Synchrotron imaging of bovine and human ovaries ex vivo

Background and Rationale: Reproductive dysfunction affects more than 15% of Canadian women; however, the underlying causes remain largely unknown. Ultrasonography is the most commonly used research and diagnostic tool for imaging the ovaries and uterus. However, current ultrasonographic techniques allow the detection of ovarian structures (eg. follicles, corpora lutea) at diameters of only ≥2 mm. The increased effectiveness of synchrotron technology for imaging ovaries in comparison to conventional imaging methods is currently unknown. Overall Objective: The overall objective of this research was to determine the effectiveness of synchrotron techniques for imaging ovaries. We hypothesized that synchrotron techniques would provide greater contrast for visualizing structural details of follicles, corpora lutea (CL), and cumulus oocyte complexes (COC), compared to conventional ultrasonography. Materials and Methods: Three studies were conducted to evaluate phase-contrast based synchrotron imaging methods. The first study involved Diffraction Enhanced Imaging (DEI) of bovine ovaries (n=6). The second study involved Propagation-Based Computed Tomography (PB-CT) imaging of bovine (n=4) and human ovaries (n=4). A third, preliminary study was conducted to explore the use of Talbot Grating Interferometry (TGI-CT) imaging of bovine (n=1) and human ovaries (n=1). Fresh and formalin-fixed bovine and human ovaries were imaged without or with contrast injection into the ovarian artery. Following synchrotron imaging, all ovarian samples were evaluated using diagnostic ultrasonography and histology. Images obtained using synchrotron techniques, ultrasonography and histology were qualitative and quantitatively compared. Results: DEI allowed the identification of 71% of follicles ≥2 mm and 67% of CL detected using ultrasonography. Mean follicle diameter was similar between DEI (9.6 ± 2.4 mm), ultrasonography (9.0 ± 2.6 mm), and histology (6.9 ± 1.9 mm) for fresh ovaries without contrast (P = 0.70). Likewise, no difference in CL diameter was detected between DEI (11.64 ± 1.67 mm), ultrasonography (9.34 ± 0.35 mm), and histology (9.6 ± 0.4 mm), (P = 0.34). Antral Follicle Count (AFC; ≥2mm) was similar between ultrasonography (6.5 ± 0.7 mm, fresh with no contrast; 6.5 ± 2.5 mm, preserved with no contrast) and DEI ( 4.5 ± 0.5 mm, fresh with no contrast; 6.5 ± 0.50 mm, preserved with no contrast) (P > 0.05). However, the contrast resolution for differentiating follicles and CL was inferior with DEI compared to ultrasonography. Small antral follicles <2mm, cell layers comprising the follicle wall and COC were not detected using either DEI or ultrasonography. PB-CT imaging enabled the visualization of 100% of follicles ≥2 mm and 100% of CL that were detected with ultrasonography. CL containing a central cystic cavity were identified using PB-CT; however, CL without a central cystic cavity were not well-visualized. Mean follicle and luteal diameters did not differ among PB-CT, ultrasonography and histology (P>0.05). PB-CT was superior to ultrasonography for detecting small antral follicles <2 mm in bovine ovaries (P = 0.04), and the granulosa and theca cell layers of the follicle wall in bovine and human ovaries (P < 0.0001). However, TGI-CT images exhibited greater contrast resolution for visualizing small and large antral follicles, CL, and the cell layers of the follicle wall compared to both PB-CT and ultrasonography. High contrast structures resembling COC were detected with both PB-CT and TGI-CT, but not with ultrasonography. Only TGI-CT permitted the visualization of the oocyte within the COC in fresh and preserved ovaries. Conclusions: DEI was inferior to ultrasonography for detecting ovarian follicles and CL. PB-CT was superior to ultrasonography for visualizing follicles <2 mm, COC, and the cell layers of the follicle wall. However, PB-CT was as effective as ultrasonography for detecting and measuring follicles ≥2 mm and cystic CL. Preliminary findings suggest that TGI-CT provides the greatest contrast for imaging both ovarian macro- and microanatomy compared to PB-CT, DEI, and ultrasonography

