Free-breathing 3D diffusion MRI for high-resolution hepatic metastasis characterization in small animals

The goal of this study was to develop a 3D diffusion weighted sequence for free breathing liver imaging in small animals at high magnetic field. Hepatic metastases were detected and the apparent diffusion coefficients (ADC) were measured. A 3D SE-EPI sequence was developed by (i) inserting a water-selective excitation radiofrequency pulse to suppress adipose tissue signal and (ii) bipolar diffusion gradients to decrease the sensitivity to respiration motion. Mice with hepatic metastases were imaged at 7T by applying b values from 200 to 1100 s/mm². 3D images with high spatial resolution (182 × 156 × 125 µm) were obtained in only 8 min 32 s. The modified DW-SE-EPI sequence allowed to obtain 3D abdominal images of healthy mice with fat SNR 2.5 times lower than without any fat suppression method and sharpness 2.8 times higher than on respiration-triggered images. Due to the high spatial resolution, the core and the periphery of disseminated hepatic metastases were differentiated at high b-values only, demonstrating the presence of edema and proliferating cells (with ADC of 2.65 × 10^?3 and 1.55 × 10^?3 mm²/s, respectively). Furthermore, these metastases were accurately distinguished from proliferating ones within the same animal at high b-values (mean ADC of 0.38 × 10^?3 mm²/s). Metastases of less than 1.7 mm³ diameter were detected. The new 3D SE-EPI sequence enabled to obtain diffusion information within liver metastases. In addition of intra-metastasis heterogeneity, differences in diffusion were measured between metastases within an animal. This sequence could be used to obtain diffusion information at high magnetic field.     

Calorimetry system for studying small animals following injury

A small-animal calorimetry system designed to study the effect of burn injuries in rats is described. Measurement of sensible and insensible heat loss from single animals can be made for extended periods, while close control of environmental temperatures and humidity is maintained within the calorimeter gradient layer box. Instrumentation, calibration and an improved method of data processing are presented.     

Functional imaging of cerebrovascular activities in small animals using high-resolution photoacoustic tomography

Photoacoustic imaging (PAI) is a noninvasive, nonionizing modality based on the differences in light absorption of various biological tissues. PAI utilizes the endogenous contrast characteristics of traditional optical imaging, while benefiting from high spatial resolution of the ultrasound imaging. A PAI system was developed to reconstruct the two-dimensional cross section image and to visualize the cerebrovascular activities of mouse in vivo. The spatial resolution of the PAI system was determined to be 0.110 mm by a two-point-source phantom with the Rayleigh criterion. The potential applications of the system were clearly demonstrated by successfully mapping a traumatic lesion in the mouse brain cerebral cortex, by its ability to monitor physiological changes in the brain due to carotid ligation and drug stimulation, and two-dimensional sliced images of a traumatic mouse brain at different depths were also provided. Our experimental results indicate that PAI has the potential for studying of traumatic brain injury and physiological functions of the brain.     

Development of high-resolution 4D in vivo-CT for visualization of cardiac and respiratory deformations of small animals

The interest in small animal models of human diseases has generated a need to design a computed tomography (CT) system that operates at a microscopic level. It is particularly important to be able to visualize the dramatic rhythmical motion of organs such as the heart and lungs. In order to evaluate the motion of the heart and lungs of small animals (rats and mice), we developed in the present study a high-resolution 4D in vivo-CT system for small animals that uses synchrotron radiation. To reduce motion artifacts and the radiation dose, the projections were synchronized with airway pressure, the ECG, the x-ray shutter and the CCD shutter. For cardiovascular imaging, a blood pool contrast agent was injected and the data sets were acquired at several ECG points during the end-expiratory phase. For imaging of the lungs, the data sets were acquired at several airway pressures during diastole. The dynamic motion of the cardiovascular system (the ventricles and coronary arteries) and small airways (diameter > 250 microm of rats and 125 microm of mice) was visualized. This high-resolution imaging tool may be very useful for the development of novel drugs in murine models, in addition to its use in the study of cardiovascular and respiratory physiology.     

A telemetry system for recording mastication in small animals

A telemetry system is described for detecting and recording chewing behavior in small laboratory animals. A small pressure sensitive coil implanted in the animal's jaw was used to modulate a small transmitter mounted on the subject's skull. The transmitted signal was received and converted to a d.c. voltage which varies at a rate proportional to the subject's chewing movements. The system may be used to investigate chewing behavior over periods of months.     

