Laurent Pinot

A new high resolution radioimager for the quantitative analysis of radiolabelled molecules in tissue section

We present a high-speed, high-resolution imager of beta particles. It is devoted to be used in autoradiography experiments such as receptor binding or in situ hybridization experiments, either instead of, or in complement with autoradiographic film and emulsions. It allows the user to locate and perform quantitative analyses of (3H, 14C, 35S, 33P, 32P, 125I) labelled molecules with a 15 microm spatial resolution on a 0.9 x 1.3 cm2 sensitive area. Combining recent techniques (specific scintillator thin sheets and intensified charge-coupled device (CCD)) this imager offers a wide dynamic range and real-time acquisition.

In vivo quantification of localized neuronal activation and inhibition in the rat brain using a dedicated high temporal-resolution β+-sensitive microprobe

Understanding brain disorders, the neural processes implicated in cognitive functions and their alterations in neurodegenerative pathologies, or testing new therapies for these diseases would benefit greatly from combined use of an increasing number of rodent models and neuroimaging methods specifically adapted to the rodent brain. Besides magnetic resonance (MR) imaging and functional MR, positron-emission tomography (PET) remains a unique methodology to study in vivo brain processes. However, current high spatial-resolution tomographs suffer from several technical limitations such as high cost, low sensitivity, and the need of restraining the animal during image acquisition. We have developed a β+-sensitive high temporal-resolution system that overcomes these problems and allows the in vivo quantification of cerebral biochemical processes in rodents. This β-MICROPROBE is an in situ technique involving the insertion of a fine probe into brain tissue in a way very similar to that used for microdialysis and cell electrode recordings. In this respect, it provides information on molecular interactions and pathways, which is complementary to that produced by these technologies as well as other modalities such as MR or fluorescence imaging. This study describes two experiments that provide a proof of concept to substantiate the potential of this technique and demonstrate the feasibility of quantifying brain activation or metabolic depression in individual living rats with 2-[18F]fluoro-2-deoxy-d-glucose and standard compartmental modeling techniques. Furthermore, it was possible to identify correctly the origin of variations in glucose consumption at the hexokinase level, which demonstrate the strength of the method and its adequacy for in vivo quantitative metabolic studies in small animals.

Physical Performance of an Intraoperative Beta Probe Dedicated to Glioma Radioguided Surgery

The precise delineation and excision of brain tumor extent allows to improve survival outcome and quality of life of surgically treated patients. In order to refine the resection of gliomas, we are developing a novel intraoperative probe specifically dedicated to the localization of residual tumor after the bulk has been excised. The probe, built around clear and plastic scintillating fibers, was designed to detect positrons emitted from radiolabeled brain tissue in order to discriminate more specifically neoplastic from normal tissues. The probe was also built to be directly coupled to the excision tool leading to simultaneous detection and removal of tumor. We report here performances of the first radio-isotopic configuration of the intraoperative probe which consists of a detection head composed of eight detection elements held around the excision tool in a closed packed annular arrangement. This head is coupled to an optic fiber bundle that exports the scintillating light to a multi-channel photomultiplier tube. The gamma ray background generated by the annihilation of beta+ in tissues is eliminated by a real-time subtraction method. The detector exhibits a beta sensitivity of 139 cps/kBq and a gamma ray rejection efficiency of 99.5%. The ability of the probe to detect residual lesions was evaluated with a realistic brain phantom representing the surgical cavity and the boundaries of the tumor. We showed that lesions as small as 5 mm in diameter can be detected for tumor to normal tissue uptake ratios of fluorinated tracers greater than 3.5. This ratio is achieved with radiopharmaceuticals like 18F-FET or 18F-choline. These promising results suggest that the features of our system are compatible with in situ localization of residual radiolabeled tumors.

