Principles of nuclear medicine imaging: planar, SPECT, PET, multi-modality, and autoradiography systems

The underlying principles of nuclear medicine imaging involve the use of unsealed sources of radioactivity in the form of radiopharmaceuticals. The ionizing radiations that accompany the decay of the administered radioactivity can be quantitatively detected, measured, and imaged in vivo with instruments such as gamma cameras. This paper reviews the design and operating principles, as well as the capabilities and limitations, of instruments used clinically and preclinically for in vivo radionuclide imaging. These include gamma cameras, single-photon emission computed tomography (SPECT) scanners, and positron emission tomography (PET) scanners. The technical basis of autoradiography is reviewed as well.     

Advanced radionuclide detection techniques for in vitro and in vivo animal imaging.

This review of the different methodologies used for animal imaging with radioactive compounds presents the most recent approaches developed for both in vitro and in vivo studies. The choice of a detector for analysis of the spatial distribution of radionuclides deposited in biological tissues results in a trade-off between the size and nature of the region to study (in vitro or in vivo), the required spatial resolution and the penetrating characteristics of the ionizing radiation. Real time detectors are now available for quantitative imaging of 2D or 3D radioactive samples and offer either an increased dynamic range or a lowered sensitivity in comparison with film radioautography. For high resolution imaging, two specific techniques are proposed for applications to rodents. The usefulness of self-triggering intensified charge coupled device (STIC) is illustrated for in vitro localization in radiotoxicological studies of alpha-emitters. For in vivo techniques, the performance of positron emission tomography (PET) is discussed, as a promising method of molecular imaging of biological processes.     

Cerenkov Luminescence Imaging (CLI) for Cancer Therapy Monitoring

In molecular imaging, positron emission tomography (PET) and optical imaging (OI) are two of the most important and thus most widely used modalities^1-3. PET is characterized by its excellent sensitivity and quantification ability while OI is notable for non-radiation, relative low cost, short scanning time, high throughput, and wide availability to basic researchers. However, both modalities have their shortcomings as well. PET suffers from poor spatial resolution and high cost, while OI is mostly limited to preclinical applications because of its limited tissue penetration along with prominent scattering optical signals through the thickness of living tissues.Recently a bridge between PET and OI has emerged with the discovery of Cerenkov Luminescence Imaging (CLI)^4-6. CLI is a new imaging modality that harnesses Cerenkov Radiation (CR) to image radionuclides with OI instruments. Russian Nobel laureate Alekseyevich Cerenkov and his colleagues originally discovered CR in 1934. It is a form of electromagnetic radiation emitted when a charged particle travels at a superluminal speed in a dielectric medium^7,8. The charged particle, whether positron or electron, perturbs the electromagnetic field of the medium by displacing the electrons in its atoms. After passing of the disruption photons are emitted as the displaced electrons return to the ground state. For instance, one ¹⁸F decay was estimated to produce an average of 3 photons in water⁵. Since its emergence, CLI has been investigated for its use in a variety of preclinical applications including in vivo tumor imaging, reporter gene imaging, radiotracer development, multimodality imaging, among others^4,5,9,10,11. The most important reason why CLI has enjoyed much success so far is that this new technology takes advantage of the low cost and wide availability of OI to image radionuclides, which used to be imaged only by more expensive and less available nuclear imaging modalities such as PET.Here, we present the method of using CLI to monitor cancer drug therapy. Our group has recently investigated this new application and validated its feasibility by a proof-of-concept study¹². We demonstrated that CLI and PET exhibited excellent correlations across different tumor xenografts and imaging probes. This is consistent with the overarching principle of CR that CLI essentially visualizes the same radionuclides as PET. We selected Bevacizumab (Avastin; Genentech/Roche) as our therapeutic agent because it is a well-known angiogenesis inhibitor^13,14. Maturation of this technology in the near future can be envisioned to have a significant impact on preclinical drug development, screening, as well as therapy monitoring of patients receiving treatments.     

