Discrepancies in stereotaxic coordinate publications and improving precision using an animal-specific atlas.

Rodent brain atlases have traditionally been used to identify brain structures in three-dimensional space for a variety of stereotaxic procedures. As neuroscience becomes increasingly sophisticated, higher levels of precision and consistency are needed. Observations of various atlases currently in use across labs reveal numerous coordinate discrepancies. Here we provide examples of inconsistencies by comparing the coordinates of the boundaries of various brain structures across six atlas publications. We conclude that the coordinates determined by any particular atlas should be considered as only a first approximation of the actual target coordinates for the experimental animal for a particular study. Furthermore, the coordinates determined by one research team cannot be assumed to be universally applicable and accurate in other experimental settings. To optimize precision, we describe a simple protocol for the construction of a customized atlas that is specific to the surgical approach and to the species, gender, and age of the animal used in any given study.

Imaging diagnosis-paraganglioma of the cauda equina: MR findings.

The magnetic resonance imaging features of a cauda equina paraganglioma in a 5-year-old dog are described. Imaging revealed a well-defined, strongly contrast-enhancing mass invading the adjacent vertebral body and infiltrating the intervertebral foramen bilaterally. Flow void, compatible with increased drainage veins around the mass, and macroscopically visible neovessels in contrast-enhanced magnetic resonance angiography, as reported in numerous human studies, were not visible in this single case. The tumor recurred despite aggressive surgery and radiotherapy. This neoplasm should be considered in the differential diagnosis of cauda equina abnormalities in dogs.

Appearance of the canine meninges in subtraction magnetic resonance images.

The canine meninges are not visible as discrete structures in noncontrast magnetic resonance (MR) images, and are incompletely visualized in T1-weighted, postgadolinium images, reportedly appearing as short, thin curvilinear segments with minimal enhancement. Subtraction imaging facilitates detection of enhancement of tissues, hence may increase the conspicuity of meninges. The aim of the present study was to describe qualitatively the appearance of canine meninges in subtraction MR images obtained using a dynamic technique. Images were reviewed of 10 consecutive dogs that had dynamic pre- and postgadolinium T1W imaging of the brain that was interpreted as normal, and had normal cerebrospinal fluid. Image-anatomic correlation was facilitated by dissection and histologic examination of two canine cadavers. Meningeal enhancement was relatively inconspicuous in postgadolinium T1-weighted images, but was clearly visible in subtraction images of all dogs. Enhancement was visible as faint, small-rounded foci compatible with vessels seen end on within the sulci, a series of larger rounded foci compatible with vessels of variable caliber on the dorsal aspect of the cerebral cortex, and a continuous thin zone of moderate enhancement around the brain. Superimposition of color-encoded subtraction images on pregadolinium T1- and T2-weighted images facilitated localization of the origin of enhancement, which appeared to be predominantly dural, with relatively few leptomeningeal structures visible. Dynamic subtraction MR imaging should be considered for inclusion in clinical brain MR protocols because of the possibility that its use may increase sensitivity for lesions affecting the meninges.

Individual nodule tracking in micro-CT images of a longitudinal lung cancer mouse model.

We present and evaluate an automatic and quantitative method for the complex task of characterizing individual nodule volumetric progression in a longitudinal mouse model of lung cancer. Fourteen A/J mice received an intraperitoneal injection of urethane. Respiratory-gated micro-CT images of the lungs were acquired at 8, 22, and 37 weeks after injection. A radiologist identified a total of 196, 585 and 636 nodules, respectively. The three micro-CT image volumes from every animal were then registered and the nodules automatically matched with an average accuracy of 99.5%. All nodules detected at week 8 were tracked all the way to week 37, and volumetrically segmented to measure their growth and doubling rates. 92.5% of all nodules were correctly segmented, ranging from the earliest stage to advanced stage, where nodule segmentation becomes more challenging due to complex anatomy and nodule overlap. Volume segmentation was validated using a foam lung phantom with embedded polyethylene microspheres. We also correlated growth rates with nodule phenotypes based on histology, to conclude that the growth rate of malignant tumors is significantly higher than that of benign lesions. In conclusion, we present a turnkey solution that combines longitudinal imaging with nodule matching and volumetric nodule segmentation resulting in a powerful tool for preclinical research.

