Cerenkov luminescence and PET imaging of 90Y: capabilities and limitations in small animal applications.

The in vivo sensitivity limits and quantification performance of Cerenkov luminescence imaging have been studied using a tissue-like mouse phantom and 90Y. For a small, 9 mm deep target in the phantom, with no background activity present, the Cerenkov luminescence 90Y detection limit determined from contrast-to-noise ratios is 10 nCi for a 2 min exposure with a sensitive CCD camera and no filters. For quantitative performance, the values extracted from regions of interest on the images are linear within 5% of a straight line fit versus target activity for target activity of 70 nCi and above. The small branching ratio to decay with positron emission for 90Y also permits low-statistics PET imaging of the radionuclide. For PET imaging of the same phantom, with a small animal LSO detector-based scanner, the 90Y detection limit is approximately 3 orders of magnitude higher at 10 µCi.

Imaging Salt Uptake Dynamics in Plants Using PET.

Soil salinity is a global environmental challenge for crop production. Understanding the uptake and transport properties of salt in plants is crucial to evaluate their potential for growth in high salinity soils and as a basis for engineering varieties with increased salt tolerance. Positron emission tomography (PET), traditionally used in medical and animal imaging applications for assessing and quantifying the dynamic bio-distribution of molecular species, has the potential to provide useful measurements of salt transport dynamics in an intact plant. Here we report on the feasibility of studying the dynamic transport of 22Na in millet using PET. Twenty-four green foxtail (Setaria viridis L. Beauv.) plants, 12 of each of two different accessions, were incubated in a growth solution containing 22Na+ ions and imaged at 5 time points over a 2-week period using a high-resolution small animal PET scanner. The reconstructed PET images showed clear evidence of sodium transport throughout the whole plant over time. Quantitative region-of-interest analysis of the PET data confirmed a strong correlation between total 22Na activity in the plants and time. Our results showed consistent salt transport dynamics within plants of the same variety and important differences between the accessions. These differences were corroborated by independent measurement of Na+ content and expression of the NHX transcript, a gene implicated in sodium transport. Our results demonstrate that PET can be used to quantitatively evaluate the transport of sodium in plants over time and, potentially, to discern differing salt-tolerance properties between plant varieties. In this paper, we also address the practical radiation safety aspects of working with 22Na in the context of plant imaging and describe a robust pipeline for handling and incubating plants. We conclude that PET is a promising and practical candidate technology to complement more traditional salt analysis methods and provide insights into systems-level salt transport mechanisms in intact plants.

Real-time whole-plant dynamics of heavy metal transport in Arabidopsis halleri and Arabidopsis thaliana by gamma-ray imaging.

Heavy metals such as zinc are essential for plant growth, but toxic at high concentrations. Despite our knowledge of the molecular mechanisms of heavy metal uptake by plants, experimentally addressing the real-time whole-plant dynamics of heavy metal uptake and partitioning has remained a challenge. To overcome this, we applied a high sensitivity gamma-ray imaging system to image uptake and transport of radioactive 65Zn in whole-plant assays of Arabidopsis thaliana and the Zn hyperaccumulator Arabidopsis halleri. We show that our system can be used to quantitatively image and measure uptake and root-to-shoot translocation dynamics of zinc in real time. In the metal hyperaccumulator Arabidopsis halleri, 65Zn uptake and transport from its growth media to the shoot occurs rapidly and on time scales similar to those reported in rice. In transgenic A. halleri plants in which expression of the zinc transporter gene HMA4 is suppressed by RNAi, 65Zn uptake is completely abolished.

First Cerenkov charge-induction (CCI) TlBr detector for TOF-PET and proton range verification.