Antibody fragments labeled with fluorine-18 and gallium-68: In vivo comparison with indium-111 and iodine-125-labeled fragments

Although monoclonal antibodies have been radiolabeled with many different radionuclides, the application of positron emission tomography (PET) to the imaging of radiolabeled antibodies has been limited to the investigation of a small number of long-lived radionuclides. In this study, we labeled F(ab′)₂ fragments of a mouse monoclonal antibody (BB5-G1) specific for a human parathyroid surface antigen with the positron emitting radionuclides, gallium-68 and fluorine-18. The biodistribution of the fragments was evaluated in a nude mice model and the results were compared to those obtained with fragments labeled with iodine-125 and indium-111 using conventional labeling techniques. All labeled fragments bound to human parathyroid tissue implanted in nude mice, with parathyroid-to-muscle ratios reaching as high as 10:1, 4 h after administration. A major difference was observed in the uptake and clearance of the various labeled fragments through the kidney. The halogen activity cleared, but the metal radioactivity was retained in the kidney. The results indicate that the fluorine-18 or gallium-68 labeled fragment may be useful for parathyroid imaging with positron emission tomography.     

Targeting peptides and positron emission tomography

Biologically active peptides have during the last decades made their way into conventional nuclear medicine diagnosis using single photon emission computed tomography (SPECT) and gamma-camera. Several clinical trails are also investigating the role of radiolabeled peptides for targeting radionuclide therapy. This has raised the question as to whether positron emission tomography (PET) can be used in order to obtain better quantitative information of the peptide distribution in tumor and healthy organs, i.e., to get a better dosimetry. Positron emitting analogs of the therapeutic radionuclides used have been produced and successfully applied in peptide pharmacokinetic measurements with PET. But the recent boom in (18)FDG-PET ((18)FDG = [(18)F]2-deoxy-2-fluoro-D-glucose), and with this a worldwide increasing number of PET systems, has also inspired several research groups to hunt for alternative labels to be used for peptide diagnostics and PET. The rapid kinetic of short peptides agrees well with the short half-lives of standard PET nuclides like (11)C and (18)F. Especially, (18)F appears to be excellent for labeling bioactive peptides due to its favorable physical and nuclear characteristics. However, with present techniques labeling peptides with (18)F is laborious and time-consuming, and is not yet a clinical alternative. Other halogens like (75, 76)Br and (124)I are, from the chemical point of view, easier to apply. But an even better labeling alternative may be positron emitting metal ions like (55)Co, (68)Ga, and (110m)In since they tend to give better intracellular retention and thus a better signal-to-background ratio than the halogen labels. The main drawback with these radionuclides is that they are not readily available. Some of these radionuclides also emit gamma in their decay that may affect the measuring properties of the PET equipment. This article reviews mainly the present situation of production and use of nonconventional positron emitters for peptide labeling.     

PET as a potential tool for imaging molecular mechanisms of oncology in man

During the past ten years, positron emission tomography (PET) has been increasingly developed for imaging and quantifying molecular mechanisms in oncology. The technique uses radionuclides to label molecules, which can then be imaged in man. The inherent sensitivity and specificity of PET is unrivalled because it can image molecular interactions and pathways, providing quantitative kinetic information down to the subpicomolar level. This technology has the potential to answer a large number of important clinical questions in translational research in oncology. However, the challenges in the methodology are substantial. Molecular imaging has the potential to assist in the optimization of molecular-based targeted therapies in cancer and to investigate the function of the genome.     

The Bioconjugation and Radiosynthesis of 89Zr-DFO-labeled Antibodies

The exceptional affinity, specificity, and selectivity of antibodies make them extraordinarily attractive vectors for tumor-targeted PET radiopharmaceuticals. Due to their multi-day biological half-life, antibodies must be labeled with positron-emitting radionuclides with relatively long physical decay half-lives. Traditionally, the positron-emitting isotopes ¹²⁴I (t_1/2 = 4.18 d), ⁸⁶Y (t_1/2 = 14.7 hr), and ⁶⁴Cu (t_1/2 = 12.7 hr) have been used to label antibodies for PET imaging. More recently, however, the field has witnessed a dramatic increase in the use of the positron-emitting radiometal ⁸⁹Zr in antibody-based PET imaging agents. ⁸⁹Zr is a nearly ideal radioisotope for PET imaging with immunoconjugates, as it possesses a physical half-life (t_1/2 = 78.4 hr) that is compatible with the in vivo pharmacokinetics of antibodies and emits a relatively low energy positron that produces high resolution images. Furthermore, antibodies can be straightforwardly labeled with ⁸⁹Zr using the siderophore-derived chelator desferrioxamine (DFO). In this protocol, the prostate-specific membrane antigen targeting antibody J591 will be used as a model system to illustrate (1) the bioconjugation of the bifunctional chelator DFO-isothiocyanate to an antibody, (2) the radiosynthesis and purification of a ⁸⁹Zr-DFO-mAb radioimmunoconjugate, and (3) in vivo PET imaging with an ⁸⁹Zr-DFO-mAb radioimmunoconjugate in a murine model of cancer.     

