Reconstructed tomographic image resolution of small animal PET imaging systems is improving with advances in radiation detector development. However the trend towards higher resolution systems has come with an increase in price and system complexity. Recent developments in the area of solid-state photomultiplication devices like silicon photomultiplier arrays (SPMA) are creating opportunities for new high performance tools for PET scanner design. Imaging of excised small animal organs and tissues has been used as part of post-mortem studies in order to gain detailed, high-resolution anatomical information on sacrificed animals. However, this kind of ex-vivo specimen imaging has largely been limited to ultra-high resolution μCT. The inherent limitations to PET resolution have, to date, excluded PET imaging from these ex-vivo imaging studies. In this work, we leverage the diminishing physical size of current generation SPMA designs to create a very small, simple, and high-resolution prototype detector system targeting ex-vivo tomographic imaging of small animal organs and tissues. We investigate sensitivity, spatial resolution, and the reconstructed image quality of a prototype small animal PET scanner designed specifically for imaging of excised murine tissue and organs. We aim to demonstrate that a cost-effective silicon photomultiplier (SiPM) array based design with thin crystals (2 mm) to minimize depth of interaction errors might be able to achieve sub-millimeter resolution. We hypothesize that the substantial decrease in sensitivity associated with the thin crystals can be compensated for with increased solid angle detection, longer acquisitions, higher activity and wider acceptance energy windows (due to minimal scatter from excised organs). The constructed system has a functional field of view (FoV) of 40 mm diameter, which is adequate for most small animal specimen studies. We perform both analytical (3D-FBP) and iterative (ML-EM) methods in order to reconstruct tomographic images. Results demonstrate good agreement between the simulation and the prototype. Our detector system with pixelated crystals is able to separate small objects as close as 1.25 mm apart, whereas spatial resolution converges to the theoretical limit of 1.6 mm (half the size of the smallest detecting element), which is to comparable to the spatial resolution of the existing commercial small animal PET systems. Better system spatial resolution is achievable with new generation SiPM detector boards with 1 mm x 1 mm cell dimensions. We demonstrate through Monte Carlo simulations that it is possible to achieve sub-millimeter spatial image resolution (0.7 mm for our scanner) in complex objects using monolithic crystals and exploiting the light-sharing mechanism among the neighboring detector cells. Results also suggest that scanner (or object) rotation minimizes artifacts arising from poor angular sampling, which is even more significant in smaller PET designs as the gaps between the sensitive regions of the detector have a more exaggerated effect on the overall reconstructed image quality when the design is more compact. Sensitivity of the system, on the other hand, can be doubled by adding two additional detector heads resulting in a, fully closed, 4π geometry.

Herein we present the development of a fluorescence-enhanced, frequency-domain radiative transport reconstruction system designed for small animal optical tomography. The system includes a time-dependent data acquisition instrument, a radiative transport based forward model for prediction of time-dependent propagation of photons in small, non-diffuse volumes, and an algorithm which utilizes the forward model to reconstruct fluorescent yields from air/tissue boundary measurements. The major components of the instrumentation include a charge coupled device camera, an image intensifier, signal generators, and an optical switch. Time-dependent data were obtained in the frequency-domain using homodyne techniques on phantoms with 0.2% to 3% intralipid solutions. Through collaboration with Transpire, Inc., a fluorescence-enhanced, frequency-domain, radiative transport equation (RTE) solver was developed. This solver incorporates the discrete ordinates, source iteration with diffusion synthetic acceleration, and linear discontinuous finite element differencing schemes, to predict accurately the fluence of excitation and emission photons in diffuse and transport limited systems. Additional techniques such as the first scattered distributed source method and integral transport theory are used to model the numerical apertures of fiber optic sources and detectors. The accuracy of the RTE solver was validated against diffusion and Monte Carlo predictions and experimental data. The comparisons were favorable in both the diffusion and transport limits, with average errors of the RTE predictions, as compared to experimental data, typically being less than 8% in amplitude and 7% in phase. These average errors are similar to those of the Monte Carlo and diffusion predictions. Synthetic data from a virtual mouse were used to demonstrate the feasibility of using the RTE solver for reconstructing fluorescent heterogeneities in small, non-diffuse volumes. The current version of the RTE solver limits the reconstruction to one iteration and the reconstruction of marginally diffuse, frequency-domain experimental data using RTE was not successful. Multiple iterations using a diffusion solver successfully reconstructed the fluorescent heterogeneities, indicating that, when available, multiple iterations of the RTE based solver should also reconstruct the heterogeneities.

