Quarter-millimeter-resolution molecular mouse imaging with U-SPECT⁺.

Limited spatial resolution of preclinical positron emission tomography (PET) and single-photon emission computed tomography (SPECT) has slowed down applications of molecular imaging in small animals. Here we present the latest-generation U-SPECT system (U-SPECT⁺, MILabs, Utrecht, the Netherlands) enabling radionuclide imaging of mice with quarter-millimeter resolution. The system was equipped with the newest high-resolution collimator with 0.25 mm diameter circular pinholes. It was calibrated with technetium-99 m point source measurements from which the system matrix was calculated. Images were reconstructed using pixel-based ordered subset expectation maximization (OSEM). Various phantoms and mouse SPECT scans were acquired. The reconstructed spatial resolution (the smallest visible capillary diameter in a hot-rod resolution phantom) was 0.25 mm. Knee joint images show tiny structures such as the femur epicondyle sulcus, as well as a clear separation between cortical and trabecular bone structures. In addition, time-activity curves of the lumbar spine illustrated that tracer dynamics in tiny tissue amounts could be measured. U-SPECT⁺ allows discrimination between molecular concentrations in adjacent volumes of as small as 0.015 µL, which is significantly better than can be imaged by any existing SPECT or PET system. This increase in the level of detail makes it more and more attractive to replace ex vivo methods and allows monitoring biological processes in tiny parts of organs in vivo.

Targeted multi-pinhole SPECT.

PURPOSE: Small-animal single photon emission computed tomography (SPECT) with focused multi-pinhole collimation geometries allows scanning modes in which large amounts of photons can be collected from specific volumes of interest. Here we present new tools that improve targeted imaging of specific organs and tumours, and validate the effects of improved targeting of the pinhole focus. METHODS: A SPECT system with 75 pinholes and stationary detectors was used (U-SPECT-II). An XYZ stage automatically translates the animal bed with a specific sequence in order to scan a selected volume of interest. Prior to stepping the animal through the collimator, integrated webcams acquire images of the animal. Using sliders, the user designates the desired volume to be scanned (e.g. a xenograft or specific organ) on these optical images. Optionally projections of an atlas are overlaid semiautomatically to locate specific organs. In order to assess the effects of more targeted imaging, scans of a resolution phantom and a mouse myocardial phantom, as well as in vivo mouse cardiac and tumour scans, were acquired with increased levels of targeting. Differences were evaluated in terms of count yield, hot rod visibility and contrast-to-noise ratio. RESULTS: By restricting focused SPECT scans to a 1.13-ml resolution phantom, count yield was increased by a factor 3.6, and visibility of small structures was significantly enhanced. At equal noise levels, the small-lesion contrast measured in the myocardial phantom was increased by 42%. Noise in in vivo images of a tumour and the mouse heart was significantly reduced. CONCLUSION: Targeted pinhole SPECT improves images and can be used to shorten scan times. Scan planning with optical cameras provides an effective tool to exploit this principle without the necessity for additional X-ray CT imaging.

Absolute quantitative total-body small-animal SPECT with focusing pinholes.

PURPOSE: In pinhole SPECT, attenuation of the photon flux on trajectories between source and pinholes affects quantitative accuracy of reconstructed images. Previously we introduced iterative methods that compensate for image degrading effects of detector and pinhole blurring, pinhole sensitivity and scatter for multi-pinhole SPECT. The aim of this paper is (1) to investigate the accuracy of the Chang algorithm in rodents and (2) to present a practical Chang-based method using body outline contours obtained with optical cameras. METHODS: Here we develop and experimentally validate a practical method for attenuation correction based on a Chang first-order method. This approach has the advantage that it is employed after, and therefore independently from, iterative reconstruction. Therefore, no new system matrix has to be calculated for each specific animal. Experiments with phantoms and animals were performed with a high-resolution focusing multi-pinhole SPECT system (U-SPECT-II, MILabs, The Netherlands). This SPECT system provides three additional optical camera images of the animal for each SPECT scan from which the animal contour can be estimated. RESULTS: Phantom experiments demonstrated that an average quantification error of -18.7% was reduced to -1.7% when both window-based scatter correction and Chang correction based on the body outline from optical images were applied. Without scatter and attenuation correction, quantification errors in a sacrificed rat containing sources with known activity ranged from -23.6 to -9.3%. These errors were reduced to values between -6.3 and +4.3% (with an average magnitude of 2.1%) after applying scatter and Chang attenuation correction. CONCLUSION: We conclude that the modified Chang correction based on body contour combined with window-based scatter correction is a practical method for obtaining small-animal SPECT images with high quantitative accuracy.

