Michael J. Paulus

A new X-ray computed tomography system for laboratory mouse imaging

Two versions of a new high-resolution X-ray computed tomography system are being developed to screen mutagenized mice in the Oak Ridge National Laboratory Mammalian Genetics Research Facility. The first prototype employs a single-pixel CdZnTe detector with a pinhole collimator operating in pulse counting mode. The second version employs a phosphor screen/CCD detector operating in current mode. The major system hardware includes a low-energy X-ray tube, two linear translation stages and a rotational stage. For the single-pixel detector, image resolution is determined by the step size of the detector stage; preliminary images have been acquired at 100 /spl mu/m and 250 /spl mu/m resolutions. The resolution of the phosphor screen detector is determined by the modulation transfer function of the phosphor screen; images with resolutions approaching 50 /spl mu/m have been acquired. The system performance with the two detectors is described and recent images are presented.

High resolution computed tomography and MRI for monitoring lung tumor growth in mice undergoing radioimmunotherapy: Correlation with histology

A model lung tumor system has been developed in mice for the evaluation of vascular targeted radioimmunotherapy. In this model, EMT-6 mammary carcinoma tumors growing in the lung are treated with 213Bi, an alpha particle emitter, which is targeted to lung blood vessels using a monoclonal antibody. Smaller tumors (< 100 microm in diameter) are cured, but larger tumors undergo a period of regression and then regrow and ultimately prove lethal. The goal of this work was to determine if external imaging with MRI or CT could be used routinely to monitor the growth/ regression of lung tumors in live mice. To attempt to evaluate individual tumors in vivo, animals were initially imaged with magnetic resonance imaging (MRI). High resolution MRI images could be obtained only after sacrifice when lungs were not moving. In contrast, high resolution computed tomography (CT) produced evaluable images from anesthetized animals. Serial CT images (up to 5/animal) were collected over a 17 day period of tumor growth and treatment. When tumored animals became moribund, animals were sacrificed and lungs were inflated with fixative, embedded in paraffin, and then sectioned serially to compare the detection of tumors by high resolution CT with detection by histology. CT proved most useful in detecting lung tumors located in the hilar area and least useful in detecting serosal surface and anterior lobe tumor foci. Overall, CT images of live animals revealed tumors in approximately 2/3 of cases detected in histologic serial sections when relatively few tumors were present per lung. Detection of lesions and their resolution post therapy were complicated due to residual hemorrhagic, regressing tumor nodules and the development of lung edema both of which appeared as high density areas in the CT scans. We conclude that the microCT method used could identify some lung tumors as small as 100 microm in diameter; however, no concrete evaluation of therapy induced regression of the tumors could be made with CT analyses alone.

Transgenic Mouse Model of AA Amyloidosis

AA amyloidosis can be induced in mice experimentally through injection of certain chemical or biological compounds. However, the usefulness of this approach is limited by its dependence on exogenous inflammatory agents that stimulate cytokines to increase the synthesis of precursor serum amyloid A (SAA) protein and the transitory nature of the pathological fibrillar deposits. We now report that transgenic mice carrying the human interleukin 6 gene under the control of the metallothionein-I promoter had markedly increased concentrations of SAA and developed amyloid in the spleen, liver, and kidneys by 3 months of age. At the time of death about 6 months later, organs obtained from these animals had extensive amyloid deposits. This disease process was apparent radiographically using small-animal computer axial tomography and magnetic resonance imaging equipment. The AA nature of the amyloid was evidenced immunohistochemically and was unequivocally established by sequence analysis of protein extracted from the fibrils. The availability of this unique in vivo experimental model of AA amyloidosis provides the means to assess the therapeutic efficacy of agents designed to reduce or prevent the fibrillar deposits found in AA and other types of amyloid-associated disease.