Development of X-ray phase-contrast imaging techniques for medical diagnostics

The X-Ray phase-contrast techniques are innovative imaging methods allowing overtaking the limitations of classic radiology. In addition to the differential X-ray absorption on which standard radiology relies, in phase-contrast imaging the contrast is given by the effects of the refraction of X-rays inside the tissues. The combination of phase-contrast with quantitative computer tomography (CT) allows for a highly accurate reconstruction of the tissues’ index of refraction. Thanks to the high sensitivity of the method, tomographic images can be obtained at clinically compatible dose. For all these reasons phase-contrast imaging is a very promising approach, which can potentially revolutionize diagnostic X-Ray imaging. Several techniques are classified under the name of X-Ray phase-contrast imaging. This Thesis focused on the so-called analyzer-based imaging (ABI) method. ABI uses a perfect crystal, placed between the sample and the detector, to visualize the phase effects occurred within the sample. The quantitative reconstruction of the refraction index from CT data is not trivial and before this Thesis work it was documented only for small size objects. This Thesis has focused on two main scientific problems: (1) the development of theoretical and calculation strategies to determine the quantitative map of the refraction index of large biological tissues/organs (>10 cm) using the ABI technique; and (2) the preparation of accurate and efficient tools to estimate and simulate the dose deposited in CT imaging of large samples. For the determination of the refraction index, two CT geometries were considered and studied: the out-of-plane and the in-plane configurations. The first one, the most used in the works reported in the literature, foresees that the rotation axis of the sample occurs in a plane parallel to that of the sensitivity of the analyzer crystal; while, in the second CT geometry, the rotation axis is perpendicular to that plane. The theoretical study, technical design and experimental implementation of the in-plane geometry have been main tasks of this Thesis. A first experiment has been performed in order to compare the results obtained with in-plane quantitative phase contrast CT with the absorption-based CT ones. An improved accuracy and a better agreement with the theoretical density values have been obtained by exploiting the refraction effect while keeping the dose to sample low. A second campaign of experiments has been performed on large human breasts to investigate the efficiency of the in-plane and out-of-plane CT geometries and the performances of the associated image reconstruction procedures. The same experimental conditions were also studied by numerical simulations and the results were compared. This analysis shows that the in-plane geometry allows producing more accurate quantitative three dimensional maps of the index of refraction, while the out-of-plane case is preferable for qualitative investigations. A study for developing advanced procedures for improving the quality of the obtained CT images has been also conducted. As a result, a two-step procedure has been tested and identified: first the noise level of the experimental images is reduced by applying a wavelet decomposition algorithm and then a deconvolution procedure. The obtained images show an enhanced sharpness of the interfaces and of the object edges and high signal to noise ratio values are preserved. The second problem of this Thesis was to find strategies to calculate, in a fast way, the delivered dose in CT imaging of complex biological samples. For this purpose an acceleration method to speed-up the convergence of Monte Carlo simulations based on the Track Length Estimator method has been computed and included in the open-source software GATE. Results show that this method can lead to the same accuracy of conventional Monte Carlo methods while reducing the required computation time of up to two orders of magnitude, with the respect to the considered geometry. A database of dose curves for the case of monochromatic breast CT has been produced: it allows for a quick estimation of the delivered dose. A way to choose the best energy and the optimal photon flux was also proposed, which leads to a significant reduction of the delivered dose without any loss in terms of image quality. Most of the experimental and data reconstruction methods developed within this Thesis work can be applied also to other phase-contrast techniques. This Thesis shows that high resolution three dimensional diagnostic imaging of large and complex biological organs can, in principle, be performed at clinical compatible doses; this is the most significant contribution of the Thesis towards the clinical implementation of phase-contrast CT.Auf Phasenkontrast basierende Röntgentechniken sind innovative bildgebende Methoden, welche die Limitierungen der klassischen Radiologie überschreiten. Auβer der differentiellen Röntgenabsorption, auf der die herkömmliche Radiologie beruht, ist der Kontrast bei Phasenkontrast-Bildgebung durch die Brechungseffekte der Röntgenstrahlen innerhalb eines Gewebes gegeben. Die Kombination