A system for the study of behavioral thermoregulation of small animals

A system suitable for the simultaneous monitoring of two unrestrained animals in separate thermoclines is described. It makes use of low-cost, field-proven components to: provide a graded convective and/or conductive temperature continuum over a relatively short runway; measure ambient temperatures at multiple locations within the thermocline; measure the body temperature of a free-running animal within the thermocline; localize the position of the animal within the thermal gradient; and print all the data at periodic intervals. Studies comparing the choices of thermal preference by guinea pigs under diverse conditions indicate that a satisfactory degree of resolution can be achieved by this system.     

Design and performance of a multi-pinhole collimation device for small animal imaging with clinical SPECT and SPECT-CT scanners

A multi-pinhole collimation device is developed that uses the gamma camera detectors of a clinical SPECT or SPECT-CT scanner to produce high-resolution SPECT images. The device consists of a rotating cylindrical collimator having 22 tungsten pinholes with 0.9 mm diameter apertures and an animal bed inside the collimator that moves linearly to provide helical or ordered-subsets axial sampling. CT images also may be acquired on a SPECT-CT scanner for purposes of image co-registration and SPECT attenuation correction. The device is placed on the patient table of the scanner without attaching to the detectors or scanner gantry. The system geometry is calibrated in-place from point source data and is then used during image reconstruction. The SPECT imaging performance of the device is evaluated with test phantom scans. Spatial resolution from reconstructed point source images is measured to be 0.6 mm full width at half maximum or better. Micro-Derenzo phantom images demonstrate the ability to resolve 0.7 mm diameter rod patterns. The axial slabs of a Micro-Defrise phantom are visualized well. Collimator efficiency exceeds 0.05% at the center of the field of view, and images of a uniform phantom show acceptable uniformity and minimal artifact. The overall simplicity and relatively good imaging performance of the device make it an interesting low-cost alternative to dedicated small animal scanners.     

Design and evaluation of a variable aperture collimator for conformal radiotherapy of small animals using a microCT scanner

Treatment of small animals with radiation has in general been limited to planar fields shaped with lead blocks, complicating spatial localization of dose and treatment of deep-seated targets. In order to advance laboratory radiotherapy toward what is accomplished in the clinic, we have constructed a variable aperture collimator for use in shaping the beam of microCT scanner. This unit can image small animal subjects at high resolution, and is capable of delivering therapeutic doses in reasonable exposure times. The proposed collimator consists of two stages, each containing six trapezoidal brass blocks that move along a frame in a manner similar to a camera iris producing a hexagonal aperture of variable size. The two stages are offset by 30 degrees and adjusted for the divergence of the x-ray beam so as to produce a dodecagonal profile at isocenter. Slotted rotating driving plates are used to apply force to pins in the collimator blocks and effect collimator motion. This device has been investigated through both simulation and measurement. The collimator aperture size varied from 0 to 8.5 cm as the driving plate angle increased from 0 to 41 degrees. The torque required to adjust the collimator varied from 0.5 to 5 N x m, increasing with increasing driving plate angle. The transmission profiles produced by the scanner at isocenter exhibited a penumbra of approximately 10% of the collimator aperture width. Misalignment between the collimator assembly and the x-ray source could be identified on the transmission images and corrected by adjustment of the collimator location. This variable aperture collimator technology is therefore a feasible and flexible solution for adjustable shaping of radiation beams for use in small animal radiotherapy as well as other applications in which beam shaping is desired.     

Design of a very high-resolution small animal PET scanner using a silicon scatter detector insert

A small animal positron emission tomography (PET) instrument using a high-resolution solid-state detector insert in a conventional PET system was investigated for its potential to achieve sub-millimeter spatial resolution for mouse imaging. Monte Carlo simulations were used to estimate the effect of detector configurations (thickness, length and radius) on sensitivity. From this initial study, a PET system having an inner cylindrical silicon detector (4 cm ID, 4 cm length and 1.6 cm thickness composed of 16 layers of 300 microm x 300 microm x 1 mm pads), for scattering, surrounded by an outer cylindrical BGO scintillation detector (17.6 cm ID, 16 cm length and 2 cm thickness segmented into 3 mm x 3 mm x 20 mm crystals), for capture was evaluated in detail. In order to evaluate spatial resolution, sensitivity and image quality of the PET system, 2D images of multiple point and cylinder sources were reconstructed with the simulation data including blurring from positron range and annihilation photon acollinearity using filtered backprojection (FBP). Simulation results for (18)F demonstrate 340 microm FWHM at the center of the field of view with 1.0% sensitivity from the coincidence of single scattering events in both silicon detectors and 1.0 mm FWHM with 9.0% sensitivity from the coincidence of single scattering in the silicon and full energy absorption of the second photon in the BGO detector.     