U-SPECT-II: An Ultra-High-Resolution Device for Molecular Small-Animal Imaging.

UNLABELLED: We present a new rodent SPECT system (U-SPECT-II) that enables molecular imaging of murine organs down to resolutions of less than half a millimeter and high-resolution total-body imaging. METHODS: The U-SPECT-II is based on a triangular stationary detector set-up, an XYZ stage that moves the animal during scanning, and interchangeable cylindric collimators (each containing 75 pinhole apertures) for both mouse and rat imaging. A novel graphical user interface incorporating preselection of the field of view with the aid of optical images of the animal focuses the pinholes to the area of interest, thereby maximizing sensitivity for the task at hand. Images are obtained from list-mode data using statistical reconstruction that takes system blurring into account to increase resolution. RESULTS: For (99m)Tc, resolutions determined with capillary phantoms were smaller than 0.35 and 0.45 mm using the mouse collimator with 0.35- and 0.6-mm pinholes, respectively, and less than 0.8 mm using the rat collimator with 1.0-mm pinholes. Peak geometric sensitivity is 0.07% and 0.18% for the mouse collimator with 0.35- and 0.6-mm pinholes, respectively, and 0.09% for the rat collimator. Resolution with (111)In, compared with that with (99m)Tc, was barely degraded, and resolution with (125)I was degraded by about 10%, with some additional distortion. In vivo, kidney, tumor, and bone images illustrated that U-SPECT-II could be used for novel applications in the study of dynamic biologic systems and radiopharmaceuticals at the suborgan level. CONCLUSION: Images and movies obtained with U-SPECT-II provide high-resolution radiomolecule visualization in rodents. Discrimination of molecule concentrations between adjacent volumes of about 0.04 microL in mice and 0.5 microL in rats with U-SPECT-II is readily possible.

High-resolution clustered pinhole (131)Iodine SPECT imaging in mice.

INTRODUCTION: High-resolution pre-clinical (131)I SPECT can facilitate development of new radioiodine therapies for cancer. To this end, it is important to limit resolution-degrading effects of pinhole edge penetration by the high-energy γ-photons of iodine. Here we introduce, optimize and validate (131)I SPECT performed with a dedicated high-energy clustered multi-pinhole collimator. METHODS: A SPECT-CT system (VECTor/CT) with stationary gamma-detectors was equipped with a tungsten collimator with clustered pinholes. Images were reconstructed with pixel-based OSEM, using a dedicated (131)I system matrix that models the distance- and energy-dependent resolution and sensitivity of each pinhole, as well as the intrinsic detector blurring and variable depth of interaction in the detector. The system performance was characterized with phantoms and in vivo static and dynamic (131)I-NaI scans of mice. RESULTS: Reconstructed image resolution reached 0.6mm, while quantitative accuracy measured with a (131)I filled syringe reaches an accuracy of +3.6±3.5% of the gold standard value. In vivo mice scans illustrated a clear shape of the thyroid and biodistribution of (131)I within the animal. Pharmacokinetics of (131)I was assessed with 15-s time frames from the sequence of dynamic images and time-activity curves of (131)I-NaI. CONCLUSIONS: High-resolution quantitative and fast dynamic (131)I SPECT in mice is possible by means of a high-energy collimator and optimized system modeling. This enables analysis of (131)I uptake even within small organs in mice, which can be highly valuable for development and optimization of targeted cancer therapies.

Optimizing modelling in iterative image reconstruction for preclinical pinhole PET.