Measuring drug-related receptor occupancy with positron emission tomography

Several techniques can be used to measure indirectly the effect of drugs (e.g., EEG, fMRI) in healthy volunteers and in patients. Although each technique has its merits, a direct link between drug efficacy and site of action in vivo usually cannot be established. In addition, when the specific mode of action of a drug has been determined from preclinical studies, it is often not known whether the administered dose is optimal for humans. Both industry and academia are becoming more and more interested in determining the dose-related occupancy of specific targets caused by administration of drugs under test. Positron emission tomography (PET) and single-photon emission computed tomography (SPECT) are noninvasive imaging techniques that can give insight into the relationship between target occupancy and drug efficacy, provided a suitable radioligand is available. Although SPECT has certain advantages (e.g., a long half-life of the radionuclides), the spatial and temporal resolution as well as the labeling possibilities of this technique are limited. This review focuses on PET methodology for conducting drug occupancy studies in humans.     

ClearPEM,un TEP dédié au sein

Conventional breast imaging (mammography, ultrasound, MRI) relies on the analysis of anatomical characteristics. Whole-body positron emission tomography (PET) allows for the acquisition of a metabolic image, yet is limited by its poor spatial resolution. The Crystal Clear Collaboration developed a PET dedicated for breast imaging, the ClearPEM, in order to offer a high-resolution nuclear imaging technique. The patient is installed in the prone position on the exam bed, with two detector plates rotating around to breast to acquire a 3-dimensional image. Two prototypes were built and installed in hospitals. We summarize the technical solutions necessary for the development of this system and present a summary of its performances as well as an outlook on preclinical and clinical tests.     

18F]FDG labeling of neural stem cells for in vivo cell tracking with positron emission tomography: inhibition of tracer release by phloretin

The introduction of neural stem cells into the brain has promising therapeutic potential for the treatment of neurodegenerative diseases. To monitor the cellular replacement therapy, that is, to determine stem cell migration, survival, and differentiation, in vivo tracking methods are needed. Ideally, these tracking methods are noninvasive. Noninvasive tracking methods that have been successfully used for the visualization of blood-derived progenitor cells include magnetic resonance imaging and radionuclide imaging using single-photon emission computed tomography (SPECT) and positron emission tomography (PET). The SPECT tracer In-111-oxine is suitable for stem cell labeling, but for studies in small animals, the higher sensitivity and facile quantification that can be obtained with PET are preferred. Here the potential of 2'-[18F]fluoro-2'-deoxy-D-glucose ([18F]-FDG), a PET tracer, for tracking of neural stem cell (NSCs) trafficking toward an inflammation site was investigated. [18F]-FDG turns out to be a poor radiopharmaceutical to label NSCs owing to the low labeling efficiency and substantial release of radioactivity from these cells. Efflux of [18F]-FDG from NSCs can be effectively reduced by phloretin in vitro, but inhibition of tracer release is insufficient in vivo for accurate monitoring of stem cell trafficking.     

A versatile bifunctional chelate for radiolabeling humanized anti-CEA antibody with In-111 and Cu-64 at either thiol or amino groups: PET imaging of CEA-positive tumors with whole antibodies

Radiolabeled anti-carcinoembryonic antigen (CEA) antibodies have the potential to give excellent images of a wide variety of human tumors, including tumors of the colon, breast, lung, and medullar thyroid. In order to realize the goals of routine and repetitive clinical imaging with anti-CEA antibodies, it is necessary that the antibodies have a high affinity for CEA, low cross reactivity and uptake in normal tissues, and low immunogenicity. The humanized anti-CEA antibody hT84.66-M5A (M5A) fulfills these criteria with an affinity constant of >10 (10) M (-1), no reactivity with CEA cross-reacting antigens found in normal tissues, and >90% human protein sequence. A further requirement for routine clinical use of radiolabeled antibodies is a versatile method of radiolabeling that allows the use of multiple radionuclides that differ in their radioemissions and half-lives. We describe a versatile bifunctional chelator, DO3A-VS (1,4,7-tris(carboxymethyl)-10-(vinylsulfone)-1,4,7,10-tetraazacyclododecane) that binds a range of radiometals including 111 In for gamma-ray imaging and 64Cu for positron emission tomography (PET), and which can be conjugated with negligible loss of immunoreactivity either to sulfhydryls (SH) in the hinge region of lightly reduced immunoglobulins or surface lysines (NH) of immunoglobulins. Based on our correlative studies comparing the kinetics of radiolabeled anti-CEA antibodies in murine models with those in man, we predict that 64Cu-labeled intact, humanized antibodies can be used to image CEA positive tumors in the clinic.     