An implantable MEMS micropump system for drug delivery in small animals

We present the first implantable drug delivery system for controlled timing and location of dosing in small animals. Current implantable drug delivery devices do not provide control over these factors nor are they feasible for implantation in research animals as small as mice. Our system utilizes an integrated electrolysis micropump, is refillable, has an inert drug reservoir for broad drug compatibility, and is capable of adjustment to the delivery regimen while implanted. Electrochemical impedance spectroscopy (EIS) was used for characterization of electrodes on glass substrate and a flexible Parylene substrate. Benchtop testing of the electrolysis actuator resulted in flow rates from 1?μL/min to 34?μL/min on glass substrate and up to 6.8?μL/min on Parylene substrate. The fully integrated system generated a flow rate of 4.72?±?0.35?μL/min under applied constant current of 1.0?mA while maintaining a power consumption of only ~3?mW. Finally, we demonstrated in vivo application of the system for anti-cancer drug delivery in mice.     

Activity monitor for small animals

A photoelectric activity monitor which is easily moveable between cages is described, including a circuit diagram and set-up procedure. In a preliminary study, the number of squares traversed by mice on the cage floor was counted both instrumentally and visually; the instrument count ranged from 105 to 111 per 100 observer counts. In a correlation study, a simulated mouse was moved from square to square both laterally and diagonally while being counted by the instrument and a human observer. They agreed 994 and 977 times respectively in 1000 trials. The instrument should prove particularly useful in studying hyperactive mice, whose activity level is too high for the human observer to count accurately. Behavior monitor patterns may also be studied by feeding the channel outputs to a minicomputer.     

A multipinhole small animal SPECT system with submillimeter spatial resolution

Single photon emission computed tomography (SPECT) is an important technology for molecular imaging studies of small animals. In this arena, there is an increasing demand for high performance imaging systems that offer improved spatial resolution and detection efficiency. We have designed a multipinhole small animal imaging system based on position sensitive avalanche photodiode (PSAPD) detectors with the goal of submillimeter spatial resolution and high detection efficiency, which will allow us to minimize the radiation dose to the animal and to shorten the time needed for the imaging study. Our design will use 8 x 24 mm2 PSAPD detector modules coupled to thallium-doped cesium iodide [CsI(Tl)] scintillators, which can achieve an intrinsic spatial resolution of 0.5 mm at 140 keV. These detectors will be arranged in rings of 24 modules each; the animal is positioned in the center of the 9 stationary detector rings which capture projection data from the animal with a cylindrical tungsten multipinhole collimator. The animal is supported on a bed which can be rocked about the central axis to increase angular sampling of the object. In contrast to conventional SPECT pinhole systems, in our design each pinhole views only a portion of the object. However, the ensemble of projection data from all of the multipinhole detectors provide angular sampling that is sufficient to reconstruct tomographic data from the object. The performance of this multipinhole PSAPD imaging system was simulated using a ray tracing program that models the appropriate point spread functions and then was compared against the performance of a dual-headed pinhole SPECT system. The detection efficiency of both systems was simulated and projection data of a hot rod phantom were generated and reconstructed to assess spatial resolution. Appropriate Poisson noise was added to the data to simulate an acquisition time of 15 min and an activity of 18.5 MBq distributed in the phantom. Both sets of data were reconstructed with an ML-EM reconstruction algorithm. In addition, the imaging performance of both systems was evaluated with a uniformity phantom and a realistic digital mouse phantom. Simulations show that our proposed system produces a spatial resolution of 0.8 mm and an average detection efficiency of 630 cps/MBq. In contrast, simulations of the dual-headed pinhole SPECT system produce a spatial resolution of 1.1 mm and an average detection efficiency of 53 cps/MBq. These results suggest that our novel design will achieve high spatial resolution and will improve the detection efficiency by more than an order of magnitude compared to a dual-headed pinhole SPECT system. We expect that this system can perform SPECT with submillimeter spatial resolution, high throughput, and low radiation dose suitable for in vivo imaging of small animals.     