<p>X-ray micro-CT is widely used for small animal imaging in preclinical studies of cardiopulmonary disease, but further development is needed to improve spatial resolution, temporal resolution, and material contrast. This study presents a set of tools that achieve these improvements. These tools include the mathematical formulation and computational implementation of algorithms for calibration, image reconstruction, and image analysis with our custom micro-CT system. These tools are tested in simulations and in experiments with live animals. With these tools, it is possible to visualize the distribution of a contrast agent throughout the body of a mouse as it changes over time, and produce 5-dimensional images (3 spatial dimensions + time + energy) of the cardiac cycle.</p> / Dissertation

Herein we present the development of a fluorescence-enhanced, frequency-domain radiative transport reconstruction system designed for small animal optical tomography. The system includes a time-dependent data acquisition instrument, a radiative transport based forward model for prediction of time-dependent propagation of photons in small, non-diffuse volumes, and an algorithm which utilizes the forward model to reconstruct fluorescent yields from air/tissue boundary measurements. The major components of the instrumentation include a charge coupled device camera, an image intensifier, signal generators, and an optical switch. Time-dependent data were obtained in the frequency-domain using homodyne techniques on phantoms with 0.2% to 3% intralipid solutions. Through collaboration with Transpire, Inc., a fluorescence-enhanced, frequency-domain, radiative transport equation (RTE) solver was developed. This solver incorporates the discrete ordinates, source iteration with diffusion synthetic acceleration, and linear discontinuous finite element differencing schemes, to predict accurately the fluence of excitation and emission photons in diffuse and transport limited systems. Additional techniques such as the first scattered distributed source method and integral transport theory are used to model the numerical apertures of fiber optic sources and detectors. The accuracy of the RTE solver was validated against diffusion and Monte Carlo predictions and experimental data. The comparisons were favorable in both the diffusion and transport limits, with average errors of the RTE predictions, as compared to experimental data, typically being less than 8% in amplitude and 7% in phase. These average errors are similar to those of the Monte Carlo and diffusion predictions. Synthetic data from a virtual mouse were used to demonstrate the feasibility of using the RTE solver for reconstructing fluorescent heterogeneities in small, non-diffuse volumes. The current version of the RTE solver limits the reconstruction to one iteration and the reconstruction of marginally diffuse, frequency-domain experimental data using RTE was not successful. Multiple iterations using a diffusion solver successfully reconstructed the fluorescent heterogeneities, indicating that, when available, multiple iterations of the RTE based solver should also reconstruct the heterogeneities.

The images in positron emission tomography (PET) represent three dimensional dynamic distributions of biologically interesting molecules labelled with positron emitting radionuclides (radiotracers). Spatial localisation of the radio-tracers is achieved by detecting in coincidence two collinear photons which are emitted when the positron annihilates with an ordinary electron. In order to obtain quantitatively accurate images in PET, it is necessary to correct for the effects of photon attenuation within the subject being imaged. These corrections can be obtained using singles-mode photon transmission scanning. Although suitable for small animal PET, these scans are subject to high amounts of contamination from scattered photons. Currently, no accurate correction exists to account for scatter in these data. The primary purpose of this work was to implement and validate an analytical scatter correction for PET transmission scanning. In order to isolate the effects of scatter, we developed a simulation tool which was validated using experimental transmission data. We then presented an analytical scatter correction for singles-mode transmission data in PET. We compared our scatter correction data with the previously validated simulation data for uniform and non-uniform phantoms and for two different transmission source radionuclides. Our scatter calculation correctly predicted the contribution from scattered photons to the simulated data for all phantoms and both transmission sources. We then applied our scatter correction as part of an iterative reconstruction algorithm for simulated and experimental PET transmission data for uniform and non-uniform phantoms. We also tested our reconstruction and scatter correction procedure using transmission data for several animal studies (mice, rats and primates). For all studies considered, we found that the average reconstructed linear attenuation coefficients for water or soft-tissue regions of interest agreed with expected values to within 4%. Using a 2.2 GHz processor, the scatter correction required between 6 to 27 minutes of CPU time (without any code optimisation) depending on the phantom size and source used. This extra calculation time does not seem unreasonable considering that, without scatter corrections, errors in the reconstructed attenuation coefficients were between 18 to 45% depending on the phantom size and transmission source used.

