Performance evaluation of the mouse version of the LabPET II PET scanner.

The LabPET II is a new positron emission tomography technology platform designed to achieve submillimetric spatial resolution imaging using fully pixelated avalanche photodiodes-based detectors and highly integrated parallel front-end processing electronics. The detector was designed as a generic building block to develop devices for preclinical imaging of small to mid-sized animals and for clinical imaging of the human brain. The aim of this work is to assess the physical characteristics and imaging performance of the mouse version of LabPET II scanner following the NEMA NU4-2008 standard and using high resolution phantoms and in vivo imaging applications. A reconstructed spatial resolution of 0.78 mm (0.5 µ l) is measured close to the center of the radial field of view. With an energy window of 350 650 keV, the system absolute sensitivity is 1.2% and its maximum noise equivalent count rate reaches 61.1 kcps at 117 MBq. Submillimetric spatial resolution is achieved in a hot spot phantom and tiny bone structures were resolved with unprecedented contrast in the mouse. These results provide convincing evidence of the capabilities of the LabPET II technology for biomolecular imaging in preclinical research.

Imaging performance of LabPET APD-based digital PET scanners for pre-clinical research.

The LabPET is an avalanche photodiode (APD) based digital PET scanner with quasi-individual detector read-out and highly parallel electronic architecture for high-performance in vivo molecular imaging of small animals. The scanner is based on LYSO and LGSO scintillation crystals (2×2×12/14 mm3), assembled side-by-side in phoswich pairs read out by an APD. High spatial resolution is achieved through the individual and independent read-out of an individual APD detector for recording impinging annihilation photons. The LabPET exists in three versions, LabPET4 (3.75 cm axial length), LabPET8 (7.5 cm axial length) and LabPET12 (11.4 cm axial length). This paper focuses on the systematic characterization of the three LabPET versions using two different energy window settings to implement a high-efficiency mode (250­650 keV) and a high-resolution mode (350­650 keV) in the most suitable operating conditions. Prior to measurements, a global timing alignment of the scanners and optimization of the APD operating bias have been carried out. Characteristics such as spatial resolution, absolute sensitivity, count rate performance and image quality have been thoroughly investigated following the NEMA NU 4-2008 protocol. Phantom and small animal images were acquired to assess the scanners' suitability for the most demanding imaging tasks in preclinical biomedical research. The three systems achieve the same radial FBP spatial resolution at 5 mm from the field-of-view center: 1.65/3.40 mm (FWHM/FWTM) for an energy threshold of 250 keV and 1.51/2.97 mm for an energy threshold of 350 keV. The absolute sensitivity for an energy window of 250­650 keV is 1.4%/2.6%/4.3% for LabPET4/8/12, respectively. The best count rate performance peaking at 362 kcps is achieved by the LabPET12 with an energy window of 250­650 keV and a mouse phantom (2.5 cm diameter) at an activity of 2.4 MBq ml−1. With the same phantom, the scatter fraction for all scanners is about 17% for an energy threshold of 250 keV and 10% for an energy threshold of 350 keV. The results obtained with two energy window settings confirm the relevance of high-efficiency and high-resolution operating modes to take full advantage of the imaging capabilities of the LabPET scanners for molecular imaging applications.

PET-based geometrical calibration of a pinhole SPECT add-on for an animal PET scanner.

We developed SPECT imaging capability on an animal PET scanner to provide a cost effective option for animal SPECT imaging. The SPECT add-on sub-system was enabled by mechanically integrating a multiple-pinhole collimator in the PET detector ring. This study introduces a method to calibrate the geometrical parameters of the SPECT add-on using the PET imaging capability of the scanner. The proposed PET imaging-based calibration method consists of two steps: (1) paint the pinhole apertures of the collimator with a positron emitting radioactive solution; and (2) image the collimator inside the scanner in PET mode. The geometrical parameters of the multi-pinhole SPECT add-on can then be derived directly from a set of PET images by simple linear calculation and used in defining the SPECT system. The method was compared to our implementation of a SPECT calibration approach with model-based fitting of SPECT projection data. The procedure for carrying out the PET imaging-based calibration method is simpler and faster than that of our implementation of the SPECT model-based calibration method. Since it does not require model fitting, the uniqueness of the calibration result is warranted. Better quality SPECT images were reconstructed using the PET-derived calibration parameters rather than our implementation of the SPECT model-based calibration parameters. We conclude that the proposed PET imaging-based calibration method provides a highly effective means for enabling SPECT imaging on a PET scanner.

