Expansion and Light-Sheet Microscopy for Nanoscale 3D Imaging.

Expansion Microscopy (ExM) and Light-Sheet Fluorescence Microscopy (LSFM) are forefront imaging techniques that enable high-resolution visualization of biological specimens. ExM enhances nanoscale investigation using conventional fluorescence microscopes, while LSFM offers rapid, minimally invasive imaging over large volumes. This review explores the joint advancements of ExM and LSFM, focusing on the excellent performance of the integrated modality obtained from the combination of the two, which is refer to as ExLSFM. In doing so, the chemical processes required for ExM, the tailored optical setup of LSFM for examining expanded samples, and the adjustments in sample preparation for accurate data collection are emphasized. It is delve into various specimen types studied using this integrated method and assess its potential for future applications. The goal of this literature review is to enrich the comprehension of ExM and LSFM, encouraging their wider use and ongoing development, looking forward to the upcoming challenges, and anticipating innovations in these imaging techniques.

Synthetic CT imaging for PET monitoring in proton therapy: a simulation study.

Objective.This study addresses a fundamental limitation of in-beam positron emission tomography (IB-PET) in proton therapy: the lack of direct anatomical representation in the images it produces. We aim to overcome this shortcoming by pioneering the application of deep learning techniques to create synthetic control CT images (sCT) from combining IB-PET and planning CT scan data.Approach.We conducted simulations involving six patients who underwent irradiation with proton beams. Leveraging the architecture of a visual transformer (ViT) neural network, we developed a model to generate sCT images of these patients using the planning CT scans and the inter-fractional simulated PET activity maps during irradiation. To evaluate the model's performance, a comparison was conducted between the sCT images produced by the ViT model and the authentic control CT images-serving as the benchmark.Main results.The structural similarity index was computed at a mean value across all patients of 0.91, while the mean absolute error measured 22 Hounsfield Units (HU). Root mean squared error and peak signal-to-noise ratio values were 56 HU and 30 dB, respectively. The Dice similarity coefficient exhibited a value of 0.98. These values are comparable to or exceed those found in the literature. More than 70% of the synthetic morphological changes were found to be geometrically compatible with the ones reported in the real control CT scan.Significance.Our study presents an innovative approach to surface the hidden anatomical information of IB-PET in proton therapy. Our ViT-based model successfully generates sCT images from inter-fractional PET data and planning CT scans. Our model's performance stands on par with existing models relying on input from cone beam CT or magnetic resonance imaging, which contain more anatomical information than activity maps.

Walk-through flat panel total-body PET: a patient-centered design for high throughput imaging at lower cost using DOI-capable high-resolution monolithic detectors.