The images in positron emission tomography (PET) represent three dimensional dynamic distributions of biologically interesting molecules labelled with positron emitting radionuclides (radiotracers). Spatial localisation of the radio-tracers is achieved by detecting in coincidence two collinear photons which are emitted when the positron annihilates with an ordinary electron. In order to obtain quantitatively accurate images in PET, it is necessary to correct for the effects of photon attenuation within the subject being imaged. These corrections can be obtained using singles-mode photon transmission scanning. Although suitable for small animal PET, these scans are subject to high amounts of contamination from scattered photons. Currently, no accurate correction exists to account for scatter in these data. The primary purpose of this work was to implement and validate an analytical scatter correction for PET transmission scanning. In order to isolate the effects of scatter, we developed a simulation tool which was validated using experimental transmission data. We then presented an analytical scatter correction for singles-mode transmission data in PET. We compared our scatter correction data with the previously validated simulation data for uniform and non-uniform phantoms and for two different transmission source radionuclides. Our scatter calculation correctly predicted the contribution from scattered photons to the simulated data for all phantoms and both transmission sources. We then applied our scatter correction as part of an iterative reconstruction algorithm for simulated and experimental PET transmission data for uniform and non-uniform phantoms. We also tested our reconstruction and scatter correction procedure using transmission data for several animal studies (mice, rats and primates). For all studies considered, we found that the average reconstructed linear attenuation coefficients for water or soft-tissue regions of interest agreed with expected values to within 4%. Using a 2.2 GHz processor, the scatter correction required between 6 to 27 minutes of CPU time (without any code optimisation) depending on the phantom size and source used. This extra calculation time does not seem unreasonable considering that, without scatter corrections, errors in the reconstructed attenuation coefficients were between 18 to 45% depending on the phantom size and transmission source used.

La tomographie par émission de positrons est une technique d’imagerie nucléaire utilisant des noyaux radioactifs. Elle est utilisée dans le domaine clinique et préclinique. Cette dernière nécessite l’utilisation de petits animaux, comme la souris. Comme en imagerie clinique, l’objectif est d’obtenir le meilleur signal avec la meilleure précision spatiale possible. Cependant, un rapport d’échelle homme-souris suggère une résolution inférieure à 1 mm3. Un imageur conventionnel est constitué de modules de détection entourant le patient, orientés radialement. Cette approche lie efficacité et résolution spatiale. Ce travail concerne l’étude de la géométrie axiale. Les éléments de détection sont ici orientés parallèlement à l’objet. Ceci limite la corrélation entre efficacité de détection et résolution spatiale, et ainsi permet d’obtenir une haute résolution et haute sensibilité. La simulation de prototypes a permis d’envisager une résolution spatiale moyenne inférieure au millimètre et une efficacité de 15 ou 40% selon l’extension axiale. Ces résultats permettent de présager de bonnes perspectives en imageries préclinique et clinique. / Positrons emission tomography is a nuclear imaging technics using nuclear decays. It is used both in clinical and preclinical studies. The later requires the use of small animals such as the mouse. The objective is to obtain the best signal with the best spatial resolution. Yet, a weight ratio between humans and mice indicates the need of a sub-millimeter resolution. A conventional scanner is based on detection modules surrounding the object to image and arranged perpendicularly. This implies a strong relationship between efficiency and spatial resolution. This work focuses on the axial geometry in which detection modules are arranged parallel to the object. This limits the relationship between the figures of merit, leading to both high spatial resolution and efficiency. The simulations of prototypes showed great perspectives in term of sub-millimeter resolution with efficiencies of 15 or 40% according to the scanner’s axial extension. These results indicate great perspectives for both clinical and preclinical imaging.