Canine meningeal disease: associations between magnetic resonance imaging signs and histologic findings.

In order to compare the accuracy of MR sequences for diagnosis of meningeal disease, MR images of the brain, and histopathologic specimens including the meninges of 60 dogs were reviewed retrospectively by independent observers in a cross-sectional study. MR images included T1-weighted pre- and postgadolinium images, subtraction images, T2-weighted images, and T2-weighted fluid-attenuated inversion-recovery (FLAIR) images. Pathologic changes affected the pachymeninges in 16 dogs, leptomeninges in 35 dogs, and brain in 38 dogs. The meninges were normal in 12 dogs. Meninges were classified histopathologically as normal (grade 0), slightly or inconsistently affected (grade 1), or markedly affected (grade 2). When applying relaxed pathologic criteria (grades 0 and 1 considered normal), the results of ROC analysis (area under curve, AUC) were: T1-weighted postcontrast images 0.74; subtraction images 0.7; T2-weighted images 0.68; FLAIR images 0.56. The difference in AUC between T1-weighted postgadolinium images and FLAIR images was significant (P = 0.04). AUC for FLAIR images was not significantly different from 0.5. When applying strict pathologic criteria (only grade 0 considered normal), none of the MR sequences had AUC significantly different from 0.5. On the basis of T1-weighted postgadolinium images and subtraction images, correct anatomic classification of lesions occurred more often for pachymeningeal than leptomeningeal lesions (P < 0.001). Overall, MR imaging had low sensitivity for diagnosis of meningeal pathology in dogs, particularly for changes affecting the leptomeninges. Subtraction images had similar accuracy to T1-weighted postgadolinium images for meningeal lesions in dogs. T2-weighted FLAIR images appear to have limited diagnostic utility for meningeal lesions.

Registration of challenging pre-clinical brain images.

The size and complexity of brain imaging studies in pre-clinical populations are increasing, and automated image analysis pipelines are urgently required. Pre-clinical populations can be subjected to controlled interventions (e.g., targeted lesions), which significantly change the appearance of the brain obtained by imaging. Existing systems for registration (the systematic alignment of scans into a consistent anatomical coordinate system), which assume image similarity to a reference scan, may fail when applied to these images. However, affine registration is a particularly vital pre-processing step for subsequent image analysis which is assumed to be an effective procedure in recent literature describing sophisticated techniques such as manifold learning. Therefore, in this paper, we present an affine registration solution that uses a graphical model of a population to decompose difficult pairwise registrations into a composition of steps using other members of the population. We developed this methodology in the context of a pre-clinical model of stroke in which large, variable hyper-intense lesions significantly impact registration performance. We tested this technique systematically in a simulated human population of brain tumour images before applying it to pre-clinical models of Parkinson's disease and stroke.

CT-PET weighted image fusion for separately scanned whole body rat.