zwischen Phasenkontrast und quantitativer Computertomographie (CT) erlaubt eine höchstgenaue Rekonstruktion der Brechzahl der Gewebe. Aufgrund der hohen Empfindlichkeit dieser Methode, können tomographische Bilder mit einer klinisch verträglichen Dosis erzeugt werden. Aus all diesen Gründen, stellt Phasenkontrast-Bildgebung einen vielversprechenden Ansatz dar, welcher die diagnostische Röntgenbildgebung revolutionieren könnte. Verschiedene röntgenbildgebende Techniken werden als Phasenkontrast-Verfahren bezeichnet. Die vorliegende Doktorarbeit befasst sich mit der sogenannten Bildgebungsmethode mithilfe eines Analysatorkristalls (auf englisch: analyser-based imaging (ABI) ). ABI benutzt ein perfektes, zwischen der Probe und dem Detektor angeordnetes Kristall, um in der Probe stattfindenden Phaseneffekte zu veranschaulichen. Die quantitative Rekonstruktion des Brechungsindizes aus den CT-Daten ist jedoch nicht trivial und war vor dieser Arbeit nur für kleine Gegenstände beschrieben. Im Mittelpunkt dieser Dissertation stehen folgende wissenschaftliche Fragestellungen: (1) die Entwicklung theoretischer und rechnerischer Strategien, um die quantitative räumliche Verteilung des Brechungsindizes in größeren Organen aus biologischen Geweben (10 cm) unter Verwendung der ABI-Technik zu bilden und (2) die Vorbereitung von genauen und leistungsfähigen Rechenmitteln zur Abschätzung und Simulation der in größeren Proben bei einem CT-Bildgebungsversuch abgelagerten Strahlendosis zu treffen. Für die Bestimmung des Brechungsindizes wurden zwei geometrische Anordnungen in Betracht gezogen und untersucht, und zwar die Konfiguration auβerhalb (out-of-plane) bzw. in der Ebene (in-plane) der Probe. Erstere wird am häufigsten in der Fachliteratur zitiert und sieht vor, dass die Probe-Drehachse sich in der parallelen Ebene zur Achse des Analysatorkristalls befindet, wobei in der zweiteren Geometrie die Drehachse orthogonal zu jener Ebene ist. Die theoretische Studie, der technische Entwurf und die experimentelle Umsetzung der geometrischen Anordnung in der Probe-Ebene stellen die Hauptaufgaben dieser Arbeit dar. Ein erstes Experiment wurde durchgeführt, um die durch quantitative Phasenkontrast-CT nach in-plane-Modus erlangten Ergebnisse mit entsprechenden, auf Absorption basierenden CT-Versuchen zu vergleichen. Eine höhere Genauigkeit sowie eine bessere Übereinstimmung mit den theoretischen Dichtewerten wurden dadurch erzielt, dass man sich die Brechungseffekte zunutze macht, indem man die an die Probe gelieferte Dosis niedrig hält. Eine zweite Versuchsreihe wurde auβerdem auf menschliche Brüste ausgeführt, um die Effizienz sowohl der in-plane- als auch der out-of-plane-CT-Geometrien sowie die Leistungsfähigeit der entsprechenden Bildrekonstruktionsverfahren zu überprüfen. Die gleichen Experimentalbedingungen wurden auch anhand von numerischen Simulationen untersucht und die Ergebnisse miteinander verglichen. Diese Analyse zeigt, dass die in-plane-Geometrie die Erstellung genauerer dreidimensionaler Verteilungen der Brechzahl ermöglicht, während der out-of-plane-Fall eher für die Zwecke qualitativer Untersuchungen vorzuziehen ist. Fortschrittliche Prozeduren zur Verbesserung der Qualität von aufgezeichneten CT-Bildern wurden im Rahmen dieser Doktorarbeit konzipiert und entwickelt. Das Fazit: eine zweistufige Vorgehensweise wurde ermittelt und geprüft. Zunächst wird der Rauschpegel der Meβdaten über die Anwendung eines Zerlegungsalgorithmus mittels Wavelets gesenkt, anschlieβend gefolgt von einem Entfaltung-Verfahren. Die damit gewonnenen Bilder weisen eine erhöhte Schärfe der Schnittstellen auf. Die Objektkanten und das Signal-zu-Rausch-Verhältnis bleiben damit erhalten. Die zweite Fragestellung dieser Arbeit war es, Lösungansätze zu erarbeiten, um die während CT-Bildgebung-Messungen über complexe biologische Proben abgegebene Dosis möglichst rapide zu berechnen. Zu diesem Zweck wurde ein Verfahren zur Beschleunigung der Konvergenz von Monte-Carlo-Simulationen auf der Grundlage der Track-Length-Estimator-Methode entwickelt und in die Open-Source-Software GATE eingegliedert. Die bisherigen Ergebnisse zeigen, dass dieses Verfahren zur selben Genauigkeit der herkömmlichen Monte-Carlo-Methoden bei gleichzeitiger Minderung bis zu zwei Gröβenordnungen der zur Berechnung einer und der selben Geometrie notwendigen Rechenzeit führt. Eine Datenbank von Dosiskurven für den Fall von monochromatischer Brust-CT ist erzeugt worden, die eine schnelle Schätzung der abgegebenen Dosis erlaubt. Darüber hinaus wurde ein Lösungsweg zur Auswahl der besten Energie und des optimalen Photonenflusses vorgeschlagen, welcher eine bedeutende Abnahme der abgelieferten Dosis zur Folge hat, und zwar ohne Bildqualitätsverluste. Die meisten, im Rahmen dieser Doktorarbeit entwickelten Experimental- und Datenrekonstruktion-Verfahren können freilich auch an andere Phasenkontrast-Techniken angewendet werden. Es wird hiermit gezeigt, dass hochauflösende dreidimensionale bildgebende Verfahren zur Diagnostik gröβerer und komplexer biologischer Gegenstände bei klinisch verträglichen Dosen grundsätzlich eingesetzt werden können. Dies ist der nennenwerteste Beitrag dieser Dissertation zur klinischen Umsetzung der Phasenkontrast-CT