The images in positron emission tomography (PET) represent three dimensional dynamic distributions of biologically interesting molecules labelled with positron emitting radionuclides (radiotracers). Spatial localisation of the radio-tracers is achieved by detecting in coincidence two collinear photons which are emitted when the positron annihilates with an ordinary electron. In order to obtain quantitatively accurate images in PET, it is necessary to correct for the effects of photon attenuation within the subject being imaged. These corrections can be obtained using singles-mode photon transmission scanning. Although suitable for small animal PET, these scans are subject to high amounts of contamination from scattered photons. Currently, no accurate correction exists to account for scatter in these data. The primary purpose of this work was to implement and validate an analytical scatter correction for PET transmission scanning. In order to isolate the effects of scatter, we developed a simulation tool which was validated using experimental transmission data. We then presented an analytical scatter correction for singles-mode transmission data in PET. We compared our scatter correction data with the previously validated simulation data for uniform and non-uniform phantoms and for two different transmission source radionuclides. Our scatter calculation correctly predicted the contribution from scattered photons to the simulated data for all phantoms and both transmission sources. We then applied our scatter correction as part of an iterative reconstruction algorithm for simulated and experimental PET transmission data for uniform and non-uniform phantoms. We also tested our reconstruction and scatter correction procedure using transmission data for several animal studies (mice, rats and primates). For all studies considered, we found that the average reconstructed linear attenuation coefficients for water or soft-tissue regions of interest agreed with expected values to within 4%. Using a 2.2 GHz processor, the scatter correction required between 6 to 27 minutes of CPU time (without any code optimisation) depending on the phantom size and source used. This extra calculation time does not seem unreasonable considering that, without scatter corrections, errors in the reconstructed attenuation coefficients were between 18 to 45% depending on the phantom size and transmission source used. / Science, Faculty of / Physics and Astronomy, Department of / Graduate

L’imagerie cardiovasculaire du rongeur par RMN est un véritable défi en ce qui concerne la résolution spatiale et temporelle, le contraste et le temps d’expérience. S’il est aujourd’hui admis que l’acquisition 3D doit être privilégiée chez le petit animal, les temps d’acquisition en 3D sont parfois très longs. Ils doivent pourtant rester compatibles avec les temps d’expérience in vivo. L’objectif de cette thèse était donc de développer de nouvelles méthodes d’imagerie cardiovasculaire 3D rapides pour le petit animal à 4.7 et 9.4 T. Tout d’abord, nous avons développé deux méthodes d’IRM cardiaque 4D (3D résolue dans le temps) à contraste «sang noir». La première méthode est basée sur une séquence TrueFISP (Fast Imaging with Steady-state Precession). Elle a permis d’obtenir le contraste sang noir en une heure d’acquisition. La deuxième méthode est basée sur une séquence FLASH (Fast Low Angle Shot). Elle utilise un gradient bipolaire pour supprimer le signal sanguin et le contraste a été rehaussé en Manganèse. Trente minutes d’acquisition ont alors été suffisantes. Ensuite, une méthode d’angiographie temps-de-vol 3D du corps entier de la souris a été développée. Le contraste vasculaire a été amélioré grâce à l’adjonction de motifs de suppression du signal tissulaire. L’imagerie de l’arbre vasculaire entier a pu être réalisé en moins de 10 minutes. Enfin, une nouvelle méthode d’angiographie fonctionnelle ciné temps-de-vol 4D utilisant une acquisition écho-planar a été développée. Les résultats préliminaires montrent qu’il est possible de diviser par quatre les temps d’acquisition de l’angiographie fonctionnelle classique. Tous ces résultats montrent que l’imagerie cardiovasculaire 3D haute résolution est possible dans des temps d’acquisition raisonnables voire rapides / Cardiovascular MRI in rodents is a real challenge in terms of spatial and temporal resolution, contrast and experiment times. Though it is accepted that 3D acquisition should be preferred in small animals, 3D acquisition times can be very long. However, they must remain compatible with in vivo experiment times. The aim of this thesis was therefore to develop new fast 3D methods of cardiovascular imaging in small animals at 4.7 and 9.4 T. First, two 4D cardiac MRI methods (3D time resolved) were developed in «black-blood» contrast. The first method is based on a TrueFISP sequence (Fast Imaging with Steady-state Precession). It allowed to make black blood contrast in one hour acquisition time. The second method is based on a FLASH sequence (Fast Low Angle Shot). It uses a bipolar gradient to suppress the blood signal and the contrast was enhanced by using Manganese. Thirty minutes were then enough. Next, a time-of-flight angiography method for the whole body of mice was developed. The vascular contrast was improved by adding preparation modules to suppress the signal from tissues. The imaging of the whole arterial tree was realized within less than ten minutes. Finally, a new 4D time-of-flight method of functional cine angiography with echo-planar acquisition was developed. Preliminary results showed that acquisition times could be divided by four compared with those in classical functional angiography. All these results show that high resolution 3D cardiovascular imaging is possible in reasonable or even fast acquisition times.