Thallium bromide (TlBr) is a semiconductor material and, simultaneously, a good Cerenkov radiator. The performance of a TlBr detector that integrates two different readouts, the charge induction readout and the detection of Cerenkov light, was evaluated. A TlBr detector with dimensions of 4 × 4 × 5 mm3, with a monolithic cathode and an anode segmented into strips, was manufactured. One of the bare and polished 4 × 4 mm2 faces of the detector was coupled to a silicon photomultiplier (SiPM) to read out the Cerenkov light. Simultaneous timing and energy resolutions of <400 ps full width at half maximum (FWHM) and ~8.5% at 511 keV were measured using the Cerenkov detection and charge induction readouts, respectively. A coincidence time resolution of 330 ps was obtained when selecting Cerenkov events with amplitudes above 70 mV. The combination of both readouts showed the potential to resolve the depth-of-interaction (DOI) positioning, based on the improvement of energy resolution when selecting events with similar electron drift times. This manuscript sets the stage for a new family of semiconductor detectors that combine charge induction readout with the Cerenkov light detection. Such detectors can provide, simultaneously, outstanding timing, energy, and spatial resolution, and will be an excellent fit for applications that require the detection of high-energy gamma photons with high timing accuracy, such as time-of-flight positron emission tomography (TOF-PET) and prompt gamma imaging (PGI) to assess the particle range in hadron therapy.

Theoretical investigation of ultrasound-modulated Cerenkov luminescence imaging for higher-resolution imaging in turbid media.

Cerenkov luminescence imaging (CLI) is an optical technique for imaging radiolabeled molecules in vivo. It has demonstrated utility in both the clinical and preclinical settings and can serve as a substitute for nuclear imaging instrumentation in some cases. However, optical scattering fundamentally limits the resolution and depth of imaging that can be achieved with this modality. In this Letter, we report the numerical results that support the potential for ultrasound-modulated Cerenkov luminescence imaging (USCLI), a new imaging modality that can mitigate optical scattering. The technique uses an acoustic field to modulate the refractive index of the medium and, thus, the intensity of Cerenkov luminescence in a spatially precise manner. This mechanism of contrast has not been reported previously. For a physiologically compatible ultrasound peak pressure of 1 MPa, ∼0.1% of the Cerenkov signal can be modulated. Furthermore, our simulations show that USCLI can overcome the scattering limit of resolution for CLI and provide higher-resolution imaging. For an F18 point source centered in a 1 cm3 simulated tissue phantom with a scattering coefficient of µs'=10 cm-1, <2 mm full width at half-maximum lateral spatial resolution is possible, a resolution three times finer than the same phantom imaged with CLI.

Development of TlBr detectors for PET imaging.

Thallium bromide (TlBr) is a promising semiconductor detector material for positron emission tomography (PET) because it can offer very good energy resolution and 3D segmentation capabilities, and it also provides detection efficiency surpassing that of commonly used scintillators. Energy, timing, and spatial resolution were measured for thin (<1 mm) TlBr detectors. The energy and timing resolution were measured simultaneously for the same planar 0.87 mm-thick TlBr device. An energy resolution of (6.4 ± 1.3)% at 511 keV was achieved at -400 V bias voltage and at room temperature. A timing resolution of (27.8 ± 4.1) ns FWHM was achieved for the same operating conditions when appropriate energy gating was applied. The intrinsic spatial resolution was measured to be 0.9 mm FWHM for a TlBr detector with metallic strip contacts of 0.5 mm pitch. As material properties improve, higher bias voltage should improve timing performance. A stack of thin detectors with finely segmented readout can create a modular detector with excellent energy and spatial resolution for PET applications.

Towards time-of-flight PET with a semiconductor detector.

The feasibility of using Cerenkov light, generated by energetic electrons following 511 keV photon interactions in the semiconductor TlBr, to obtain fast timing information for positron emission tomography (PET) was evaluated. Due to its high refractive index, TlBr is a relatively good Cerenkov radiator and with its wide bandgap, has good optical transparency across most of the visible spectrum. Coupling an SiPM photodetector to a slab of TlBr (TlBr-SiPM) yielded a coincidence timing resolution of 620 ps FWHM between the TlBr-SiPM detector and a LFS reference detector. This value improved to 430 ps FWHM by applying a high pulse amplitude cut based on the TlBr-SiPM and reference detector signal amplitudes. These results are the best ever achieved with a semiconductor PET detector and already approach the performance required for time-of-flight. As TlBr has higher stopping power and better energy resolution than the conventional scintillation detectors currently used in PET scanners, a hybrid TlBr-SiPM detector with fast timing capability becomes an interesting option for further development.