Current and future anatomical and functional imaging approaches to pheochromocytoma and paraganglioma

After establishing a biochemical diagnosis, pheochromocytomas and extra-adrenal paragangliomas (PPGLs) can be localized using different anatomical and functional imaging modalities. These include computed tomography, magnetic resonance imaging, single-photon emission computed tomography (SPECT) using 123I-metaiodobenzylguanidine or 111In-DTPA-pentetreotide, and positron emission tomography (PET) using 6-[18F]-fluorodopamine (18F-FDA), 6-[18F]-fluoro-l-3,4-dihydroxyphenylalanine (18F-DOPA), and 2-[18F]-fluoro-2-deoxy-d-glucose. We review the currently available data on the performance of anatomical imaging, SPECT, and PET for the detection of (metastatic) PPGL as well as parasympathetic head and neck paragangliomas. We show that there appears to be no 'gold-standard' imaging technique for all patients with (suspected) PPGL. A tailor-made approach is warranted, guided by clinical, biochemical, and genetic characteristics. In the current era of a growing number of PET tracers, PPGL imaging has moved beyond tumor localization towards functional characterization of tumors.     

Light sharing in multi-flat-panel-PMT PEM detectors

Large are a detectors, such as those used in positron emission mammography (PEM) and scintimammography, utilize arrays of discrete semtillator elements mounted on arrays of position sensitive photomultiplier tubes (PSPMT). Scintillator elements can be packed very densely (minimizing area between elements), allowing good detection sensitivity and spatial resolution. And, while new flat panel PSPMTS have minimal inactive edges, when they are placed in arrays significant dead spaces where scintillation light is undetectable are created. To address this problem, a light guide is often placed between the detector and PSPMT array to spread scintillation light so that these gaps can be bridged. In this investigation we studied the effect of light guides of various thickness on system performance. A 10×10 element array of LYSO detector elements was coupled to the center of a 2×2 array of PSPMTs through varying thicknesses (1 to 4 mm) of UV glass. The spot size of the imaged elements and distortions in the regular square pattern of the imaged scintillator arrays were evaluated. Energy resolution was measured by placing single elements of LYSO at several locations of the PSPMT array. Spatial distortions in the images of the array were reduced by using thicker light guides (3–4 mm). Use of thicker light guides, however, resulted in reduced pixel resolution and slight degradation of energy resolution. Therefore, some loss of pixel and energy resolution will accompany the use of thick light guides (minimum of 3 mm) required for optimum identification of detector elements.     

Real time non-invasive imaging of receptor-ligand interactions in vivo

Non-invasive longitudinal detection and evaluation of gene expression in living animals can provide investigators with an understanding of the ontogeny of a gene's biological function(s). Currently, mouse model systems are used to optimize magnetic resonance imaging (MRI), positron emission tomography (PET), single-photon emission computed tomography (SPECT), and optical imaging modalities to detect gene expression and protein function. These molecular imaging strategies are being developed to assess tumor growth and the tumor microenvironment. In addition, pre-labeling of progenitor cells can provide invaluable information about the developmental lineage of stem cells both in organogenesis and tumorigenesis. The feasibility of this approach has been extensively tested by targeting of endogenous tumor cell receptors with labeled ligand (or ligand analog) reporters and targeting enzymes with labeled substrate (or substrate analog). We will primarily discuss MRI, PET, and SPECT imaging of cell surface receptors and the feasibility of non-invasive imaging of gene expression using the tumor microenvironment (e.g., hypoxia) as a conditional regulator of gene expression.     