The recently developed versatile emission computed tomography (VECTor) technology enables high-energy SPECT and simultaneous SPECT and PET of small animals at sub-mm resolutions. VECTor uses dedicated clustered pinhole collimators mounted in a scanner with three stationary large-area NaI(Tl) gamma detectors. Here, we develop and validate dedicated image reconstruction methods that compensate for image degradation by incorporating accurate models for the transport of high-energy annihilation gamma photons. Ray tracing software was used to calculate photon transport through the collimator structures and into the gamma detector. Input to this code are several geometric parameters estimated from system calibration with a scanning (99m)Tc point source. Effects on reconstructed images of (i) modelling variable depth-of-interaction (DOI) in the detector, (ii) incorporating photon paths that go through multiple pinholes ('multiple-pinhole paths' (MPP)), and (iii) including various amounts of point spread function (PSF) tail were evaluated. Imaging (18)F in resolution and uniformity phantoms showed that including large parts of PSFs is essential to obtain good contrast-noise characteristics and that DOI modelling is highly effective in removing deformations of small structures, together leading to 0.75 mm resolution PET images of a hot-rod Derenzo phantom. Moreover, MPP modelling reduced the level of background noise. These improvements were also clearly visible in mouse images. Performance of VECTor can thus be significantly improved by accurately modelling annihilation gamma photon transport.

U-SPECT-I: a novel system for submillimeter-resolution tomography with radiolabeled molecules in mice.

UNLABELLED: A major advance in biomedical science and diagnosis was accomplished with the development of in vivo techniques to image radiolabeled molecules, but limited spatial resolution has slowed down applications to small experimental animals. Here, we present a SPECT system (U-SPECT-I) dedicated to radionuclide imaging of murine organs at a submillimeter resolution. METHODS: The high performance of U-SPECT-I is based on a static triangular detector setup, with a cylindric imaging cavity in the center and 75 gold micropinhole apertures in the cavity wall. The pinholes are focused on a small volume of interest such as the mouse heart or spine to maximize the detection yield of gamma-photons. Three-dimensional molecular distributions are iteratively estimated using the detector data and a statistical reconstruction algorithm that takes into account system blurring and data noise to increase resolution and reduce image noise. RESULTS: With 0.6-mm-diameter pinholes, the maximum fraction of detected photons emitted by a point source (peak sensitivity) is 0.22% for a 15%-wide energy window and remains higher than 0.12% in the central 12 mm of the central plane. In a resolution phantom, radioactively filled capillaries as small as 0.5 mm and separated by 0.5 mm can be distinguished clearly in reconstructions. Projection data needed for the reconstruction of cross sections of molecular distributions in mouse organs can readily be obtained without the need for any mechanical movements. Images of a mouse spine show 99mTc-hydroxymethylene diphosphonate uptake down to the level of tiny parts of vertebral processes. These are separated clearly from the vertebral and intervertebral foramina. Using another tracer, one can monitor myocardial perfusion in the left and right ventricular walls, even in structures as small as the papillary muscles. CONCLUSION: U-SPECT-I allows discrimination between molecular concentrations in adjacent volumes of as small as about 0.1 muL, which is significantly smaller than can be imaged by any existing SPECT or PET system. Our initial in vivo images of the mouse heart and spine show that U-SPECT-I can be used for novel applications in the study of dynamic biologic systems with a clear projection to clinical applications. The combination of high resolution and detection efficiency of U-SPECT-I opens up new possibilities for the suborgan-level study of radiotracers in mouse models.

Ultra-high-sensitivity submillimeter mouse SPECT.

UNLABELLED: SPECT with submegabecquerel amounts of tracer or subsecond time resolution would enable a wide range of new imaging protocols such as screening tracers with initially low yield or labeling efficiency, imaging low receptor densities, or even performing SPECT outside regular radiation laboratories. To this end we developed dedicated ultra-high-sensitivity pinhole SPECT. METHODS: A cylindric collimator with 54 focused 2.0-mm-diameter conical pinholes was manufactured and mounted in a stationary small-animal SPECT system. The system matrix for image reconstruction was calculated via a hybrid method based on both (99m)Tc point source measurements and ray-tracing analytic modeling. SPECT images were reconstructed using pixel-based ordered-subsets expectation maximization. Performance was evaluated with phantoms and low-dose bone, dynamic kidney, and cardiac mouse scans. RESULTS: The peak sensitivity reached 1.3% (13,080 cps/MBq). The reconstructed spatial resolution (rod visibility in a micro-Jaszczak phantom) was 0.85 mm. Even with only a quarter megabecquerel of activity, 30-min bone SPECT scans provided surprisingly high levels of detail. Dynamic dual-isotope kidney and (99m)Tc-sestamibi cardiac scans were acquired with a time-frame resolution down to 1 s. CONCLUSION: The high sensitivity achieved increases the range of mouse SPECT applications by enabling in vivo imaging with less than a megabecquerel of tracer activity or down to 1-s frame dynamics.