Mechanism of reduced myocardial glucose utilization during acute hypertriglyceridemia in rats.

PURPOSE: The purpose of the research is to study the effect of acute inhibition of intravascular lipolysis on myocardial substrate selection during hypertriglyceridemia using in vivo radiotracer analysis and positron emission tomography. PROCEDURES: We induced acute hypertriglyceridemia in vivo using an intravenous infusion of Intralipid 20% (IL) without and with acute inhibition of fatty acid delivery from circulating triglycerides with injection of Triton WR-1339 (TRI) during a euglycemic-hyperinsulinemic clamp in Wistar rats. We determined the effect of TRI on myocardial uptake of circulating triglycerides and free fatty acids using intravenous injection of [(3)H]-triolein and [(14)C]-bromopalmitate, respectively. Myocardial blood flow, oxidative metabolism, and metabolic rate of glucose (MMRG) were determined using micro-positron emission tomography (microPET) with [(13)N]-ammonia, [(11)C]-acetate, and 2-deoxy-2-[F-18]fluoro-D: -glucose (FDG). RESULTS: TRI reduced myocardial incorporation of [(3)H]-triolein but not [(14)C]-bromopalmitate showing that it selectively reduces myocardial fatty acid delivery from circulating triglycerides but not from free fatty acids. IL reduced myocardial blood flow and MMRG by 37% and 56%, respectively, but did not affect myocardial oxidative metabolism. TRI did not abolish the effect of IL on myocardial blood flow and MMRG. CONCLUSIONS: Hypertriglyceridemia acutely reduces myocardial blood flow and MMRG in rats, but this effect is not explained by increased myocardial fatty acid delivery through intravascular triglyceride lipolysis.

A new tool for molecular imaging: the microvolumetric beta blood counter.

UNLABELLED: Radiotracer kinetic modeling in small animals with PET allows absolute quantification of physiologic and biochemical processes in vivo. It requires blood and tissue tracer concentrations as a function of time. Manual sampling, the reference method for blood tracer concentration measurements, requires fairly large amounts of blood besides being technically difficult and time-consuming. An automated microvolumetric beta blood counter (microBC) was designed to circumvent these limitations by measuring the blood activity in real time with PET scanning. METHODS: The microBC uses direct beta-particle detection to reduce its footprint and is entirely remote controlled for sampling protocol selection and real-time monitoring of measured parameters. Sensitivity has been determined for the most popular PET radioisotopes ((18)F, (13)N, (11)C, (64)Cu). Dispersion within the sampling catheter has been modeled to enable automatic correction. Blood curves obtained with the microBC were compared with manual samples and PET-derived data. The microBC was used to estimate the myocardial blood flow (MBF) of mice injected with (13)N-ammonia and to compare the myocardial metabolic rate of glucose (MMRG) of rats injected with (18)F-FDG for arterial and venous cannulation sites. RESULTS: The sensitivity limit ranges from 3 to 104 Bq/microL, depending on the isotope and the catheter used, and was found to be adequate for most small-animal studies. Automatic dispersion correction appears to be a good approximation of dispersion-free reference curves. Blood curves sampled with the microBC are well correlated with curves obtained from manual samples and PET images. With correction for dispersion, the MBF of anesthetized mice at rest was found to be 4.84 +/- 0.5 mL/g/min, which is comparable to values found in the literature for rats. MMRG values derived from the venous blood tracer concentration are underestimated by 60% as compared with those derived from arterial blood. CONCLUSION: The microBC is a compact automated counter allowing real-time measurement of blood radioactivity for pharmacokinetic studies in animals as small as mice. Reliable and reproducible, the device makes it possible to increase the throughput of pharmacokinetic studies with reduced blood sample handling and staff exposure, contributing to speed up new drug development and evaluation.