PURPOSE: The limited resolution and lack of spatial information in positron emission tomography (PET) images require the complementary anatomic information from the computed tomography (CT) and/or magnetic resonance imaging (MRI). Therefore, multimodality image fusion techniques such as PET/CT are critical in mapping the functional images to structural images and thus facilitate the interpretation of PET studies. In our experimental situation, the CT and PET images are acquired in separate scanners at different times and the inherent differences in the imaging protocols produce significant nonrigid changes between the two acquisitions in addition to dissimilar image characteristics. The registration conditions are also poor because CT images have artifacts due to the limitation of current scanning settings, while PET images are very blurry (in transmission-PET) and have vague anatomical structure boundaries (in emission-PET). METHODS: The authors present a new method for whole body small animal multimodal registration. In particular, the authors register whole body rat CT image and PET images using a weighted demons algorithm. The authors use both the transmission-PET and the emission-PET images in the registration process emphasizing particular regions of the moving transmission-PET image using the emission-PET image. After a rigid transformation and a histogram matching between the CT and the transmission-PET images, the authors deformably register the transmission-PET image to the CT image with weights based on the intensity-normalized emission-PET image. For the deformable registration process, the authors develop a weighted demons registration method that can give preferences to particular regions of the input image using a weight image. RESULTS: The authors validate the results with nine rat image sets using the M-Hausdorff distance (M-HD) similarity measure with different outlier-suppression parameters (OSP). In comparison with standard methods such as the regular demons and the normalized mutual information (NMI)-based nonrigid free-form deformation (FFD) registration, the proposed weighted demons registration method shows average M-HD errors: 3.99 ± 1.37 (OSP = 10), 5.04 ± 1.59 (OSP = 20) and 5.92 ± 1.61 (OSP = ∞) with statistical significance (p < 0.0003) respectively, while NMI-based nonrigid FFD has average M-HD errors: 5.74 ± 1.73 (OSP = 10), 7.40 ± 7.84 (OSP = 20) and 9.83 ± 4.13 (OSP = ∞), and the regular demons has average M-HD errors: 6.79 ± 0.83 (OSP = 10), 9.19 ± 2.39 (OSP = 20) and 11.63 ± 3.99 (OSP = ∞), respectively. In addition to M-HD comparisons, the visual comparisons on the faint-edged region between the CT and the aligned PET images also show the encouraging improvements over the other methods. CONCLUSIONS: In the whole body multimodal registration between CT and PET images, the utilization of both the transmission-PET and the emission-PET images in the registration process by emphasizing particular regions of the transmission-PET image using an emission-PET image is effective. This method holds promise for other image fusion applications where multiple (more than two) input images should be registered into a single informative image.

Hybrid small animal imaging system combining magnetic resonance imaging with fluorescence tomography using single photon avalanche diode detectors.

The high sensitivity of fluorescence imaging enables the detection of molecular processes in living organisms. However, diffuse light propagation in tissue prevents accurate recovery of tomographic information on fluorophore distribution for structures embedded deeper than 0.5 mm. Combining optical with magnetic resonance imaging (MRI) provides an accurate anatomical reference for fluorescence imaging data and thereby enables the correlation of molecular with high quality structural/functional information. We describe an integrated system for small animal imaging incorporating a noncontact fluorescence molecular tomography (FMT) system into an MRI detector. By adopting a free laser beam design geometrical constraints imposed by the use of optical fibers could be avoided allowing for flexible fluorescence excitation schemes. Photon detection based on a single-photon avalanche diode array enabled simultaneous FMT/MRI measurements without interference between modalities. In vitro characterization revealed good spatial accuracy of FMT data and accurate quantification of dye concentrations. Feasibility of FMT/MRI was demonstrated in vivo by simultaneous assessment of protease activity and tumor morphology in murine colon cancer xenografts.

Assessment of MR compatibility of a PET insert developed for simultaneous multiparametric PET/MR imaging on an animal system operating at 7 T.

The combination of positron emission tomography and MR in one system is currently emerging and opens up new domains in the functional examinations of living systems. This article reports on relevant influences of a positron emission tomography insert on MR imaging. The basic conditions of main magnetic field and RF field homogeneity were measured as well as image quality and signal-to-noise ratio when applying the usual MR sequence types including echo-planar techniques. Moreover, the influence of the positron emission tomography insert on the RF noise level and on RF interferences was measured by comparing results achieved with and without the positron emission tomography insert. The temporal stability of EPI imaging with and without the positron emission tomography insert was assessed. Small but significant decreases in the signal-to-noise ratio were revealed when the positron emission tomography insert was present, whereas B(0) and B(1) homogeneity as well as RF noise level were not adversely affected. A higher signal intensity drift was found for EPI imaging studies; however, this can be compensated by post processing. In summary, this study shows that positron emission tomography inserts can be designed for and used within an MR system practically, without substantially affecting the MR image quality.