DEVELOPMENT AND CHARACTERIZATION OF A HIGH-ENERGY IN-LINE PHASE CONTRAST TOMOSYNTHESIS PROTOTYPE

Phase sensitive 3D imaging techniques have been an emerging field in x-ray imaging for two decades. Among them, in-line phase contrast tomosynthesis has been investigated with great potential for translation into clinical applications in the near future, due to combining the advantages of configuration simplicity, structural noise elimination and potentially low radiation dose delivery. The high-energy in-line phase contrast tomosynthesis technique developed and presented in this dissertation initiates this translational procedure by optimizing the imaging conditions, performing phase retrieval, offering opportunities to further reduce radiation dose delivery, improving detectability and specificity with the employment of auxiliary phase contrast agents, and potentially performing quantitative imaging. First, the high-energy in-line phase contrast tomosynthesis prototype was developed and characterized in this dissertation as the first of its kind following a number of engineering trade-off considerations. The quantitative results as well as the imaging results of tissue-simulating phantoms and biology-related phantoms demonstrate the extensive capability of this imaging prototype in improving tumor detectability. In addition, the optimization of the x-ray prime beam toward the PAD phase retrieval method proved the potential of high-energy imaging and predicated the solution toward imaging time reduction by employing photon counting based imaging techniques. In the past several years, applications of microbubbles as a phase contrast agent have shown the capability for image quality improvement in quantitative imaging. In this dissertation, a preliminary study of quantitative imaging of microbubbles using the in-line phase contrast projection mode imaging prototype, which is a system without tomosynthesis capability, provided a discussion on how the materials of the bubble shells and gas infills could impact the imaging capabilities and resulting image detectability. In addition, the results of the study provided a guideline for microbubble selections for in-line phase contrast mode imaging modalities. Based on this criterion discussed in the study, the albumin-shell microbubbles were selected as the phase contrast agent for the imaging prototype presented in this dissertation. The imaging results showed the feasibility of performing quantitative imaging by employing microbubbles as the auxiliary phase contrast agent. Clinical conditions were simulated by distributing microbubbles on the interface between two tissue-like phantom structures. The quantitative imaging results provided clinical motivation for translating phantom studies into more biology-related investigations providing radiation dose reductions in the future