Assessment of cardiovascular apoptosis in the isolated rat heart by magnetic resonance molecular imaging

Apoptosis, an active process of cell self-destruction, is associated with myocardial ischemia. The redistribution of phosphatidylserine (PS) from the inner to the outer leaflet of the cell membrane is an early event in apoptosis. Annexin V, a protein with high specificity and tight binding to PS, was used to identify and localize apoptosis in the ischemic heart.Fluorescein-labeled annexin V has been used routinely for the assessment of apoptosis in vitro. For the detection of apoptosis in vivo, positron emission tomography and single-photon emission computed tomography have been shown to be suitable tools. In view of the relatively low spatial resolution of nuclear imaging techniques, we developed a high-resolution contrast-enhanced magnetic resonance imaging (MRI) method that allows rapid and noninvasive monitoring of apoptosis in intact organs. Instead of employing superparamagnetic iron oxide particles linked to annexin V, a new T1 contrast agent was used. To this effect, annexin V was linked to gadolinium diethylenetriamine pentaacetate (Gd-DTPA)-coated liposomes.The left coronary artery of perfused isolated rat hearts was ligated for 30 min followed by reperfusion. T(1) and T(2)* images were acquired by using an 11.7-T magnet before and after intracoronary injection of Gd-DTP-labeled annexin V to visualize apoptotic cells. A significant increase in signal intensity was visible in those regions containing cardiomyocytes in the early stage of apoptosis. Because labeling of early apoptotic cell death in intact organs by histological and immunohistochemical methods remains challenging, the use of Gd-DTPA-labeled annexin V in MRI is clearly an improvement in rapid targeting of apoptotic cells in the ischemic and reperfused myocardium.     

Radiolabelled proteins for positron emission tomography: Pros and cons of labelling methods

Background

Dynamic biomedical research is currently yielding a wealth of information about disease-associated molecular alterations on cell surfaces and in the extracellular space. The ability to visualize and quantify these alterations in vivo could provide important diagnostic information and be used to guide individually-optimized therapy. Biotechnology can provide proteinaceous molecular probes with highly specific target recognitions. Suitably labelled, these may be used as tracers for radionuclide-based imaging of molecular disease signatures. If the labels are positron-emitting radionuclides, the superior resolution, sensitivity and quantification capability of positron emission tomography (PET) can be exploited.

Scope of review

This article discusses different approaches to labelling proteins with positron-emitting nuclides with suggestions made depending on the biological features of the tracers.

Major conclusions

Factors such as matching biological and physical half-lives, availability of the nuclide, labelling yields, and influences of labelling on targeting properties (affinity, charge and lipophilicity, cellular processing and retention of catabolites) should be considered when selecting a labelling strategy for each proteinaceous tracer.

General significance

The labelling strategy used can make all the difference between success and failure in a tracer application. This review emphasises chemical, biological and pharmacological considerations in labelling proteins with positron-emitting radionuclides.     

CZT gamma camera for scintimammography

A high performance prototype gamma camera based on the semiconductor radiation detector Cd(Zn)Te is described. The camera features high spatial resolution, high-energy resolution, a reduced dead space on the edge of the field of view, and a compact format. The camera performance was first examined by comparison of small field of view examinations with those from an Elscint SP6HR standard clinical gamma camera. The new camera was found to give equal or improved image quality. The camera was then used for a systematic phantom study of small lesions in a background as would be found in breast cancer imaging. In this study the camera was able to systematically detect smaller, deeper, and fainter lesions. The camera is presently being used in a clinical trial aimed to assess its value in scintimammography where previous limitations of image quality and detector size have restricted the use of the functional imaging techniques. Preliminary results from 40 patients show high sensitivity and specificity with respect to X-ray mammography and surgery.     

Therapy region monitoring based on PET using 478 keV single prompt gamma ray during BNCT: A Monte Carlo simulation study

We confirmed the feasibility of using our proposed system to extract two different kinds of functional images from a positron emission tomography (PET) module by using an insertable collimator during boron neutron capture therapy (BNCT). Coincidence events from a tumor region that included boron particles were identified by a PET scanner before BNCT; subsequently, the prompt gamma ray events from the same tumor region were collected after exposure to an external neutron beam through an insertable collimator on the PET detector. Five tumor regions that contained boron particles and were located in the water phantom and in the BNCT system with the PET module were simulated with Monte Carlo simulation code. The acquired images were quantitatively analyzed. Based on the receiver operating characteristic (ROC) curves in the five boron regions, A, B, C, D, and E, the PET and single-photon images were 10.2%, 11.7%, 8.2% (center region), 12.6%, and 10.5%, respectively. We were able to acquire simultaneously PET and single prompt photon images for tumor regions monitoring by using an insertable collimator without any additional isotopes.