PURPOSE: Long axial field-of-view (LAFOV) systems have a much higher sensitivity than standard axial field-of-view (SAFOV) PET systems for imaging the torso or full body, which allows faster and/or lower dose imaging. Despite its very high sensitivity, current total-body PET (TB-PET) throughput is limited by patient handling (positioning on the bed) and often a shortage of available personnel. This factor, combined with high system costs, makes it hard to justify the implementation of these systems for many academic and nearly all routine nuclear medicine departments. We, therefore, propose a novel, cost-effective, dual flat panel TB-PET system for patients in upright standing positions to avoid the time-consuming positioning on a PET-CT table; the walk-through (WT) TB-PET. We describe a patient-centered, flat panel PET design that offers very efficient patient throughput and uses monolithic detectors (with BGO or LYSO) with depth-of-interaction (DOI) capabilities and high intrinsic spatial resolution. We compare system sensitivity, component costs, and patient throughput of the proposed WT-TB-PET to a SAFOV (= 26 cm) and a LAFOV (= 106 cm) LSO PET systems. METHODS: Patient width, height (= top head to start of thighs) and depth (= distance from the bed to front of patient) were derived from 40 randomly selected PET-CT scans to define the design dimensions of the WT-TB-PET. We compare this new PET system to the commercially available Siemens Biograph Vision 600 (SAFOV) and Siemens Quadra (LAFOV) PET-CT in terms of component costs, system sensitivity, and patient throughput. System cost comparison was based on estimating the cost of the two main components in the PET system (Silicon Photomultipliers (SiPMs) and scintillators). Sensitivity values were determined using Gate Monte Carlo simulations. Patient throughput times (including CT and scout scan, patient positioning on bed and transfer) were recorded for 1 day on a Siemens Vision 600 PET. These timing values were then used to estimate the expected patient throughput (assuming an equal patient radiotracer injected activity to patients and considering differences in system sensitivity and time-of-flight information) for WT-TB-PET, SAFOV and LAFOV PET. RESULTS: The WT-TB-PET is composed of two flat panels; each is 70 cm wide and 106 cm high, with a 50-cm gap between both panels. These design dimensions were justified by the patient sizes measured from the 40 random PET-CT scans. Each panel consists of 14 × 20 monolithic BGO detector blocks that are 50 × 50 × 16 mm in size and are coupled to a readout with 6 × 6 mm SiPMs arrays. For the WT-TB-PET, the detector surface is reduced by a factor of 1.9 and the scintillator volume by a factor of 2.2 compared to LAFOV PET systems, while demonstrating comparable sensitivity and much better uniform spatial resolution (< 2 mm in all directions over the FOV). The estimated component cost for the WT-TB-PET is 3.3 × lower than that of a 106 cm LAFOV system and only 20% higher than the PET component costs of a SAFOV. The estimated maximum number of patients scanned on a standard 8-h working day increases from 28 (for SAFOV) to 53-60 (for LAFOV in limited/full acceptance) to 87 (for the WT-TB-PET). By scanning faster (more patients), the amount of ordered activity per patient can be reduced drastically: the WT-TB-PET requires 66% less ordered activity per patient than a SAFOV. CONCLUSIONS: We propose a monolithic BGO or LYSO-based WT-TB-PET system with DOI measurements that departs from the classical patient positioning on a table and allows patients to stand upright between two flat panels. The WT-TB-PET system provides a solution to achieve a much lower cost TB-PET approaching the cost of a SAFOV system. High patient throughput is increased by fast patient positioning between two vertical flat panel detectors of high sensitivity. High spatial resolution (< 2 mm) uniform over the FOV is obtained by using DOI-capable monolithic scintillators.

In-vivo range verification analysis with in-beam PET data for patients treated with proton therapy at CNAO.

Morphological changes that may arise through a treatment course are probably one of the most significant sources of range uncertainty in proton therapy. Non-invasive in-vivo treatment monitoring is useful to increase treatment quality. The INSIDE in-beam Positron Emission Tomography (PET) scanner performs in-vivo range monitoring in proton and carbon therapy treatments at the National Center of Oncological Hadrontherapy (CNAO). It is currently in a clinical trial (ID: NCT03662373) and has acquired in-beam PET data during the treatment of various patients. In this work we analyze the in-beam PET (IB-PET) data of eight patients treated with proton therapy at CNAO. The goal of the analysis is twofold. First, we assess the level of experimental fluctuations in inter-fractional range differences (sensitivity) of the INSIDE PET system by studying patients without morphological changes. Second, we use the obtained results to see whether we can observe anomalously large range variations in patients where morphological changes have occurred. The sensitivity of the INSIDE IB-PET scanner was quantified as the standard deviation of the range difference distributions observed for six patients that did not show morphological changes. Inter-fractional range variations with respect to a reference distribution were estimated using the Most-Likely-Shift (MLS) method. To establish the efficacy of this method, we made a comparison with the Beam's Eye View (BEV) method. For patients showing no morphological changes in the control CT the average range variation standard deviation was found to be 2.5 mm with the MLS method and 2.3 mm with the BEV method. On the other hand, for patients where some small anatomical changes occurred, we found larger standard deviation values. In these patients we evaluated where anomalous range differences were found and compared them with the CT. We found that the identified regions were mostly in agreement with the morphological changes seen in the CT scan.

A neural network-based algorithm for simultaneous event positioning and timestamping in monolithic scintillators.