Quantitative fluorescence tomography using a trimodality system: in vivo validation.

A fully integrated trimodality fluorescence, diffuse optical, and x-ray computed tomography (FT/DOT/XCT) system for small animal imaging is reported in this work. The main purpose of this system is to obtain quantitatively accurate fluorescence concentration images using a multimodality approach. XCT offers anatomical information, while DOT provides the necessary background optical property map to improve FT image accuracy. The quantitative accuracy of this trimodality system is demonstrated in vivo. In particular, we show that a 2-mm-diam fluorescence inclusion located 8 mm deep in a nude mouse can only be localized when functional a priori information from DOT is available. However, the error in the recovered fluorophore concentration is nearly 87%. On the other hand, the fluorophore concentration can be accurately recovered within 2% error when both DOT functional and XCT structural a priori information are utilized together to guide and constrain the FT reconstruction algorithm.

Design and performance evaluation of a coplanar multimodality scanner for rodent imaging.

This work reports on the development and performance evaluation of the VrPET/CT, a new multimodality scanner with coplanar geometry for in vivo rodent imaging. The scanner design is based on a partial-ring PET system and a small-animal CT assembled on a rotatory gantry without axial displacement between the geometric centers of both fields of view (FOV). We report on the PET system performance based on the NEMA NU-4 protocol; the performance characteristics of the CT component are not included herein. The accuracy of inter-modality alignment and the imaging capability of the whole system are also evaluated on phantom and animal studies. Tangential spatial resolution of PET images ranged between 1.56 mm at the center of the FOV and 2.46 at a radial offset of 3.5 cm. The radial resolution varies from 1.48 mm to 1.88 mm, and the axial resolution from 2.34 mm to 3.38 mm for the same positions. The energy resolution was 16.5% on average for the entire system. The absolute coincidence sensitivity is 2.2% for a 100-700 keV energy window with a 3.8 ns coincident window. The scatter fraction values for the same settings were 11.45% for a mouse-sized phantom and 23.26% for a rat-sized phantom. The peak noise equivalent count rates were also evaluated for those phantoms obtaining 70.8 kcps at 0.66 MBq/cc and 31.5 kcps at 0.11 MBq/cc, respectively. The accuracy of inter-modality alignment is below half the PET resolution, and the image quality of biological specimens agrees with measured performance parameters. The assessment presented in this study shows that the VrPET/CT system is a good performance small-animal imager, while the cost derived from a partial ring detection system is substantially reduced as compared with a full-ring PET tomograph.

Registration of planar bioluminescence to magnetic resonance and x-ray computed tomography images as a platform for the development of bioluminescence tomography reconstruction algorithms.

The procedures we propose make possible the mapping of two-dimensional (2-D) bioluminescence image (BLI) data onto a skin surface derived from a three-dimensional (3-D) anatomical modality [magnetic resonance (MR) or computed tomography (CT)] dataset. This mapping allows anatomical information to be incorporated into bioluminescence tomography (BLT) reconstruction procedures and, when applied using sources visible to both optical and anatomical modalities, can be used to evaluate the accuracy of those reconstructions. Our procedures, based on immobilization of the animal and a priori determined fixed projective transforms, should be more robust and accurate than previously described efforts, which rely on a poorly constrained retrospectively determined warping of the 3-D anatomical information. Experiments conducted to measure the accuracy of the proposed registration procedure found it to have a mean error of 0.36+/-0.23 mm. Additional experiments highlight some of the confounds that are often overlooked in the BLT reconstruction process, and for two of these confounds, simple corrections are proposed.

A non-rigid registration method for serial microCT mouse hindlimb images.