Photon penetration and scatter in micro-pinhole imaging: a Monte Carlo investigation.

Pinhole SPECT is rapidly gaining popularity for imaging laboratory animals using gamma-emitting molecules. Penetration and scattering of gamma radiation in the pinhole edge material can account for a significant fraction of the total number of photons detected, particularly if the pinholes have small diameters. This study characterizes the effects of penetration and scatter with micro-pinholes made of lead, tungsten, gold and platinum. Monte Carlo simulations are performed for 1-125 (27-35 keV) and Tc-99m (140 keV) point sources with pinhole diameters ranging from 50 to 500 microm. The simulations account for the effects of photo-electric interaction, Rayleigh scattering, Compton scattering, ionization, bremsstrahlung and electron multiple scattering. As a typical example, in the case of a Tc-99m point source and pinholes with a diameter of 300 microm in gold or platinum, approximately 55% of the photons detected resulted from penetration and approximately 3% from scatter. For pinhole diameters ranging from 100 to 500 microm, the penetration fraction for tungsten and lead was approx a factor of 1.0 to 1.6 higher and the scatter fraction was 1.0 to 1.8 times higher than in case of gold or platinum. Using I-125 instead of Tc-99m decreases the penetration fraction by a factor ranging from 3 to 11 and the scatter fraction by a factor ranging from 12 to 40. For all materials studied, the total amounts of penetrated and scattered photons changed approximately linearly with respect to the pinhole diameter.

Performance assessment of a preclinical PET scanner with pinhole collimation by comparison to a coincidence-based small-animal PET scanner.

UNLABELLED: PET imaging of rodents is increasingly used in preclinical research, but its utility is limited by spatial resolution and signal-to-noise ratio of the images. A recently developed preclinical PET system uses a clustered-pinhole collimator, enabling high-resolution, simultaneous imaging of PET and SPECT tracers. Pinhole collimation strongly departs from traditional electronic collimation achieved via coincidence detection in PET. We investigated the potential of such a design by direct comparison to a traditional PET scanner. METHODS: Two small-animal PET scanners, 1 with electronic collimation and 1 with physical collimation using clustered pinholes, were used to acquire data from Jaszczak (hot rod) and uniform phantoms. Mouse brain imaging using (18)F-FDG PET was performed on each system and compared with quantitative ex vivo autoradiography as a gold standard. Bone imaging using (18)F-NaF allowed comparison of imaging in the mouse body. Images were visually and quantitatively compared using measures of contrast and noise. RESULTS: Pinhole PET resolved the smallest rods (diameter, 0.85 mm) in the Jaszczak phantom, whereas the coincidence system resolved 1.1-mm-diameter rods. Contrast-to-noise ratios were better for pinhole PET when imaging small rods (<1.1 mm) for a wide range of activity levels, but this reversed for larger rods. Image uniformity on the coincidence system (<3%) was superior to that on the pinhole system (5%). The high (18)F-FDG uptake in the striatum of the mouse brain was fully resolved using the pinhole system, with contrast to nearby regions equaling that from autoradiography; a lower contrast was found using the coincidence PET system. For short-duration images (low-count), the coincidence system was superior. CONCLUSION: In the cases for which small regions need to be resolved in scans with reasonably high activity or reasonably long scan times, a first-generation clustered-pinhole system can provide image quality in terms of resolution, contrast, and the contrast-to-noise ratio superior to a traditional PET system.

