to the pixel pitch (55 μm) and sensitive area of about few cm2. In this contribution, we describe further progress in neutron imaging performance based on the development of a large-area hybrid pixel detector providing practically continuous neutron sensitive area of 71 × 57 mm2. The measurements characterising the detector performance at the cold neutron imaging instrument ICON at PSI and high-flux imaging beam-line Neutrograph at ILL are presented. At both facilities, high-resolution high-contrast neutron radiography with the newly developed detector has been successfully applied for objects which imaging were previously difficult with hybrid pixel technology (such as various composite materials, objects of cultural heritage etc.). Further, a significant improvement in the spatial resolution of neutron radiography with hybrid semiconductor pixel detector based on the fast read-out Timepix-based detector is presented. The system is equipped with a thin planar 6LiF convertor operated effectively in the event-by-event mode enabling position sensitive detection with spatial resolution better than 10 μm.

Microtomography with photon counting detectors: improving the quality of tomographic reconstruction by voxel-space oversampling

Photon counting detectors Timepix are known for their unique properties enabling X-ray imaging with extremely high contrast-to-noise ratio. Their applicability has been recently further improved since a dedicated technique for assembling large area Timepix detector arrays was introduced. Despite the fact that the sensitive area of Timepix detectors has been significantly increased, the pixel pitch is kept unchanged (55 microns). This value is much larger compared to widely used and popular X-ray imaging cameras utilizing scintillation crystals and CCD-based read-out. On the other hand, photon counting detectors provide steeper point-spread function. Therefore, with given effective pixel size of an acquired radiography, Timepix detectors provide higher spatial resolution than X-ray cameras with scintillation-based devices unless the image is affected by penumbral blur. In this paper we take an advance of steep PSF of photon counting detectors and test the possibility to improve the quality of computed tomography reconstruction using finer sampling of reconstructed voxel space. The achieved results are presented in comparison with data acquired under the same conditions using a commercially available state-of-the-art CCD X-ray camera.

Evaluation of sample holders designed for long-lasting X-ray micro-tomographic scans of ex-vivo soft tissue samples

X-ray microradiography and microtomography are imaging techniques with increasing applicability in the field of biomedical and preclinical research. Application of hybrid pixel detector Timepix enables to obtain very high contrast of low attenuating materials such as soft biological tissue. However X-ray imaging of ex-vivo soft tissue samples is a difficult task due to its structural instability. Ex-vivo biological tissue is prone to fast drying-out which is connected with undesired changes of sample size and shape producing later on artefacts within the tomographic reconstruction. In this work we present the optimization of our Timepix equipped micro-CT system aiming to maintain soft tissue sample in stable condition. Thanks to the suggested approach higher contrast of tomographic reconstructions can be achieved while also large samples that require detector scanning can be easily measured.

Patterns of growth in monitor lizards (Varanidae) as revealed by computed tomography of femoral growth plates

Growth plate cartilage (GPC) is responsible for the growth of long bones due to endochondral ossification, which is the main mechanism of longitudinal skeletal growth in tetrapods. Degradation of GPC is a sign of determinate growth as it arrests the growth irreversibly. By contrast, indeterminate growth requires the persistence of GPC throughout the entire life. Monitor lizards (Varanidae) were previously reported to exhibit a dual type of growth, determinate as well as indeterminate. To reinvestigate this highly unexpected finding, we examined 13 species of varanids and their close relatives (Shinisauridae and Helodermatidae). In order to visualize GPC on the proximal part of the femur, we employed micro-radiography and micro-computed tomography. In large-bodied species, an extended capability of longitudinal growth was demonstrated; GPC was preserved for most of their lives. On the other hand, GPC senescence with complete disappearance of cartilage was found in adults of small-bodied varanids. These results confirm previous finding and, together with the absence of GPC in the outgroup species, challenge the universality of indeterminate growth in squamates. Moreover, we observed disappearance of GPC in an extremely old Varanus indicus, implying that GPC degradation is not entirely absent but only delayed to senescence in this large-bodied species. These findings raise the intriguing possibility that it is the timing, rather than other underlying mechanisms, what sets apart determinate from indeterminate growth. We therefore suggest that this dual type of growth represents an extreme case of heterochrony and is a consequence of strong sexual selection pressure to large-bodied varanids.