We present a new method for the non-rigid registration of serial mouse microCT images which undergo potentially large changes in the positions of the legs due to articulation. While non-rigid registration methods have been extensively used in the evaluation of individual organs, application in whole body imaging has been limited, primarily because the scale of possible displacements and deformations is large resulting in poor convergence of most methods. Our method is based on the extended demons algorithm that uses a level-set representation of the mouse skin and skeleton as an input, and composed of three steps reflecting the natural physical movements of bony structures. We applied our method to the registration of serial microCT mouse images demonstrating encouraging performances as compared to competitive techniques.

A registration based approach for 4D cardiac micro-CT using combined prospective and retrospective gating.

Recent advances in murine cardiac studies with three-dimensional cone beam micro-computed tomography (CT) have used either prospective or retrospective gating technique. While prospective gating ensures the best image quality and the highest resolution, it involves longer sampling times and higher radiation dose. Sampling is faster and the radiation dose can be reduced with retrospective gating but the image quality is affected by the limited number of projections with an irregular angular distribution which complicate the reconstruction process, causing significant streaking artifacts. This work involves both prospective and retrospective gating in sampling. Deformable registration is used between a high quality image set acquired with prospective gating with the multiple data sets during the cardiac cycle obtained using retrospective gating. Tests were conducted on a four-dimensional (4D) cardiac mouse phantom and after optimization, the method was applied to in vivo cardiac micro-CT data. Results indicate that, by using our method, the sampling time can be reduced by a factor of 2.5 and the radiation dose can be reduced 35% compared to the prospective sampling while the image quality can be maintained. In conclusion, we proposed a novel solution to 4D cine cardiac micro-CT based on a combined prospective with retrospective gating in sampling and deformable registration post reconstruction that mixed the advantages of both strategies.

Automatic nonrigid registration of whole body CT mice images.

Three-dimensional intra- and intersubject registration of image volumes is important for tasks that include quantification of temporal/longitudinal changes, atlas-based segmentation, computing population averages, or voxel and tensor-based morphometry. While a number of methods have been proposed to address this problem, few have focused on the problem of registering whole body image volumes acquired either from humans or small animals. These image volumes typically contain a large number of articulated structures, which makes registration more difficult than the registration of head images, to which the majority of registration algorithms have been applied. This article presents a new method for the automatic registration of whole body computed tomography (CT) volumes, which consists of two main steps. Skeletons are first brought into approximate correspondence with a robust point-based method. Transformations so obtained are refined with an intensity-based nonrigid registration algorithm that includes spatial adaptation of the transformation's stiffness. The approach has been applied to whole body CT images of mice, to CT images of the human upper torso, and to human head and neck CT images. To validate the authors method on soft tissue structures, which are difficult to see in CT images, the authors use coregistered magnetic resonance images. They demonstrate that the approach they propose can successfully register image volumes even when these volumes are very different in size and shape or if they have been acquired with the subjects in different positions.

Simultaneous in vivo dynamic magnetic resonance-diffuse optical tomography for small animal imaging.

We present simultaneous measurement of enhancement kinetics of an optical and a magnetic resonance (MR) contrast agent in a small animal breast tumor model (R3230 ac) using a combined MR-diffuse optical tomographic (MR-DOT) imaging system. A mixture of a small molecular-weight MR contrast agent gadolinium-diethylene-triamine-pentaacetic acid (Gd-DTPA) and a large molecular-weight optical contrast agent indocyanine green (ICG) was administered intravenously for multimodal dynamic imaging. Coregistration of optical and MR images was accomplished using agar-water-based markers. Using T(2) and dynamic T(1) weighted MR images, we divided the entire tumor into two regions of interest (ROI): a viable and a nonviable region. The absorption enhancements in the ROIs were calculated. An enhancement of the ICG was observed in the viable region. On the contrary, there was a lower enhancement in the nonviable region.

Rodent brain imaging with SPECT/CT.