A comparison of x-ray detectors for mouse CT imaging

There is significant interest in using computed tomography (CT) for in vivo imaging applications in mouse models of disease. Most commercially available mouse x-ray CT scanners utilize a charge-coupled device (CCD) detector coupled via fibre optic taper to a phosphor screen. However, there has been little research to determine if this is the optimum detector for the specific task of in vivo mouse imaging. To investigate this issue, we have evaluated four detectors, including an amorphous selenium (a-Se) detector, an amorphous silicon (a-Si) detector with a gadolinium oxysulphide (GOS) screen, a CCD with a 3:1 fibre taper and a GOS screen, and a CCD with a 2:1 fibre taper and both GOS and thallium-doped caesium iodide (CsI:Tl) screens. The detectors were evaluated by measuring the modulation transfer function (MTF), noise power spectrum (NPS), detective quantum efficiency (DQE), stability over multiple exposures, and noise in reconstructed CT images. The a-Se detector had the best MTF and the highest DQE (0.6 at 0 lp mm(-1)) but had the worst stability (45% reduction after 2000 exposure frames). The a-Si detector and the CCD with the 3:1 fibre, both of which used the GOS screen, had very similar performance with a DQE of approximately 0.30 at 0 lp mm(-1). For the CCD with the 2:1 fibre, the CsI:Tl screen resulted in a nearly two-fold improvement in DQE over the GOS screen (0.4 versus 0.24 at 0 lp mm(-1)). The CCDs both had the best stability, with less than a 1% change in pixel values over multiple exposures. The pixel values of the a-Si detector increased 5% over multiple exposures due to the effects of image lag. Despite the higher DQE of the a-Se detector, the reconstructed CT images acquired with the a-Si detector had lower noise levels, likely due to the blurring effects from the phosphor screen.

Reconstruction of multi-energy X-ray computed tomography images of laboratory mice

A new X-ray computed tomography (CT) system is being developed at Oak Ridge National Laboratory to image laboratory mice for the purpose of rapid phenotype screening and identification. One implementation of this CT system allows simultaneous capture of several sets of sinogram data, each having a unique X-ray energy distribution. The goals of this paper are to (1) identify issues associated with the reconstruction of this energy-dependent data and (2) suggest preliminary approaches to address these issues. Due to varying numbers of photon counts within each set, both traditional (filtered backprojection, or FBP) and statistical (maximum likelihood, or ML) tomographic image reconstruction techniques have been applied to the energy-dependent sinogram data. Results of reconstructed images using both algorithms on sinogram data (high- and low-count) are presented. Also, tissue contrast within the energy-dependent images is compared to known X-ray attenuation coefficients of soft tissue (e.g. muscle, bone, and fat).

A restraint-free small animal SPECT imaging system with motion tracking

We report on an approach toward the development of a high-resolution single photon emission computed tomography (SPECT) system to image the biodistribution of radiolabeled tracers such as Tc-99m and I-125 in unrestrained/unanesthetized mice. An infrared (IR)-based position tracking apparatus has been developed and integrated into a SPECT gantry. The tracking system is designed to measure the spatial position of a mouses head at a rate of 10-15 frames per second with submillimeter accuracy. The high-resolution, gamma imaging detectors are based on pixellated NaI(Tl) crystal scintillator arrays, position-sensitive photomultiplier tubes, and novel readout circuitry requiring fewer analog-digital converter (ADC) channels while retaining high spatial resolution. Two SPECT gamma camera detector heads based upon position-sensitive photomultiplier tubes have been built and installed onto the gantry. The IR landmark-based pose measurement and tracking system is under development to provide animal position data during a SPECT scan. The animal position and orientation data acquired by the tracking system will be used for motion correction during the tomographic image reconstruction.

Quantitative high-resolution microradiographic imaging of amyloid deposits in a novel murine model of AA amyloidosis

The mouse model of experimentally induced systemic AA amyloidosis is long established, well validated, and closely analogous to the human form of this disease. However, the induction of amyloid by experimental inflammation is unpredictable, inconsistent, and difficult to modulate. We have previously shown that murine AA amyloid deposits can be imaged using iodine-123 labeled SAP scintigraphy and report here substantial refinements in both the imaging technology and the mouse model itself. In this regard, we have generated a novel prototype of AA amyloid in which mice expressing the human interleukin 6 gene, when given amyloid enhancing factor, develop extensive and progressive systemic AA deposition without an inflammatory stimulus, i.e., a transgenic rapidly inducible amyloid disease (TRIAD) mouse. Additionally, we have constructed high-resolution micro single photon emission computed tomography (SPECT)/computed tomography (CT) instrumentation that provides images revealing the precise anatomic location of amyloid deposits labeled by radioiodinated serum amyloid P component (SAP). Based on reconstructed microSPECT/CT images, as well as autoradiographic, isotope biodistribution, and quantitative histochemical analyses, the 125I-labeled SAP tracer bound specifically to hepatic and splenic amyloid in the TRIAD animals. The ability to discern radiographically the extent of amyloid burden in the TRIAD model provides a unique opportunity to evaluate the therapeutic efficacy of pharmacologic compounds designed to inhibit fibril formation or effect amyloid resolution.