Assessment of the sources of error affecting the quantitative accuracy of SPECT imaging in small animals

Small animal SPECT imaging systems have multiple potential applications in biomedical research. Whereas SPECT data are commonly interpreted qualitatively in a clinical setting, the ability to accurately quantify measurements will increase the utility of the SPECT data for laboratory measurements involving small animals. In this work, we assess the effect of photon attenuation, scatter and partial volume errors on the quantitative accuracy of small animal SPECT measurements, first with Monte Carlo simulation and then confirmed with experimental measurements. The simulations modeled the imaging geometry of a commercially available small animal SPECT system. We simulated the imaging of a radioactive source within a cylinder of water, and reconstructed the projection data using iterative reconstruction algorithms. The size of the source and the size of the surrounding cylinder were varied to evaluate the effects of photon attenuation and scatter on quantitative accuracy. We found that photon attenuation can reduce the measured concentration of radioactivity in a volume of interest in the center of a rat-sized cylinder of water by up to 50% when imaging with iodine-125, and up to 25% when imaging with technetium-99m. When imaging with iodine-125, the scatter-to-primary ratio can reach up to approximately 30%, and can cause overestimation of the radioactivity concentration when reconstructing data with attenuation correction. We varied the size of the source to evaluate partial volume errors, which we found to be a strong function of the size of the volume of interest and the spatial resolution. These errors can result in large (>50%) changes in the measured amount of radioactivity. The simulation results were compared with and found to agree with experimental measurements. The inclusion of attenuation correction in the reconstruction algorithm improved quantitative accuracy. We also found that an improvement of the spatial resolution through the use of resolution recovery techniques (i.e. modeling the finite collimator spatial resolution in iterative reconstruction algorithms) can significantly reduce the partial volume errors.     

MR-based keyhole SPECT for small animal imaging

The rationale for multi-modality imaging is to integrate the strengths of different imaging technologies while reducing the shortcomings of an individual modality. The work presented here proposes a limited-field-of-view (LFOV) SPECT reconstruction technique that can be implemented on a multi-modality MR/SPECT system that can be used to obtain simultaneous MRI and SPECT images for small animal imaging. The reason for using a combined MR/SPECT system in this work is to eliminate any possible misregistration between the two sets of images when MR images are used as a priori information for SPECT. In nuclear imaging the target area is usually smaller than the entire object; thus, focusing the detector on the LFOV results in various advantages including the use of a smaller nuclear detector (less cost), smaller reconstruction region (faster reconstruction) and higher spatial resolution when used in conjunction with pinhole collimators with magnification. The MR/SPECT system can be used to choose a region of interest (ROI) for SPECT. A priori information obtained by the full field-of-view (FOV) MRI combined with the preliminary SPECT image can be used to reduce the dimensions of the SPECT reconstruction by limiting the computation to the smaller FOV while reducing artifacts resulting from the truncated data. Since the technique is based on SPECT imaging within the LFOV it will be called the keyhole SPECT (K-SPECT) method. At first MRI images of the entire object using a larger FOV are obtained to determine the location of the ROI covering the target organ. Once the ROI is determined, the animal is moved inside the radiofrequency (rf) coil to bring the target area inside the LFOV and then simultaneous MRI and SPECT are performed. The spatial resolution of the SPECT image is improved by employing a pinhole collimator with magnification >1 by having carefully calculated acceptance angles for each pinhole to avoid multiplexing. In our design all the pinholes are focused to the center of the LFOV. K-SPECT reconstruction is accomplished by generating an adaptive weighting matrix using a priori information obtained by simultaneously acquired MR images and the radioactivity distribution obtained from the ROI region of the SPECT image that is reconstructed without any a priori input. Preliminary results using simulations with numerical phantoms show that the image resolution of the SPECT image within the LFOV is improved while minimizing artifacts arising from parts of the object outside the LFOV due to the chosen magnification and the new reconstruction technique. The root-mean-square-error (RMSE) in the out-of-field artifacts was reduced by 60% for spherical phantoms using the K-SPECT reconstruction technique and by 48.5-52.6% for the heart in the case with the MOBY phantom. The K-SPECT reconstruction technique significantly improved the spatial resolution and quantification while reducing artifacts from the contributions outside the LFOV as well as reducing the dimension of the reconstruction matrix.     

A simple machine vision-driven system for measuring optokinetic reflex in small animals

The optokinetic reflex (OKR) is useful to monitor the function of the visual and motor nervous systems. However, OKR measurement is not open to all because dedicated commercial equipment or detailed instructions for building in-house equipment is rarely offered. Here we describe the design of an easy-to-install/use yet reliable OKR measuring system including a computer program to visually locate the pupil and a mathematical procedure to estimate the pupil azimuth from the location data. The pupil locating program was created on a low-cost machine vision development platform, whose graphical user interface allows one to compose and operate the program without programming expertise. Our system located mouse pupils at a high success rate (~90 %), estimated their azimuth precisely (~94 %), and detected changes in OKR gain due to the pharmacological modulation of the cerebellar flocculi. The system would promote behavioral assessment in physiology, pharmacology, and genetics.