A comparison of different approaches to image quality assessment in phase-contrast mammography

Introduction: The purpose of medical imaging in breast cancer screening is to detect and characterise pathology. In this context, image quality is best defined in relation to diagnostic performance. However, in many situations, the only practical and economical method to assess image quality is to use clinical image quality assessment performed by radiologists. Clinical image quality assessment can be expensive, time-consuming and suffers from intra and inter-observer variability. Therefore, it is useful to establish a robust, quantitative image assessment method that can accurately predict radiologists’ clinical image quality assessment. Unfortunately, the variable anatomical backgrounds in clinical images significantly complicate the relationship between physical and clinical image qual- ity. Two-dimensional digital X-ray mammography (DM) is currently the most commonly utilised screening modality in breast cancer screening. However, it has well-documented limitations. Promising alternatives to two-dimensional DM involving phase-contrast X-ray imaging are currently under investigation. Phase-contrast imaging is an Xray imaging technique where image contrast is not only related to the X-ray attenuation properties of tissue (as is the case with conventional radiography) but also the refractive properties of the tissue. One technique currently under active research at synchrotron facilities is propagation-based phase-contrast computed tomography (PB-CT). In this work, we take advantage of the relatively simple anatomical background present in synchrotron-based, thin slice PB-CT images of breasts to formulate a simple, robust relative image assessment model. Methods: The experimental data analysed in this study included PB-CT scans, which were obtained for twelve whole, intact mastectomy samples at Imaging and Medical beam- line (IMBL) of the Australian Synchrotron. Eleven radiologists assessed overall clinical image quality. Physical image metrics, including contrast, signal to noise ratio, and spatial resolution, were calculated using two different methods for all PB-CT and conventional CT image sets. Weighting factors were applied to each metric, and a scaled contrast to noise (CNR) to spatial resolution (res) score (CNR/res) was calculated. Results: The scaled CNR/res score for each imaging condition, averaged across all samples, was found to correlate significantly with the corresponding radiologist scores with a Pearson r value of approximately 0.96. In addition, the CNR/res score for each imaging condition, for each sample, also correlated significantly with the corresponding radiologist scores with a Spearman r value of approximately 0.89. Conclusion: The scaled CNR/res criterion has been demonstrated as a quantitative image assessment model that effectively predicts the relative clinical image quality, as assessed by radiologists, in the context of PB-CT breast imaging

Comparison of Unmonochromatized Synchrotron Radiation and Conventional X-rays in the Imaging of Mammographic Phantom and Human Breast Specimens: A Preliminary Result