Objective. Monolithic scintillator crystals coupled to silicon photomultiplier (SiPM) arrays are promising detectors for PET applications, offering spatial resolution around 1 mm and depth-of-interaction information. However, their timing resolution has always been inferior to that of pixellated crystals, while the best results on spatial resolution have been obtained with algorithms that cannot operate in real-time in a PET detector. In this study, we explore the capabilities of monolithic crystals with respect to spatial and timing resolution, presenting new algorithms that overcome the mentioned problems.Approach.Our algorithms were tested first using a simulation framework, then on experimentally acquired data. We tested an event timestamping algorithm based on neural networks which was then integrated into a second neural network for simultaneous estimation of the event position and timestamp. Both algorithms are implemented in a low-cost field-programmable gate array that can be integrated in the detector and can process more than 1 million events per second in real-time.Results.Testing the neural network for the simultaneous estimation of the event position and timestamp on experimental data we obtain 0.78 2D FWHM on the (x,y) plane, 1.2 depth-of-interaction FWHM and 156 coincidence time resolution on a25mm×25mm×8mm×LYSO monolith read-out by 643mm×3mmHamamatsu SiPMs.Significance.Our results show that monolithic crystals combined with artificial intelligence can rival pixellated crystals performance for time-of-flight PET applications, while having better spatial resolution and DOI resolution. Thanks to the use of very light neural networks, event characterization can be done on-line directly in the detector, solving the issues of scalability and computational complexity that up to now were preventing the use of monolithic crystals in clinical PET scanners.

Monte Carlo Characterization of the Trimage Brain PET System.

The TRIMAGE project aims to develop a brain-dedicated PET/MR/EEG (Positron Emission Tomography/Magnetic Resonance/Electroencephalogram) system that is able to perform simultaneous PET, MR and EEG acquisitions. The PET component consists of a full ring with 18 sectors. Each sector includes three square detector modules based on dual sstaggered LYSO:Ce matrices read out by SiPMs. Using Monte Carlo simulations and following NEMA (National Electrical Manufacturers Association) guidelines, image quality procedures have been applied to evaluate the performance of the PET component of the system. The performance are reported in terms of spatial resolution, uniformity, recovery coefficient, spill over ratio, noise equivalent count rate (NECR) and scatter fraction. The results show that the TRIMAGE system is at the top of the current brain PET technologies.

Performance evaluation of the IRIS XL-220 PET/CT system, a new camera dedicated to non-human primates.

BACKGROUND: Non-human primates (NHP) are critical in biomedical research to better understand the pathophysiology of diseases and develop new therapies. Based on its translational and longitudinal abilities along with its non-invasiveness, PET/CT systems dedicated to non-human primates can play an important role for future discoveries in medical research. The aim of this study was to evaluate the performance of a new PET/CT system dedicated to NHP imaging, the IRIS XL-220 developed by Inviscan SAS. This was performed based on the National Electrical Manufacturers Association (NEMA) NU 4-2008 standard recommendations (NEMA) to characterize the spatial resolution, the scatter fraction, the sensitivity, the count rate, and the image quality of the system. Besides, the system was evaluated in real conditions with two NHP with 18F-FDG and (-)-[18F]FEOBV which targets the vesicular acetylcholine transporter, and one rat using 18F-FDG. RESULTS: The full width at half maximum obtained with the 3D OSEM algorithm ranged between 0.89 and 2.11 mm in the field of view. Maximum sensitivity in the 400-620 keV and 250-750 keV energy windows were 2.37% (22 cps/kBq) and 2.81% (25 cps/kBq), respectively. The maximum noise equivalent count rate (NEC) for a rat phantom was 82 kcps at 75 MBq and 88 kcps at 75 MBq for energy window of 250-750 and 400-620 keV, respectively. For the monkey phantom, the maximum NEC was 18 kcps at 126 MBq and 19 kcps at 126 MBq for energy window of 250-750 and 400-620 keV, respectively. The IRIS XL provided an excellent quality of images in non-human primates and rats using 18F-FDG. The images acquired using (-)-[18F]FEOBV were consistent with those previously reported in non-human primates. CONCLUSIONS: Taken together, these results showed that the IRIS XL-220 is a high-resolution system well suited for PET/CT imaging in non-human primates.