Simultaneous in vivo magnetic resonance imaging and radioactive measurements with the β-MicroProbe

PurposeMultimodal instrumentation is a new technical approach allowing simultaneous and complementary in vivo recordings of complementary biological parameters. To elucidate further the physiopathological mechanisms in intact small animal models, especially for brain studies, a challenging issue is the actual coupling of magnetic resonance imaging (MRI) techniques with positron emission tomography (PET): it has been shown that running the technology for radioactive imaging in a magnet alters the spatiotemporal performance of both modalities. Thus, we propose an alternative coupling of techniques that uses the β-MicroProbe instead of PET for local measurements of radioactivity coupled with MRI.MethodsWe simultaneously recorded local radioactivity due to [18F]MPPF (a 5-HT1A receptor PET radiotracer) binding in the hippocampus with the β-MicroProbe and carried out anatomical MRI in the same anaesthetised rat.ResultsThe comparison of [18F]MPPF kinetics obtained from animals in a magnet with kinetics from a control group outside the magnet allowed us to determine the stability of tracer biokinetic measurements over time in the magnet. We were thus able to show that the β-MicroProbe reliably measures radioactivity in rat brains under an intense magnetic field of 7 Tesla.ConclusionThe biological validation of a β-MicroProbe/MRI dual system reported here opens up a wide range of future multimodal approaches for functional and pharmacological measurements by the probe combined with various magnetic resonance technologies, including anatomical MRI, functional MRI and MR spectroscopy.

Combining the radiosensitive Beta MicroProbe to Nuclear Magnetic Resonance: theoretical approach for in vivo studies in small animals

In vivo small animal imaging with multiple modalities has become an important tool in modern biomedical research. Indeed, combining exploratory techniques allows simultaneous recording of complementary data, which is required to elucidate complex physiopathological mechanisms. In this field, because of strict technical constraints in vivo, an exciting challenge remains in the combination of Nuclear Magnetic Resonance (NMR) and Positron Emission Tomography (PET). Coupling NMR with a radiosensitive Beta MicroProbe offers therefore a very interesting technical alternative. Here, we assessed the feasibility of this new combination by theoretically evaluating the ability of the Beta MicroProbe to monitor radioactivity in a magnet. To that aim, we modelled with Geant4 the effect of an intense magnetic field on the probe field of view and showed that the field should not have an impact on the global efficiency of the probe.

The Potential of a Radiosensitive Intracerebral Probe to Monitor 18F-MPPF Binding in Mouse Hippocampus In Vivo

As mouse imaging has become more challenging in preclinical research, efforts have been made to develop dedicated PET systems. Although these systems are currently used for the study of physiopathologic murine models, they present some drawbacks for brain studies, including a low temporal resolution that limits the pharmacokinetic study of radiotracers. The aim of this study was to demonstrate the ability of a radiosensitive intracerebral probe to measure the binding of a radiotracer in the mouse brain in vivo. Methods: The potential of a probe 0.25 mm in diameter for pharmacokinetic studies was assessed. First, Monte Carlo simulations followed by experimental studies were used to evaluate the detection volume and sensitivity of the probe and its adequacy for the size of loci in the mouse brain. Second, ex vivo autoradiography of 5-hydroxytryptamine receptor 1A (5-HT1A) receptors in the mouse brain was performed with the PET radiotracer 2′-methoxyphenyl-(N-2′-pyridinyl)-p-18F-fluorobenzamidoethylpiperazine (18F-MPPF). Finally, the binding kinetics of 18F-MPPF were measured in vivo in both the hippocampus and the cerebellum of mice. Results: Both the simulations and the experimental studies demonstrated the feasibility of using small probes to measure radioactive concentrations in specific regions of the mouse brain. Ex vivo autoradiography showed a heterogeneous distribution of 18F-MPPF consistent with the known distribution of 5-HT1A in the mouse brain. Finally, the time–activity curves obtained in vivo were reproducible and validated the capacity of the new probe to accurately measure 18F-MPPF kinetics in the mouse hippocampus. Conclusion: Our results demonstrate the ability of the tested radiosensitive intracerebral probe to monitor binding of PET radiotracers in anesthetized mice in vivo, with high temporal resolution suited for compartmental modeling.