Three-dimensional histologic validation of high-resolution SPECT of antibody distributions within xenografts.

UNLABELLED: Longitudinal imaging of intratumoral distributions of antibodies in vivo in mouse cancer models is of great importance for developing cancer therapies. In this study, multipinhole SPECT with sub-half-millimeter resolution was tested for exploring intratumoral distributions of radiolabeled antibodies directed toward the epidermal growth factor receptor (EGFr) and compared with full 3-dimensional target expression assessed by immunohistochemistry. METHODS: (111)In-labeled zalutumumab, a human monoclonal human EGFr-targeting antibody, was administered at a nonsaturating dose to 3 mice with xenografted A431 tumors exhibiting high EGFr expression. Total-body and focused in vivo tumor SPECT was performed at 0 and 48 h after injection and compared both visually and quantitatively with full 3-dimensional immunohistochemical staining for EGFr target expression. RESULTS: SPECT at 48 h after injection showed that activity was predominantly concentrated in the tumor (10.5% ± 1.3% of the total-body activity; average concentration, 30.1% ± 4.6% of the injected dose per cubic centimeter). (111)In-labeled EGFr-targeting antibodies were distributed heterogeneously throughout the tumor. Some hot spots were observed near the tumor rim. Immunohistochemistry indicated that the antibody distributions obtained by SPECT were morphologically similar to those obtained for ex vivo EGFr target expression. Regions showing low SPECT activity were necrotic or virtually negative for EGFr target expression. A good correlation (r = 0.86, P < 0.0001) was found between the percentage of regions showing low activity on SPECT and the percentage of necrotic tissue on immunohistochemistry. CONCLUSION: Multipinhole SPECT enables high-resolution visualization and quantification of the heterogeneity of (111)In-zalutumumab concentrations in vivo.

VECTor: a preclinical imaging system for simultaneous submillimeter SPECT and PET.

UNLABELLED: Today, PET and SPECT tracers cannot be imaged simultaneously at high resolutions but require 2 separate imaging systems. This paper introduces a Versatile Emission Computed Tomography system (VECTor) for radionuclides that enables simultaneous submillimeter imaging of single-photon and positron-emitting radiolabeled molecules. METHODS: γ-photons produced both by electron-positron annihilation and by single-photon emitters are projected onto the same detectors by means of a novel cylindric high-energy collimator containing 162 narrow pinholes that are grouped in clusters. This collimator is placed in an existing SPECT system (U-SPECT-II) with 3 large-field-of-view γ-detectors. From the acquired projections, PET and SPECT images are obtained using statistical image reconstruction that corrects for energy-dependent system blurring. RESULTS: For PET tracers, the tomographic resolution obtained with a Jaszczak hot rod phantom was less than 0.8 mm, and 0.5-mm resolution images of SPECT tracers were acquired simultaneously. SPECT images were barely degraded by the simultaneous presence of a PET tracer, even when the activity concentration of the PET tracer exceeded that of the SPECT tracer by up to a factor of 2.5. Furthermore, we simultaneously acquired fully registered 3- and 4-dimensional multiple functional images from living mice that, in the past, could be obtained only sequentially. CONCLUSION: High-resolution complementary information about tissue function contained in SPECT and PET tracer distributions can now be obtained simultaneously using a fully integrated imaging device. These combined unique capabilities pave the way for new perspectives in imaging the biologic systems of rodents.

Murine cardiac images obtained with focusing pinhole SPECT are barely influenced by extra-cardiac activity.

