Liji Cao

Bayesian reconstruction strategy of fluorescence-mediated tomography using an integrated SPECT-CT-OT system

Following the assembly of a triple-modality SPECT-CT-OT small animal imaging system providing intrinsically co-registered projection data of all three submodalities and under the assumption and investigation of dual-labeled probes consisting of both fluorophores and radionuclides, a novel multi-modal reconstruction strategy is presented in this paper aimed at improving fluorescence-mediated tomography (FMT). The following reconstruction procedure is proposed: firstly, standard x-ray CT image reconstruction is performed employing the FDK algorithm. Secondly, standard SPECT image reconstruction is performed using OSEM. Thirdly, from the reconstructed CT volume data the surface boundary of the imaged object is extracted for finite element definition. Finally, the reconstructed SPECT data are used as a priori information within a Bayesian reconstruction framework for optical (FMT) reconstruction. We provide results of this multi-modal approach using phantom experimental data and illustrate that this strategy does suppress artifacts and facilitates quantitative analysis for optical imaging studies.

Geometrical co-calibration of a tomographic optical system with CT for intrinsically co-registered imaging

A mathematical approach for geometric co-calibration of a dual-modal small-animal imaging system is presented. The system comprises an optical imaging setup for in vivo bioluminescence and fluorescence detection, as well as an x-ray CT, both mounted on a common rotatable gantry enabling fully simultaneous imaging at axially overlapping fields-of-view. Geometric co-calibration is performed once by imaging a single cylindrical light-emitting source with both modalities over 360 degrees at two axial positions, respectively. Given the three-dimensional coordinates of the source positions in the reconstructed CT volume data along with their two-dimensional locations projected at the optical detector plane, the following intrinsic system parameters are calculated: (i) the intrinsic geometric parameters of the optical detection system-five parameters for each view and (ii) the relative positional relationship between the optical and CT systems-two parameters for each view. After co-calibration is performed, experimental studies using phantoms demonstrate the high degree of intrinsic positional accuracy between the optical and CT measurements. The most important advantage of this approach is that dual-modal data fusion is accomplished without any post-registration strategies.

List-mode maximum-likelihood reconstruction for the ClearPEM system

A dedicated implementation of list-mode maximum-likelihood expectation-maximization (MLEM) reconstruction for the ClearPEM system is presented. The system is composed of two face-to-face detectors, which can be rotated to acquire data from different angular positions. Due to the specific design with irregular sampling and depth of interaction capability, the possible number of lines of response (LOR) is significantly greater than the number of detected events in a standard clinical study. Because reconstruction methods based on data histogramming to sinogram lead to a high computational cost and/or a loss of the intrinsical system resolution, it is necessary to consider the processing of events in list-mode during the reconstruction. The presented method adopted EM algorithm to maximize the logarithmic likelihood function that is expressed in list-mode. The voxel efficiency is corrected by pre-calculated efficiency maps based on flood phantom acquisitions. The method is also implemented with parallelization by distributing the calculation of the acquired events into different threads for significantly increasing computational speed. The results of a Derenzo phantom study show that the presented algorithm can achieve a similar result as 3D-OSEM reconstruction based on data histogramming with significantly lower reconstruction time (6 times faster with one thread, 20 times faster with 8 threads distributed in 8 CPU cores). In clinical studies with lower acquired events, the acceleration ratio can be even higher. The result from a breast phantom study shows that lesions with 15 mm in diameter, each, as well as a small lesion with 5 mm in diameter are clearly visible and can be characterized. The mouse imaging studies show also great potential of the system in small animal applications.

Osteoporosis influences osteogenic but not angiogenic response during bone defect healing in a rat model