Micro-Imaging of Amyloid in Mice

Scintigraphic imaging of radioiodinated serum amyloid P-component is a proven method for the clinical detection of peripheral amyloid deposits (Hawkins et al., 1990). However, the inability to perform comparably high-resolution studies in experimental animal models of amyloid disease has impacted not only basic studies into the pathogenesis of amyloidosis but also in the preclinical in vivo evaluation of potential anti-amyloid therapeutic agents. We have developed microimaging technologies, implemented novel computational methods, and established protocols to generate high-resolution images of amyloid deposits in mice. (125)I-labeled serum amyloid P component (SAP) and an amyloid-fibril reactive murine monoclonal antibody (designated 11-1F4) have been used successfully to acquire high-resolution single photon emission computed tomographic (SPECT) images that, when fused with x-ray computed tomographic (CT) data, have provided precise anatomical localization of secondary (AA) and primary (AL) amyloid deposits in mouse models of these diseases. This chapter will provide detailed protocols for the radioiodination and purification of amyloidophilic proteins and the generation of mouse models of AA and AL amyloidosis. A brief description of the available hardware and the parameters used to acquire high-resolution microSPECT and CT images is presented, and the tools used to perform image reconstruction and visualization that permit the analysis and presentation of image data are discussed. Finally, we provide established methods for measuring organ- and tissue-specific activities with which to corroborate the microSPECT and CT images.

Fast Feldkamp reconstruction based on focus of attention and distributed computing

The Feldkamp algorithm is widely accepted as a practical conebeam reconstruction method for three‐dimensional x‐ray computed tomography. We introduce focus of attention, an effective and simple to implement datadriven preprocessing scheme, for identifying a convex subset of voxels that include all those relevant to the object under study. By concentrating on this subset of voxels during reconstruction, we reduce the computational demands of the Feldkamp algorithm correspondingly. To achieve further speed‐up, all computations are distributed across a cluster of inexpensive, dual‐processor PCs. We present experimental work based on mouse data obtained from the MicroCAT which is a high‐resolution x‐ray computed tomography system for small animal imaging. This work shows that focus of attention can cut the overall computation time in half without affecting the image quality. The method is general by nature and can easily be adapted to apply to other geometries and modalities as well as to iterative reconstruction algorithms.

A low-noise, wideband, integrated CMOS transimpedance preamplifier for photodiode applications

A low-noise, wideband, integrated CMOS transimpedance preamplifier is presented for silicon avalanche photodiode (APD) applications. The preamplifier, fabricated in a standard 2- mu CMOS technology, features a transimpedance gain of 45 k Omega , a risetime of 22 ns, a series noise of 1.6 nV/Hz/sup 1/2/, and a wideband equivalent input-noise current of 12 nA for a source capacitance of 12 pF. The measured /sup 22/Na timing resolution of 9.2-ns full width at half maximum (FWHM) and energy resolution of 22.4% FWHM for the RCA C30994 BGO/APD detector module coupled to the preamplifier are comparable to the performance reported using charge-sensitive preamplifiers. This shows that transimpedance preamplifiers should be considered for APD applications, especially where APD noise current dominates noise from feedback resistors in the 10-k Omega to 50-k Omega range. The transimpedance preamplifier reported here offers advantages of being fully monolithically integrated, having low power dissipation (38 mW), having low bandwidth sensitivity to source capacitance, requiring no shaping-amplifier pole-zero compensation, and requiring no feedback capacitance reset at high count rates.<<ETX>>