Localization of anatomical changes in patients during proton therapy with in-beam PET monitoring: A voxel-based morphometry approach exploiting Monte Carlo simulations.

PURPOSE: In-beam positron emission tomography (PET) is one of the modalities that can be used for in vivo noninvasive treatment monitoring in proton therapy. Although PET monitoring has been frequently applied for this purpose, there is still no straightforward method to translate the information obtained from the PET images into easy-to-interpret information for clinical personnel. The purpose of this work is to propose a statistical method for analyzing in-beam PET monitoring images that can be used to locate, quantify, and visualize regions with possible morphological changes occurring over the course of treatment. METHODS: We selected a patient treated for squamous cell carcinoma (SCC) with proton therapy, to perform multiple Monte Carlo (MC) simulations of the expected PET signal at the start of treatment, and to study how the PET signal may change along the treatment course due to morphological changes. We performed voxel-wise two-tailed statistical tests of the simulated PET images, resembling the voxel-based morphometry (VBM) method commonly used in neuroimaging data analysis, to locate regions with significant morphological changes and to quantify the change. RESULTS: The VBM resembling method has been successfully applied to the simulated in-beam PET images, despite the fact that such images suffer from image artifacts and limited statistics. Three dimensional probability maps were obtained, that allowed to identify interfractional morphological changes and to visualize them superimposed on the computed tomography (CT) scan. In particular, the characteristic color patterns resulting from the two-tailed statistical tests lend themselves to trigger alarms in case of morphological changes along the course of treatment. CONCLUSIONS: The statistical method presented in this work is a promising method to apply to PET monitoring data to reveal interfractional morphological changes in patients, occurring over the course of treatment. Based on simulated in-beam PET treatment monitoring images, we showed that with our method it was possible to correctly identify the regions that changed. Moreover we could quantify the changes, and visualize them superimposed on the CT scan. The proposed method can possibly help clinical personnel in the replanning procedure in adaptive proton therapy treatments.

A Monte Carlo detector response model for the IRIS PET preclinical scanner.

PET preclinical studies require high spatial resolution due to the limited size of the animal under investigation. To achieve this target, iterative image reconstruction algorithms are commonly preferred over the analytical methods because they offer the possibility of accurately modeling the whole imaging process. In this work, we propose an accurate factorized system matrix for the INVISCAN IRIS preclinical PET scanner to be used with an iterative algorithm. The model includes two components: the geometric component and the detector response of the system. The main innovative aspect of the work is the creation of the detector matrix using a Monte Carlo simulation, with a particular focus on the optimization of the simulation process to reduce the calculation time. The new system model is compared with the current IRIS model to evaluate the image quality, following the NEMA Standards NU 4-2008. The comparison showed an enhancement of the image quality, in terms of uniformity and recovery coefficients. This work confirms that the inclusion of the detector response into the system model leads to improved reconstruction results.

An automatic algorithm to exploit the symmetries of the system response matrix in positron emission tomography iterative reconstruction.