We evaluated methods of imaging rat models of stroke in vivo using a single photon emission computed tomography (SPECT) system dedicated to small animal imaging (X-SPECT, Gamma Medica-Ideas, Northridge, CA). An animal model of ischemic stroke was developed for in vivo SPECT/CT imaging using the middle cerebral artery occlusion (MCAO) technique. The presence of cerebral ischemia was verified in ex vivo studies using triphenyltetrazolium chloride (TTC) staining. In vivo radionuclide imaging of cerebral blood flow was performed in rats following MCAO using dynamic planar imaging of 99mTc-exametazime with parallel hole collimation. This was followed immediately by in vivo radionuclide imaging of cerebral blood flow with 99mTc-exametazime in the same animals using 1-mm pinhole SPECT. Correlated computed tomography imaging was performed to localize radiopharmaceutical uptake. The animals were allowed to recover and ex vivo autoradiography was performed with separate administration of 99mTc-exametazime. Time activity curve of 99mTc-exametazime showed that the radiopharmaceutical uptake could be maintained for over 9 min. The activity would be expected to be relatively stable for a much longer period, although the data were only obtained for 9 min. TTC staining revealed sizable infarcts by visual observation of inexistence of TTC stain in infracted tissues of MCAO rat brains. In vivo SPECT imaging showed cerebral blood flow deficit in the MCAO model, and the in vivo imaging result was confirmed with ex vivo autoradiography. We have demonstrated a capability of imaging regions of cerebral blood flow deficit in MCAO rat brains in vivo using a pinhole SPECT dedicated to small animal imaging.

Introduction to clinical and laboratory (small-animal) image registration and fusion.

Imaging has long been a vital component of clinical medicine and, increasingly, of biomedical research in small-animals. Clinical and laboratory imaging modalities can be divided into two general categories, structural (or anatomical) and functional (or physiological). The latter, in particular, has spawned what has come to be known as "molecular imaging". Image registration and fusion have rapidly emerged as invaluable components of both clinical and small-animal imaging and has lead to the development and marketing of a variety of multi-modality, e.g. PET-CT, devices which provide registered and fused three-dimensional image sets. This paper briefly reviews the basics of image registration and fusion and available clinical and small-animal multi-modality instrumentation.

Broad-spectrum multi-modality image registration: from PET, CT, and MRI to autoradiography, microscopy, and beyond.

Image registration and fusion are increasingly important components of both clinical and small-animal imaging and have lead to the development of a variety of pertinent hardware and software tools, including multi-modality, e.g. PET-CT, devices. At the same time, advances in microscopic imaging, including phosphor-plate digital autoradiography and immunohistochemistry, now allow ultra-high (sub-100 microm)-resolution molecular characterization of tissue sections. To date, however, in vivo imaging of intact subjects and ex vivo imaging of harvested tissues sections have remained separate and distinct, making it difficult to reliably inter-compare the former and the latter. The Department of Medical Physics and the Radiation Biophysics Laboratory at Memorial Sloan-Kettering Cancer Center, under the direction of Dr. Clifton Ling, has now designed, fabricated, and tested a stereotactic imaging system for so-called "broad-spectrum" image registration, from coarser-resolution in vivo imaging modalities such as PET, CT, and MRI to ultra-high-resolution ex vivo imaging techniques such as histology, autoradiography, and immunohistochemistry.

Differentiation between chondroma and squamous cell carcinoma: evaluation of subtraction image of MRI.

The image processing procedure has become widely applied as a visual aid in imaging diagnosis. The subtraction image of MRI obtained by digitally subtracting an unenhanced image from a contrast-enhanced image, depicted a discrete distribution of the contrast agent. Subtraction images in the clinical cases were compared. The subtraction image in a case of chondroma demonstrated a relatively uniform distribution of a contrast agent, with a well delineated neoplastic lesion. The subtraction image in a case of squamous cell carcinoma, the contrast agent was distributed heterogeneously in the carcinoma and well permeated into the muscle. Different patterns of subtraction image were obtained in the clinical cases of chondroma and squamous cell carcinoma in this study. The findings suggest the potential usefulness of the subtraction image for diagnosing the degree of malignancy.