A small animal positron emission tomography study of the effect of chemotherapy and hormonal therapy on the uptake of 2-deoxy-2-[F-18]fluoro-D-glucose in murine models of breast cancer.

PURPOSE: We used small animal positron emission tomography (PET) imaging to monitor the time-course of tumor metabolic response to hormone and chemotherapy in a murine model of hormone-sensitive breast cancer. PROCEDURES: Estrogen receptor positive murine mammary carcinomas were inoculated in Balb/c mice. Small animal PET imaging using 2-deoxy-2-[F-18]fluoro-D: -glucose (FDG) was used to assess tumor metabolic activity. Imaging was done before and at days 1, 7, and 14 after the administration of doxorubicin, methotrexate, letrozole, or placebo. The tumor uptake of FDG was calculated from a region-of-interest drawn around the tumor. RESULTS: All treatments resulted in a decrease in tumor growth rate and end volume compared to untreated control. FDG uptake was also markedly decreased after treatment although a flare reaction was observed on PET at day 7, the intensity of which varied according to the treatment modality. CONCLUSION: PET imaging is sensitive to detect early changes associated with therapy in murine breast cancer models. A flare reaction was observed 7 days after the initiation of therapy.

PET imaging of apoptosis with (64)Cu-labeled streptavidin following pretargeting of phosphatidylserine with biotinylated annexin-V.

PURPOSE: In vivo detection of apoptosis is a diagnostic tool with potential clinical applications in cardiology and oncology. Radiolabeled annexin-V (anxV) is an ideal probe for in vivo apoptosis detection owing to its strong affinity for phosphatidylserine (PS), the molecular flag on the surface of apoptotic cells. Most clinical studies performed to visualize apoptosis have used (99m)Tc-anxV; however, its poor distribution profile often compromises image quality. In this study, tumor apoptosis after therapy was visualized by positron emission tomography (PET) using (64)Cu-labeled streptavidin (SAv), following pre-targeting of apoptotic cells with biotinylated anxV. METHODS: Apoptosis was induced in tumor-bearing mice by photodynamic therapy (PDT) using phthalocyanine dyes as photosensitizers, and red light. After PDT, mice were injected i.v. with biotinylated anxV, followed 2 h later by an avidin chase, and after another 2 h with (64)Cu-DOTA-biotin-SAv. PET images were subsequently recorded up to 13 h after PDT. RESULTS: PET images delineated apoptosis in treated tumors as early as 30 min after (64)Cu-DOTA-biotin-SAv administration, with tumor-to-background ratios reaching a maximum at 3 h post-injection, i.e., 7 h post-PDT. Omitting the administration of biotinylated anxV or the avidin chase failed to provide a clear PET image, confirming that all three steps are essential for adequate visualization of apoptosis. Furthermore, differences in action mechanisms between photosensitizers that target tumor cells directly or via initial vascular stasis were clearly recognized through differences in tracer uptake patterns detecting early or delayed apoptosis. CONCLUSION: This study demonstrates the efficacy of a three-step (64)Cu pretargeting procedure for PET imaging of apoptosis. Our data also confirm the usefulness of small animal PET to evaluate cancer treatment protocols.

Dynamic imaging of transient metabolic processes by small-animal PET for the evaluation of photosensitizers in photodynamic therapy of cancer.