Positron emission tomography (PET) iterative 3D reconstruction is a very computational demanding task. One of the main issues of the iterative reconstruction concerns the management of the system response matrix (SRM). The SRM models the relationship between the projection and the voxel space and its memory footprint can easily exceed hundreds of GB. Moreover, in order to make the reconstruction fast enough not to hinder its practical application, the SRM must be stored in the random access memory of the workstation used for the reconstruction. This issue is normally solved by implementing efficient storage schemes and by reducing the number of redundant patterns in the SRM through symmetries. However, finding a sufficient number of symmetries is often non-trivial and is typically performed using dedicated solutions that cannot be exported to different detectors and geometries. In this paper, an automatic approach to reduce the memory footprint of a pre-computed SRM is described. The proposed approach was named symmetry search algorithm (SSA) and consists in an algorithm that searches for some of the redundant patterns present in the SRM, leading to its lossy compression. This approach was built to detect translations, reflections and coordinates swap in voxel space. Therefore, it is particularly well suited for those scanners where some of the rotational symmetries are broken, e.g. small animal scanner where the modules are arranged in a polygonal ring made of few elements, and dual head planar PET systems. In order to validate this approach, the SSA is applied to the SRM of a preclinical scanner (the IRIS PET/CT). The data acquired by the scanner were reconstructed with a dedicated maximum likelihood estimation maximization algorithm with both the uncompressed and the compressed SRMs. The results achieved show that the information lost due to the SSA compression is negligible. Compression factors up to 52 when using the SSA together with manually inserted symmetries and up to 204 when using the SSA alone, can be obtained for the IRIS SRM. These results come without significant differences in the values and in the main quality metrics of the reconstructed images, i.e. spatial resolution and noise. Although the compression factors depend on the system considered, the SSA is applicable to any SRM and therefore it can be considered a general tool to reduce the footprint of a pre-computed SRM.

Performance evaluation of the LightPath imaging system for intra-operative Cerenkov luminescence imaging.

The performances of an intra-operative optical imaging system for Cerenkov luminescence imaging of resected tumor specimens were evaluated with phantom studies. The spatial resolution, the linearity of the measured signal with the activity concentration and the minimum detectable activity concentration were considered. A high linearity was observed over a broad range of activity concentration (R2⩾0.99 down to ∼40 kBq/ml of 18F-FDG). For 18F-FDG activity distributions 2 mm deep in biological tissue, the measured detection limit was 8 kBq/ml and a spatial resolution of 2.5 mm was obtained. The detection limit of the imaging system is comparable with clinical activity concentrations in tumor specimens, and the spatial resolution is compatible with clinical requirements.

Online proton therapy monitoring: clinical test of a Silicon-photodetector-based in-beam PET.

Particle therapy exploits the energy deposition pattern of hadron beams. The narrow Bragg Peak at the end of range is a major advantage but range uncertainties can cause severe damage and require online verification to maximise the effectiveness in clinics. In-beam Positron Emission Tomography (PET) is a non-invasive, promising in-vivo technique, which consists in the measurement of the ß+ activity induced by beam-tissue interactions during treatment, and presents the highest correlation of the measured activity distribution with the deposited dose, since it is not much influenced by biological washout. Here we report the first clinical results obtained with a state-of-the-art in-beam PET scanner, with on-the-fly reconstruction of the activity distribution during irradiation. An automated time-resolved quantitative analysis was tested on a lacrimal gland carcinoma case, monitored during two consecutive treatment sessions. The 3D activity map was reconstructed every 10 s, with an average delay between beam delivery and image availability of about 6 s. The correlation coefficient of 3D activity maps for the two sessions (above 0.9 after 120 s) and the range agreement (within 1 mm) prove the suitability of in-beam PET for online range verification during treatment, a crucial step towards adaptive strategies in particle therapy.

TRIMAGE: A dedicated trimodality (PET/MR/EEG) imaging tool for schizophrenia.

Simultaneous PET/MR/EEG (Positron Emission Tomography - Magnetic Resonance - Electroencephalography), a new tool for the investigation of neuronal networks in the human brain, is presented here within the framework of the European Union Project TRIMAGE. The trimodal, cost-effective PET/MR/EEG imaging tool makes use of cutting edge technology both in PET and in MR fields. A novel type of magnet (1.5T, non-cryogenic) has been built together with a PET scanner that makes use of the most advanced photodetectors (i.e., SiPM matrices), scintillators matrices (LYSO) and digital electronics. The combined PET/MR/EEG system is dedicated to brain imaging and has an inner diameter of 260 mm and an axial Field-of-View of 160 mm. It enables the acquisition and assessment of molecular metabolic information with high spatial and temporal resolution in a given brain simultaneously. The dopaminergic system and the glutamatergic system in schizophrenic patients are investigated via PET, the same physiological/pathophysiological conditions with regard to functional connectivity, via fMRI, and its electrophysiological signature via EEG. In addition to basic neuroscience questions addressing neurovascular-metabolic coupling, this new methodology lays the foundation for individual physiological and pathological fingerprints for a wide research field addressing healthy aging, gender effects, plasticity and different psychiatric and neurological diseases. The preliminary performances of two components of the imaging tool (PET and MR) are discussed. Initial results of the search of possible candidates for suitable schizophrenia biomarkers are also presented as obtained with PET/MR systems available to the collaboration.