Ultra-high-resolution SPECT images can be obtained with focused multipinhole collimators. Here we investigate the influence of unwanted high tracer uptake outside the scan volume on reconstructed tracer distributions inside the scan volume, for (99m)Tc-tetrofosmin myocardial perfusion scanning in mice. Simulated projections of a digital mouse phantom (MOBY) in a focusing multipinhole SPECT system (U-SPECT-II, MILabs, The Netherlands) were generated. With this system differently sized user-defined scan volumes can be selected, by translating the animal in 3D through the focusing collimators. Scan volume selections were set to (i) a minimal volume containing just the heart, acquired without translating the animal during scanning, (ii) a slightly larger scan volume as is typically applied for the heart, requiring only small XYZ translations during scanning, (iii) same as (ii), but extended further transaxially, and (iv) same as (ii), but extended transaxially to cover the full thorax width (gold standard). Despite an overall negative bias that is significant for the minimal scan volume, all selected volumes resulted in visually similar images. Quantitative differences in the reconstructed myocardium between gold standard and the results from the smaller scan volume selections were small; the 17 standardized myocardial segments of a bull's eye plot, normalized to the myocardial mean of the gold standard, deviated on average 6.0%, 2.5% and 1.9% for respectively the minimal, the typical and the extended scan volume, while maximum absolute deviations were respectively 18.6%, 9.0% and 5.2%. Averaged over ten low-count noisy simulations, the mean absolute deviations were respectively 7.9%, 3.2% and 1.9%. In low-count noisy simulations, the mean and maximum absolute deviations for the minimal scan volume could be reduced to respectively 4.2% and 12.5% by performing a short survey scan of the exterior activity and focusing the remaining scan time at the organ of interest. We conclude that reconstructed tracer distribution in the myocardium can be influenced by activity in surrounding organs when a too narrow scan volume is used. With slightly larger scan volumes this problem is adequately suppressed. This approach produced a smaller mean deviation and may be more effective than employing a narrow scan volume with an additional survey scan.

Fast spiral SPECT with stationary γ-cameras and focusing pinholes.

UNLABELLED: Small-animal SPECT systems with stationary detectors and focusing multiple pinholes can achieve excellent resolution-sensitivity trade-offs. These systems are able to perform fast total-body scans by shifting the animal bed through the collimator using an automated xyz stage. However, so far, a large number of highly overlapping central fields of view have been used, at the cost of overhead time needed for animal repositioning and long image reconstruction times due to high numbers of projection views. METHODS: To improve temporal resolution and reduce image reconstruction time for such scans, we have developed and tested spiral trajectories (STs) of the animal bed requiring fewer steps. In addition, we tested multiplane trajectories (MPTs) of the animal bed, which is the standard acquisition method of the U-SPECT-II system that is used in this study. Neither MPTs nor STs require rotation of the animal. Computer simulations and physical phantom experiments were performed for a wide range of numbers of bed positions. Furthermore, we tested STs in vivo for fast dynamic mouse scans. RESULTS: We found that STs require less than half the number of bed positions of MPTs to achieve sufficient sampling. The reduced number of bed positions made it possible to perform a dynamic total-body bone scan and a dynamic hepatobiliary scan with time resolutions of 60 s and 15 s, respectively. CONCLUSION: STs open up new possibilities for high throughput and fast dynamic radio-molecular imaging.

Varenicline increases in vivo striatal dopamine D2/3 receptor binding: an ultra-high-resolution pinhole [123I]IBZM SPECT study in rats.

INTRODUCTION: Ex vivo storage phosphor imaging rat studies reported increased brain dopamine D2/3 receptor (DRD2/3) availability following treatment with varenicline, a nicotinergic drug. However, ex vivo studies can only be performed using cross-sectional designs. Small-animal imaging offers the opportunity to perform serial assessments. We evaluated whether high-resolution pinhole single photon emission computed tomography (SPECT) imaging in rats was able to reproduce previous ex vivo findings. METHODS: Rats were imaged for baseline striatal DRD2/3 availability using ultra-high-resolution pinhole SPECT (U-SPECT-II) and [123I]IBZM as a radiotracer, and randomized to varenicline (n=7; 2 mg/kg) or saline (n=7). Following 2 weeks of treatment, a second scan was acquired. RESULTS: Significantly increased striatal DRD2/3 availability was found following varenicline treatment compared to saline (time⁎treatment effect): posttreatment difference in binding potential between groups corrected for initial baseline differences was 2.039 (P=.022), indicating a large effect size (d=1.48). CONCLUSIONS: Ultra-high-resolution pinhole SPECT can be used to assess varenicline-induced changes in DRD2/3 availability in small laboratory animals over time. Future small-animal studies should include imaging techniques to enable repeated within-subjects measurements and reduce the amount of animals.