A simple imaging setup based on the principle of coherence-based contrast X-ray imaging with unmonochromatized synchrotron radiation was used for studying mammographic phantom and human breast specimens. The use of unmonochromatized synchrotron radiation simplifies the instrumentation, decreases the cost and makes the procedure simpler and potentially more suitable for clinical applications. The imaging systems consisted of changeable silicon wafer attenuators, a tungsten slit system, a CdWO4 scintillator screen, a CCD (Charge Coupled Device) camera coupled to optical magnification lenses, and a personal computer. In preliminary studies, a spatial resolution test pattern and glass capillary filled with air bubbles were imaged to evaluate the resOolution characteristics and coherence-based contrast enhancement. Both the spatial resolution and image quality of the proposed system were compared with those of a conventional mammography system in order to establish the characteristic advantages of this approach. The images obtained with the proposed system showed a resolution of at least 25 µm on the test pattern with much better contrast, while the images of the capillary filled with air bubbles revealed coherence-based edge enhancement. This result shows that the coherence-based contrast imaging system, which emphasizes the refraction effect from the edge of materials of different refractive indexes, is applicable to imaging studies in fundamental medicine and biology, although further research works will be required before it can be used for clinical applications.ope

Evaluation of a diffraction-enhanced imaging (DEI) prototype and exploration of novel applications for clinical implementation of DEI

Conventional mammographic image contrast is derived from x-ray absorption, resulting in breast structure visualization due to density gradients that attenuate radiation without distinction between transmitted, scattered, or refracted x-rays. Diffraction-enhanced imaging (DEI) allows for increased contrast with decreased radiation dose compared to conventional mammographic imaging due to monochromatic x-rays, its unique refraction-based contrast mechanism, and excellent scatter rejection. Although laboratory breast imaging studies have demonstrated excellent breast imaging, important clinical translation and application studies are needed before the DEI system can be established as a useful breast imaging modality. This dissertation focuses on several important studies toward the development of a clinical DEI system. First, contrast-enhanced DEI was explored using commercially available contrast agents. Phantoms were imaged at a range of x-ray energies and relevant contrast agent concentrations. Second, we performed a reader study to determine if superior DEI contrast mechanisms preserve image quality as tissue thickness increases. Breast specimens were imaged at several thicknesses, and radiologist perception of lesion visibility was recorded. Lastly, a prototype DEI system utilizing an x-ray tube source was evaluated through a reader study. Breast tissue specimens were imaged on the traditional and prototype DEI systems, and expert radiologists evaluated image quality and pathology correlation. This dissertation will demonstrate proof-of-principle for contrast-enhanced DEI, establishing the feasibility of contrast-enhanced DEI using commercially available contrast agents. Further, it will show that DEI might be able to reduce breast compression, and thus the perception of pain during mammography, without significantly decreasing breast lesion visibility. Finally, this research shows the successful implementation of a DEI prototype, displaying breast features with approximately statistically equivalent visibility to the traditional DEI system. Together, this research is an important step toward the clinical translation of DEI, a technology with the potential to facilitate early breast cancer detection and diagnosis

System parameters and performance specifications for the application of Diffraction Enhanced Imaging and Multiple Image Radiography to breast imaging

The Diffraction Enhanced Imaging (DEI) method is a novel x-ray imaging technique that dramatically extends the capability of conventional x-ray imaging. X-ray imaging has traditionally been dependent on x-ray absorption to generate contrast, and is the physical mechanism of contrast in planar x-ray imaging and computed tomography. DEI utilizes the Bragg peak of perfect crystal diffraction to convert angular changes into intensity changes, providing a large change in intensity for a small change in angle. The use of a silicon analyzer crystal in the path of the x-ray beam generates two additional forms of image contrast, refraction and extinction. Objects that have very little absorption contrast may have considerable refraction and extinction contrast, this improving visualization and extending the utility of x-ray imaging. An area of medicine where this technique could have a dramatic impact is in breast imaging, where the key diagnostic structures often have low absorption contrast, especially in the early stages of disease. In order to develop a DEI clinical prototype imaging system, a systematic assessment of the engineering parameters for the breast imaging application must be determined. This body of work investigates the primary imaging parameters of DEI (x-ray beam energy, crystal reflections, angular sampling) and demonstrates how the unique properties of DEI can be capitalized upon to address the engineering limitations of flux, dramatically reducing the dose required for imaging. The results from this analysis are used to describe a plausible design for a non-synchrotron based DEI breast imaging system