INTRODUCTION: SARS-CoV-2 is a novel virus from the coronavirus family that emerged in the Hubei province of China in December 2019 and rapidly spread throughout the world. On March 11, 2020, the World Health Organization declared a global pandemic. Infection with SARS-CoV-2 causes coronavirus disease 19 (COVID19) which can be fatal. There is an obvious and pressing need for research surrounding SARS-CoV-2 that will aid in eradication of this pandemic. OBJECTIVE: The goal of this study was to absolve the dire need for small animal models of human disease that demonstrate hallmark pathological features of infection. Due to ethical and financial obstacles, the use of animals that closely resemble human immunity, such as non-human primates, is often not a viable option. For this reason, there is a push to develop small animal models that can mimic human disease responses, particularly those in viral infections that have a narrow species tropism. To achieve this in the context of the novel coronavirus, SARS-CoV-2, we studied various mouse models and their capacity to become infected with and mount an immune response to SARS-CoV-2. Our goal was to identify a model that sufficiently mimics severe COVID19 seen in humans as well as provide molecular insight into pathways that prevent the development of severe disease. METHODS: NRG-L and HIS-NRGF-L mice were subcutaneously implanted with human fetal lung xenografts and infected with SARS-CoV-2. Tissues were stained with H&E for histopathological scoring. NRG-L and HIS-NRGF-L tissues were fluorescently labeled using 2 different multiplex immunohistochemistry panels. Slides were digitized by a Vectra Polaris™ fluorescent whole slide scanner and digital analysis was completed using HALO™. Statistical analysis was conducted using GraphPad Prism™ 9.0.1. RESULTS: Infected NRG-L mice present extensive histopathological manifestations when compared to uninfected controls. Cumulative histology scores at both 2 and 7DPI were increased when compared to uninoculated fLX. Neutrophil influx, intra-airspace necrosis, capillary fibrin thrombi, and presence of syncytial cells were the most significant independent observations that contributed to the increased cumulative score. Together these findings indicate that fLX inoculated with SARS-CoV-2 faithfully recapitulate several features of diffuse alveolar damage (DAD) described in severe COVID-19. HIS-NRGF-L mice displayed decreased influx of neutrophils, intra-airway necrosis, and syncytial cells when compared to NRG-L fLX. Hemorrhage was decreased at both 2 and 7 DPI for HIS-NRGF-L fLX compared to NRG-L fLX. Cumulative histology scores were decreased in HIS-NRGF-L fLX at 7 DPI when compared to NRG-L fLX. Taken together these findings support the hypothesis that human myeloid and lymphoid infiltrates suppress or prevent the disparate host response observed in NRGL-L fLX that manifested in pronounced diffuse alveolar damage. CONCLUSION: Using digital image analysis of multiplex immunohistochemistry paired with semi-quantitative histopathological scoring, this study characterized important hallmark lesions observed in severe COVID19 as seen in small animal models. NRG-L and HIS-NRGF-L mice that are subcutaneously implanted with human fetal lung xenografts are susceptible to infection with SARS-CoV-2 and can produce severe and moderate disease phenotypes respectively. Co-engraftment with human fetal lung tissue and human immune system components in HIS-NRGF-L mice suppresses the divergent host response that is observed in NRG-L mice. For this reason, NRG-L mice engrafted with fLX are an ideal small animal model of severe COVID19, whereas HIS-NRGF-L mice severe as a valuable and informative model for deciphering molecular mechanisms driving severe COVID-19 that will serve as targets for therapeutic development.