Cerenkov luminescence imaging: physics principles and potential applications in biomedical sciences.

Cerenkov luminescence imaging (CLI) is a novel imaging modality to study charged particles with optical methods by detecting the Cerenkov luminescence produced in tissue. This paper first describes the physical processes that govern the production and transport in tissue of Cerenkov luminescence. The detectors used for CLI and their most relevant specifications to optimize the acquisition of the Cerenkov signal are then presented, and CLI is compared with the other optical imaging modalities sharing the same data acquisition and processing methods. Finally, the scientific work related to CLI and the applications for which CLI has been proposed are reviewed. The paper ends with some considerations about further perspectives for this novel imaging modality.

INSIDE in-beam positron emission tomography system for particle range monitoring in hadrontherapy.

The quality assurance of particle therapy treatment is a fundamental issue that can be addressed by developing reliable monitoring techniques and indicators of the treatment plan correctness. Among the available imaging techniques, positron emission tomography (PET) has long been investigated and then clinically applied to proton and carbon beams. In 2013, the Innovative Solutions for Dosimetry in Hadrontherapy (INSIDE) collaboration proposed an innovative bimodal imaging concept that combines an in-beam PET scanner with a tracking system for charged particle imaging. This paper presents the general architecture of the INSIDE project but focuses on the in-beam PET scanner that has been designed to reconstruct the particles range with millimetric resolution within a fraction of the dose delivered in a treatment of head and neck tumors. The in-beam PET scanner has been recently installed at the Italian National Center of Oncologic Hadrontherapy (CNAO) in Pavia, Italy, and the commissioning phase has just started. The results of the first beam test with clinical proton beams on phantoms clearly show the capability of the in-beam PET to operate during the irradiation delivery and to reconstruct on-line the beam-induced activity map. The accuracy in the activity distal fall-off determination is millimetric for therapeutic doses.

PET iterative reconstruction incorporating an efficient positron range correction method.

Positron range is one of the main physical effects limiting the spatial resolution of positron emission tomography (PET) images. If positrons travel inside a magnetic field, for instance inside a nuclear magnetic resonance (MR) tomograph, the mean range will be smaller but still significant. In this investigation we examined a method to correct for the positron range effect in iterative image reconstruction by including tissue-specific kernels in the forward projection operation. The correction method was implemented within STIR library (Software for Tomographic Image Reconstruction). In order to obtain the positron annihilation distribution of various radioactive isotopes in water and lung tissue, simulations were performed with the Monte Carlo package GATE [Jan et al. 2004 [1]] simulating different magnetic field intensities (0 T, 3 T, 9.5 T and 11 T) along the axial scanner direction. The positron range kernels were obtained for (68)Ga in water and lung tissue for 0 T and 3 T magnetic field voxellizing the annihilation coordinates into a three-dimensional matrix. The proposed method was evaluated using simulations of material-variant and material-invariant positron range corrections for the HYPERImage preclinical PET-MR scanner. The use of the correction resulted in sharper active region boundary definition, albeit with noise enhancement, and in the recovery of the true activity mean value of the hot regions. Moreover, in the case where a magnetic field is present, the correction accounts for the non-isotropy of the positron range effect, resulting in the recovery of resolution along the axial plane.

Accurate molecular imaging of small animals taking into account animal models, handling, anaesthesia, quality control and imaging system performance.