Accurate Coregistration between Ultra-High-Resolution Micro-SPECT and Circular Cone-Beam Micro-CT Scanners.

Introduction. Spatially registering SPECT with CT makes it possible to anatomically localize SPECT tracers. In this study, an accurate method for the coregistration of ultra-high-resolution SPECT volumes and multiple cone-beam CT volumes is developed and validated, which does not require markers during animal scanning. Methods. Transferable animal beds were developed with an accurate mounting interface. Simple calibration phantoms make it possible to obtain both the spatial transformation matrix for stitching multiple CT scans of different parts of the animal and to register SPECT and CT. The spatial transformation for image coregistration is calculated once using Horn's matching algorithm. Animal images can then be coregistered without using markers. Results. For mouse-sized objects, average coregistration errors between SPECT and CT in X, Y, and Z directions are within 0.04 mm, 0.10 mm, and 0.19 mm, respectively. For rat-sized objects, these numbers are 0.22 mm, 0.14 mm, and 0.28 mm. Average 3D coregistration errors were within 0.24 mm and 0.42 mm for mouse and rat imaging, respectively. Conclusion. Extending the field-of-view of cone-beam CT by stitching is improved by prior registration of the CT volumes. The accuracy of registration between SPECT and CT is typically better than the image resolution of current ultra-high-resolution SPECT.

System calibration and statistical image reconstruction for ultra-high resolution stationary pinhole SPECT.

For multipinhole single-photon emission computed tomography (SPECT), iterative reconstruction algorithms are preferred over analytical methods, because of the often complex multipinhole geometries and the ability of iterative algorithms to compensate for effects like spatially variant sensitivity and resolution. Ideally, such compensation methods are based on accurate knowledge of the position-dependent point spread functions (PSFs) specifying the response of the detectors to a point source at every position in the instrument. This paper describes a method for model-based generation of complete PSF lookup tables from a limited number of point-source measurements for stationary SPECT systems and its application to a submillimeter resolution stationary small-animal SPECT system containing 75 pinholes (U-SPECT-I). The method is based on the generalization over the entire object to be reconstructed, of a small number of properties of point-source responses which are obtained at a limited number of measurement positions. The full shape of measured point-source responses can almost be preserved in newly created PSF tables. We show that these PSFs can be used to obtain high-resolution SPECT reconstructions: the reconstructed resolutions judged by rod visibility in a micro-Derenzo phantom are 0.45 mm with 0.6-mm pinholes and below 0.35 mm with 0.3-mm pinholes. In addition, we show that different approximations, such as truncating the PSF kernel, with significant reduction of reconstruction time, can still lead to acceptable reconstructions.

The pinhole: gateway to ultra-high-resolution three-dimensional radionuclide imaging.

Today the majority of clinical molecular imaging procedures are carried out with single-photon emitters and gamma cameras, in planar mode and single-photon emission computed tomography (SPECT) mode. Thanks to the development of advanced multi-pinhole collimation technologies, SPECT imaging of small experimental animals is rapidly gaining in popularity. Whereas resolutions in routine clinical SPECT are typically larger than 1 cm (corresponding to >1,000 microl), it has recently proved possible to obtain spatial resolutions of about 0.35 mm ( approximately 0.04 microl) in the mouse. Meanwhile, SPECT systems that promise an even better performance are under construction. The new systems are able to monitor functions in even smaller structures of the mouse than was possible with dedicated small animal positron emission tomography ( approximately 1 mm resolution, corresponding to 1 microl). This paper provides a brief history of image formation with pinholes and explains the principles of pinhole imaging and pinhole tomography and the basics of modern image reconstruction methods required for such systems. Some recently introduced ultra-high-resolution small animal SPECT instruments are discussed and new avenues for improving system performance are explored. This may lead to many completely new biomedical applications. We also demonstrate that clinical SPECT systems with focussing pinhole gamma cameras will be able to produce images with a resolution that may become superior to that of PET for major clinical applications. A design study of a cardiac pinhole SPECT system indicates that the heart can be imaged an order of magnitude faster or with much more detail than is possible with currently used parallel-hole SPECT (e.g. 3-4 mm instead of approximately 8 mm system resolution).