Angiogenesis is pivotal for bone metabolism and bone defect healing. Yet the role of vascularization in osteoporosis and osteoporotic bone repair mechanisms is unclear. Here we investigated effects of osteoporotic phenotype on vascularization during bone defect healing in a rodent osteotomy model using volumetric computed tomography (VCT), dynamic contrast-enhanced VCT (DCE-VCT), dynamic contrast-enhanced MRI (DCE-MRI) and histology. In 16 rats, 8 with physiological bone status (SHAM) and 8 with osteoporotic bone status induced by ovariectomy (OVX) in combination with a vitamin D- and low calcium diet, wedge-shaped defects were created at the left distal femur and stabilized internally by T-shaped miniplate. MRI and VCT were performed in all animals 6 weeks after this procedure. By VCT, relative bone density in the defect was evaluated. Using DCE-VCT and DCE-MRI, parameters associated with regional blood volume were calculated in the bone defect, vicinity of the defect, surrounding muscles and bone marrow: Amplitude A and exchange rate constant Kep (DCE-MRI, respectively) as well as peak enhancement PE and area under the curve AUC (DCE-VCT, respectively). In animals of osteoporotic phenotype, bone density within the osseous defect was significantly reduced as compared to SHAM rats. Vascularization parameters determined by DCE-MRI and DCE-VCT in the defect were significantly elevated compared to the adjacent tissues for both SHAM and OVX groups. However, comparing SHAM and OVX rats, no statistically different values were found by DCE-MRI and DCE-VCT concerning any determined vascularization parameter within the bone defect. Furthermore, parameters of vascularization were increased for OVX as compared to SHAM rats within the bone marrow although significant difference was only found for A. In a rat osteotomy model we showed that at the reparative healing stage, osteoporotic phenotype did influence osteogenic but not angiogenic response within bone defect as imaged by DCE-MRI and DCE-VCT. This study provides insight into the relationship between angiogenesis and osteogenesis during osteoporosis-related compromised bone healing.

Quantitative assessment of microcirculation and diffusion in the bone marrow of osteoporotic rats using VCT, DCE-MRI, DW-MRI, and histology

Background Etiologic and pathophysiologic role of functional bone marrow processes is not fully understood especially in the case of osteoporosis. Purpose To investigate the role of vascularization and diffusion in rat models of osteoporosis through a cross-correlation between non-invasive in-vivo imaging and invasive ex-vivo imaging of bone, bone marrow, and in particular of microcirculation. Material and Methods Osteoporosis was induced in rats by combining ovariectomy (OVX) with calcium and Vitamin D3 deficiency, or with glucocorticoid (dexamethasone). For comparison, controls underwent a sham surgery. In in-vivo investigations, animals (n = 36) were examined by volumetric CT (VCT) and MRI at 1, 3, or 12 months post surgery. Using VCT, bone morphology was monitored and relative bone density r within pelvis was extracted. With DCE-MRI and DW-MRI, parameters A (amplitude), Kep (exchange rate constant), and ADC (apparent diffusion coefficient) were acquired for regions of lumbar vertebrae, pelvis, and femur. In ex-vivo investigations, selective histological sections of pelvis were either stained with hematoxylin and eosin (HE stain) for quantifying vessel size and density or immunostained for collagen IV and a-smooth muscle actin to assess vessel maturity (SMA/collagen IV ratio). Results After 12 months, decrease in DCE-MRI parameter Kep was found in all locations of osteoporotic rats (strongest in femur and lumbar vertebrae) while no significant differences were seen for parameter A and DW-MRI parameter ADC. Furthermore, vessel rarefication and maturation were observed on the histological level in animals with osteoporotic phenotype. In particular in the pelvis, the osteoporotic individuals (irrespective of the osteoporosis inducers applied) exhibited decreased Kep, significantly reduced vessel density, significantly increased vessel maturity, as well as statistically unaltered A, ADC, and vessel diameter. Conclusion Changes in microcirculation but not diffusion in the bone marrow of osteoporotic rats are detected by DCE-MRI and DW-MRI due to vessel rarefication and maturation.

Geometric Co-Calibration of an Integrated Small Animal SPECT-CT System for Intrinsically Co-Registered Imaging

We present a method for geometric co-calibration of a single-gantry pinhole SPECT-CT small animal imaging system with axially overlapping fields-of-view. Because spatial resolution and noise level differ greatly between both sub-systems the method incorporates different strategies to obtain more accurate calibration values. Firstly, x-ray tomographic calibration is performed on a steel-sphere-phantom which includes multiple steel spheres with 1 mm in diameter, placed in alignment with a straight line. Geometric displacement for the high-resolution x-ray CT system (five parameters) is then calculated based on a mathematical analysis of those projection data. Secondly, a 360-degree pinhole SPECT-CT imaging of point sources, which are labeled with both radioactivity and x-ray absorber, is applied. With the calculated geometric parameters for the CT system the exact positions of the point sources in the CT volume can be extracted from the CT reconstruction result. Given this positional information as prior the SPECT camera geometric displacement as well as the intrinsic co-alignment between the SPECT and the CT sub-systems (six parameters) are estimated with an iterative optimization strategy (Levenberg-Marquardt). For routine imaging, the calibrated parameters are incorporated into the image reconstruction algorithms applied to the SPECT (MLEM) and CT (FDK) data, respectively, without an additional post-reconstruction registration process. Our results show intrinsically aligned images of both modalities with high spatial resolution and low level misalignment.

