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 mL in mice and 0.5 mL in rats with U-SPECT-II is readily possible.

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).

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: g-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 g-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.

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.