UNLABELLED: This study evaluated the potential use of dynamic PET to monitor transient metabolic processes and to investigate the mechanisms of action of new photosensitizing drugs in the photodynamic therapy (PDT) of cancer. METHODS: Rats bearing 2 mammary tumors received different phthalocyanine-based photosensitizers. The following day, the animals were positioned in a Sherbrooke small-animal PET scanner and continuously infused with 18F-FDG while dynamic images were acquired for 2 h. During that period, one of the 2 tumors was exposed for 30 min to red light delivered by a small diode laser to activate PDT. RESULTS: 18F-FDG time-activity curves during PDT showed distinct transient patterns characterized by a drop and subsequent recovery of tumor 18F-FDG uptake rates. Variations in these rates and response delay parameters revealed tumoral and systemic metabolic processes that correlated with differences in mechanism of action between drugs, that is, direct tumor cell kill or initial vascular shutdown. CONCLUSION: Real-time follow-up of tumor response to PDT as monitored by dynamic 18F-FDG PET has been shown to correlate with the mechanisms of action of photosensitizing drugs in vivo. This new imaging paradigm can be exploited to monitor a variety of transient cellular and molecular processes as they occur in vivo, enabling the mechanisms of action of therapeutic interventions to be scrutinized and their efficacy predicted in real time.

Breast cancer models to study the expression of estrogen receptors with small animal PET imaging.

Different animal models of estrogen positive tumors (ER+) were evaluated for their suitability to follow tumor response after various treatment protocols, using small animal positron emission tomography (PET). ER+ human breast cancer cell lines MCF-7 and T-47D, using MDA-MB-231 as ER-; control, and murine mammary ductal carcinomas MC4-L2, MC4-L3, and MC7-L1, were compared for their in vivo growth rate and retention of ER+ status. Tumor metabolic activity was estimated from the relative uptake (% injected dose/g) of [18F]fluorodeoxyglucose (FDG) uptake, whereas ER content was determined from 16alpha-[18F]fluoroestradiol (FES) retention. F-18 activity values were obtained by small animal PET imaging and confirmed by tissue sampling and radioactivity counting. Reliable uptake measurements could be obtained for tumors of 200 microl or over. The human cell lines grew at a slower rate in vivo and failed to accumulate FES; in contrast, the Balb/c MC7-L1 and MC4-L2 grew well and showed good uptake of both FDG and FES. Chemotherapy and hormone therapy delayed the growth of MC7-L1 and MC4-L2 tumors, confirming their suitability as an ER+ model for therapeutic interventions. MC4-L3 tumors also showed promising results but required the presence of progestative pellets to grow. These data demonstrate that murine MC7-L1 and MC4-L2 tumors are suitable models for the monitoring of ER+ breast cancer therapy using small animal PET imaging.

Quantitative gated PET for the assessment of left ventricular function in small animals.

UNLABELLED: 18F-FDG PET can identify areas of myocardial viability and necrosis and provide useful information on the effectiveness of experimental techniques designed to improve contractile function and myocardial vascularization in small animals. The left ventricular volume (LVV) and left ventricular ejection fraction (LVEF) in normal and diseased rats were measured in vivo using the high-resolution avalanche photodiode (APD) small-animal PET scanner of the Université de Sherbrooke. The measurements obtained by PET were compared with those obtained by high-resolution echocardiography and with known values obtained from a small, variable-volume cardiac phantom. METHODS: List-mode gated (18)F-FDG PET studies were performed using the APD PET scanner on 30 rats: 11 healthy, 4 under septic shock, and 15 with heart failure induced by ligature of the left coronary artery. PET images were resized to match human-scale pixels and analyzed using a standard clinical cardiac software program. The LVV and LVEF from the same animals were also evaluated by echocardiography. RESULTS: Agreement was excellent between the endocardial volumes determined by PET and the actual volumes of the cardiac phantom (r(2) = 0.96). Agreement between PET and echocardiography for LVV ranged from good in healthy rats (r(2) = 0.89) to fair in diseased rats (r(2) = 0.49). Agreement was fair between LVEF values measured by the 2 methods (r(2) = 0.56). Normal rats had an average LVEF of 83.2% +/- 8.0% using PET and 81.6% +/- 6.0% using echocardiography. In rats with heart failure, LVEF was 54.6% +/- 15.9% using PET and 54.2% +/- 13.3% using echocardiography. CONCLUSION: Both PET and echocardiography clearly differentiated normal rats from rats with heart failure. Echocardiography is fast and convenient, whereas list-mode PET is also able to assess defect size, myocardial viability, and metabolism.