Iterative reconstruction of projection images from a microlens-based optical detector

A microlens-based optical detector was developed to perform small animal optical imaging. In this paper we present an iterative reconstruction algorithm yielding improved image quality and spatial resolution as compared to conventional inverse mapping. The reconstruction method utilizes the compressive sensing concept to cope with the undersampling nature of the problem. Each iteration in the algorithm contains two separate steps to ensure both the convergence of the least-square solution and the minimization of the l(1)-norm of the sparsifying transform. The results estimated from measurements, employing a Derenzo-like pattern and a Siemens star phantom, illustrate significant improvements in contrast and spatial resolution in comparison to results calculated by inverse mapping.

Investigating line- versus point-laser excitation for three-dimensional fluorescence imaging and tomography employing a trimodal imaging system

Abstract. The adoption of axially oriented line illumination patterns for fluorescence excitation in small animals for fluorescence surface imaging (FSI) and fluorescence optical tomography (FOT) is being investigated. A trimodal single-photon-emission-computed-tomography/computed-tomography/optical-tomography (SPECT-CT-OT) small animal imaging system is being modified for employment of point- and line-laser excitation sources. These sources can be arbitrarily positioned around the imaged object. The line source is set to illuminate the object along its entire axial direction. Comparative evaluation of point and line illumination patterns for FSI and FOT is provided involving phantom as well as mouse data. Given the trimodal setup, CT data are used to guide the optical approaches by providing boundary information. Furthermore, FOT results are also being compared to SPECT. Results show that line-laser illumination yields a larger axial field of view (FOV) in FSI mode, hence faster data acquisition, and practically acceptable FOT reconstruction throughout the whole animal. Also, superimposed SPECT and FOT data provide additional information on similarities as well as differences in the distribution and uptake of both probe types. Fused CT data enhance further the anatomical localization of the tracer distribution in vivo. The feasibility of line-laser excitation for three-dimensional fluorescence imaging and tomography is demonstrated for initiating further research, however, not with the intention to replace one by the other.

Random correction method for positron emission mammography using delayed coincidence data

A dedicated random correction algorithm is presented in this study for positron emission mammography (PEM) systems. PEM refers to a specified PET system that is optimized for breast imaging by its small FOV. Clinical imaging results from such systems, however, may be degraded by strong statistical noise caused from random coincidences, especially in the region that is near to the torso, due to the high amount of activity uptake outside the FOV, the low geometrical sensitivity of the detector elements near to the torso, and the large solid angle acceptance of random coincidence events. Because of the low statistics of detected coincidence events against the extremely high number of LORs, list-mode reconstruction algorithms are suggested for PEM systems. The conventional random correction methods cannot be directly implemented or can induce an even higher statistical noise. The correction method by single count rate requires a high hardware cost to record single events and needs an accurate calibration to reach a non-bias correction. The new random correction algorithm presented in this study can be implemented into list-mode reconstruction without single count acquisition. This algorithm estimates in a first step a smooth correction image with the delayed coincidences data. This correction image is then used to estimate the mean random coincidence rate for each detected event during the iterative list-mode reconstruction routine. The approach is tested on a ClearPEM system developed by the Crystal Clear Collaboration. Experimental data are acquired by two face-to-face detectors at four angular positions with a total acquisition time of 20 min for each breast. Results show that the proposed algorithm can largely suppress the statistical noise in the region near the torso.

RANGE: Gene Transfer of Reversibly Controlled Polycistronic Genes

We developed a single vector recombinant adeno-associated viral (rAAV) expression system for spatial and reversible control of polycistronic gene expression. Our approach (i) integrates the advantages of the tetracycline (Tet)-controlled transcriptional silencer tTSKid and the self-cleaving 2A peptide bridge, (ii) combines essential regulatory components as an autoregulatory loop, (iii) simplifies the gene delivery scheme, and (iv) regulates multiple genes in a synchronized manner. Controlled by an upstream Tet-responsive element (TRE), both the ubiquitous chicken β-actin promoter (CAG) and the neuron-specific synapsin-1 promoter (Syn) could regulate expression of tTSKid together with two 2A-linked reporter genes. Transduction in vitro exhibited maximally 50-fold regulation by doxycycline (Dox). Determined by gene delivery method as well as promoter, highly specific tissues were transduced in vivo. Bioluminescence imaging (BLI) visualized reversible “ON/OFF” gene switches over repeated “Doxy-Cycling” in living mice. Thus, the reversible rAAV-mediated N-cistronic gene expression system, termed RANGE, may serve as a versatile tool to achieve reversible polycistronic gene regulation for the study of gene function as well as gene therapy.