Small-animal imaging has become an important technique for the development of new radiotracers, drugs and therapies. Many laboratories have now a combination of different small-animal imaging systems, which are being used by biologists, pharmacists, medical doctors and physicists. The aim of this paper is to give an overview of the important factors in the design of a small animal, nuclear medicine and imaging experiment. Different experts summarize one specific aspect important for a good design of a small-animal experiment.

Cherenkov luminescence measurements with digital silicon photomultipliers: a feasibility study.

BACKGROUND: A feasibility study was done to assess the capability of digital silicon photomultipliers to measure the Cherenkov luminescence emitted by a ß source. Cherenkov luminescence imaging (CLI) is possible with a charge coupled device (CCD) based technology, but a stand-alone technique for quantitative activity measurements based on Cherenkov luminescence has not yet been developed. Silicon photomultipliers (SiPMs) are photon counting devices with a fast impulse response and can potentially be used to quantify ß-emitting radiotracer distributions by CLI. METHODS: In this study, a Philips digital photon counting (PDPC) silicon photomultiplier detector was evaluated for measuring Cherenkov luminescence. The PDPC detector is a matrix of avalanche photodiodes, which were read one at a time in a dark count map (DCM) measurement mode (much like a CCD). This reduces the device active area but allows the information from a single avalanche photodiode to be preserved, which is not possible with analog SiPMs. An algorithm to reject the noisiest photodiodes and to correct the measured count rate for the dark current was developed. RESULTS: The results show that, in DCM mode and at (10-13) °C, the PDPC has a dynamic response to different levels of Cherenkov luminescence emitted by a ß source and transmitted through an opaque medium. This suggests the potential for this approach to provide quantitative activity measurements. Interestingly, the potential use of the PDPC in DCM mode for direct imaging of Cherenkov luminescence, as a opposed to a scalar measurement device, was also apparent. CONCLUSIONS: We showed that a PDPC tile in DCM mode is able to detect and image a ß source through its Cherenkov radiation emission. The detector's dynamic response to different levels of radiation suggests its potential quantitative capabilities, and the DCM mode allows imaging with a better spatial resolution than the conventional event-triggered mode. Finally, the same acquisition procedure and data processing could be employed also for other low light levels applications, such as bioluminescence.

Quantitative micro-CT based coronary artery profiling using interactive local thresholding and cylindrical coordinates.

BACKGROUND: Micro-CT is an established imaging technique for high-resolution non-destructive assessment of vascular samples, which is gaining growing interest for investigations of atherosclerotic arteries both in humans and in animal models. However, there is still a lack in the definition of micro-CT image metrics suitable for comprehensive evaluation and quantification of features of interest in the field of experimental atherosclerosis (ATS). OBJECTIVE: A novel approach to micro-CT image processing for profiling of coronary ATS is described, providing comprehensive visualization and quantification of contrast agent-free 3D high-resolution reconstruction of full-length artery walls. METHODS: Accelerated coronary ATS has been induced by high fat cholesterol-enriched diet in swine and left coronary artery (LCA) harvested en bloc for micro-CT scanning and histologic processing. A cylindrical coordinate system has been defined on the image space after curved multiplanar reformation of the coronary vessel for the comprehensive visualization of the main vessel features such as wall thickening and calcium content. A novel semi-automatic segmentation procedure based on 2D histograms has been implemented and the quantitative results validated by histology. RESULTS: The potentiality of attenuation-based micro-CT at low kV to reliably separate arterial wall layers from adjacent tissue as well as identify wall and plaque contours and major tissue components has been validated by histology. Morphometric indexes from histological data corresponding to several micro-CT slices have been derived (double observer evaluation at different coronary ATS stages) and highly significant correlations (R2 > 0.90) evidenced. Semi-automatic morphometry has been validated by double observer manual morphometry of micro-CT slices and highly significant correlations were found (R2 > 0.92). CONCLUSION: The micro-CT methodology described represents a handy and reliable tool for quantitative high resolution and contrast agent free full length coronary wall profiling, able to assist atherosclerotic vessels morphometry in a preclinical experimental model of coronary ATS and providing a link between in vivo imaging and histology.