X-ray micro-CT and neutron CT as complementary imaging tools for non-destructive 3D imaging of rare silicified fossil plants

Computed tomography provides 3D information of inner structures of investigated objects. The obtained information is, however, strongly dependent on the used radiation type. It is known that as X-rays interact with electron cloud and neutrons with atomic nucleus, the obtained data often provide different contrast of sample structures. In this work we present a set of comparative radiographic and CT measurements of rare fossil plant samples using X-rays and thermal neutrons. The X-ray measurements were performed using large area photon counting detectors Timepix at IEAP CTU in Prague and Perkin Elmer flat-panel detector at Center of Excellence Telc. The neutron CT measurement was carried out at Paul Scherrer Institute using BOA beam-line. Furthermore, neutron radiography of fossil samples, provided by National Museum, were performed using a large-area Timepix detector with a neutron-sensitive converting 6LiF layer at Research Centre Rez, Czech Republic. The obtained results show different capabilities of both imaging approaches. While X-ray micro-CT provides very high resolution and enables visualization of fine cracks or small cavities in the samples neutron imaging provides high contrast of morphological structures of fossil plant samples, where X-ray imaging provides insufficient contrast.

X-ray imaging with sub-micron resolution using large-area photon counting detectors Timepix

As X-ray micro-CT became a popular tool for scientific purposes a number of commercially available CT systems have emerged on the market. Micro-CT systems have, therefore, become widely accessible and the number of research laboratories using them constantly increases. However, even when CT scans with spatial resolution of several micrometers can be performed routinely, data acquisition with sub-micron precision remains a complicated task. Issues come mostly from prolongation of the scan time inevitably connected with the use of nano-focus X-ray sources. Long exposure time increases the noise level in the CT projections. Furthermore, considering the sub-micron resolution even effects like source-spot drift, rotation stage wobble or thermal expansion become significant and can negatively affect the data. The use of dark-current free photon counting detectors as X-ray cameras for such applications can limit the issue of increased image noise in the data, however the mechanical stability of the whole system still remains a problem and has to be considered. In this work we evaluate the performance of a micro-CT system equipped with nano-focus X-ray tube and a large area photon counting detector Timepix for scans with effective pixel size bellow one micrometer.

Non-destructive imaging of fragments of historical beeswax seals using high-contrast X-ray micro-radiography and micro-tomography with large area photon-counting detector array.

Historical beeswax seals are unique cultural heritage objects. Unfortunately, a number of historical sealing waxes show a porous structure with a strong tendency to stratification and embrittlement, which makes these objects extremely prone to mechanical damage. The understanding of beeswax degradation processes therefore plays an important role in the preservation and consequent treatment of these objects. Conventional methods applied for the investigation of beeswax materials (e.g. gas chromatography) are of a destructive nature or bring only limited information about the sample surface (microscopic techniques). Considering practical limitations of conventional methods and ethical difficulties connected with the sampling of the historical material, radiation imaging methods such as X-ray micro-tomography presents a promising non-destructive tool for the onward scientific research in this field. In this contribution, we present the application of high-contrast X-ray micro-radiography and micro-tomography for the investigation of beeswax seal fragments. The method is based on the application of the large area photon-counting detector recently developed at our institute. The detector combines the advantages of single-photon counting technology with a large field of view. The method, consequently, enables imaging of relatively large objects with high geometrical magnification. In the reconstructed micro-tomographies of investigated historical beeswax seals, we are able to reveal morphological structures such as stratification, micro-cavities and micro-fractures with spatial resolution down to 5μm non-destructively and with high imaging quality. The presented work therefore demonstrates that a combination of state-of-the-art hybrid pixel semiconductor detectors and currently available micro-focus x-ray sources makes it possible to apply X-ray micro-radiography and micro-tomography as a valuable non-destructive tool for volumetric beeswax seal morphological studies.

High-contrast X-ray micro-tomography of low attenuation samples using large area hybrid semiconductor pixel detector array of 10 × 5 Timepix chips

State-of-the-art hybrid pixel semiconductor detectors provide excellent imaging properties such as unlimited dynamic range, high spatial resolution, high frame rate and energy sensitivity. Nevertheless, a limitation in the use of these devices for imaging has been the small sensitive area of a few square centimetres. In the field of microtomography we make use of a large area pixel detector assembled from 50 Timepix edgeless chips providing fully sensitive area of 14.3 × 7.15 cm2. We have successfully demonstrated that the enlargement of the sensitive area enables high-quality tomographic measurements of whole objects with high geometrical magnification without any significant degradation in resulting reconstructions related to the chip tilling and edgeless sensor technology properties. The technique of micro-tomography with the newly developed large area detector is applied for samples formed by low attenuation, low contrast materials such a seed from Phacelia tanacetifolia, a charcoalified wood sample and a beeswax seal sample.