Quantitative assessment of Cerenkov luminescence for radioguided brain tumor resection surgery.

Cerenkov luminescence imaging (CLI) is a developing imaging modality that detects radiolabeled molecules via visible light emitted during the radioactive decay process. We used a Monte Carlo based computer simulation to quantitatively investigate CLI compared to direct detection of the ionizing radiation itself as an intraoperative imaging tool for assessment of brain tumor margins. Our brain tumor model consisted of a 1 mm spherical tumor remnant embedded up to 5 mm in depth below the surface of normal brain tissue. Tumor to background contrast ranging from 2:1 to 10:1 were considered. We quantified all decay signals (e±, gamma photon, Cerenkov photons) reaching the brain volume surface. CLI proved to be the most sensitive method for detecting the tumor volume in both imaging and non-imaging strategies as assessed by contrast-to-noise ratio and by receiver operating characteristic output of a channelized Hotelling observer.

Computed Cerenkov luminescence yields for radionuclides used in biology and medicine.

Cerenkov luminescence imaging is an emerging biomedical imaging modality that takes advantage of the optical Cerenkov photons emitted following the decay of radionuclides in dielectric media such as tissue. Cerenkov radiation potentially allows many biomedically-relevant radionuclides, including all positron-emitting radionuclides, to be imaged in vivo using sensitive CCD cameras. Cerenkov luminescence may also provide a means to deliver light deep inside tissue over a sustained period of time using targeted radiotracers. This light could be used for photoactivation, including photorelease of therapeutics, photodynamic therapy and photochemical internalization. Essential to assessing the feasibility of these concepts, and the design of instrumentation designed for detecting Cerenkov radiation, is an understanding of the light yield of different radionuclides in tissue. This is complicated by the dependence of the light yield on refractive index and the volume of the sample being interrogated. Using Monte Carlo simulations, in conjunction with step-wise use of the Frank-Tamm equation, we studied forty-seven different radionuclides and show that Cerenkov light yields in tissue can be as high as a few tens of photons per nuclear decay for a wavelength range of 400-800 nm. The dependency on refractive index and source volume is explored, and an expression for the scaling factor necessary to compute the Cerenkov yield in any arbitrary spectral band is given. This data will be of broad utility in guiding the application of Cerenkov radiation emitted from biomedical radionuclides.

Evaluation of Matrix9 silicon photomultiplier array for small-animal PET.

PURPOSE: The MatrixSL-9-30035-OEM (Matrix9) from SensL is a large-area silicon photomultiplier (SiPM) photodetector module consisting of a 3 × 3 array of 4 × 4 element SiPM arrays (total of 144 SiPM pixels) and incorporates SensL's front-end electronics board and coincidence board. Each SiPM pixel measures 3.16 × 3.16 mm(2) and the total size of the detector head is 47.8 × 46.3 mm(2). Using 8 × 8 polished LSO/LYSO arrays (pitch 1.5 mm) the performance of this detector system (SiPM array and readout electronics) was evaluated with a view for its eventual use in small-animal positron emission tomography (PET). METHODS: Measurements of noise, signal, signal-to-noise ratio, energy resolution, flood histogram quality, timing resolution, and array trigger error were obtained at different bias voltages (28.0-32.5 V in 0.5 V intervals) and at different temperatures (5 °C-25 °C in 5 °C degree steps) to find the optimal operating conditions. RESULTS: The best measured signal-to-noise ratio and flood histogram quality for 511 keV gamma photons were obtained at a bias voltage of 30.0 V and a temperature of 5 °C. The energy resolution and timing resolution under these conditions were 14.2% ± 0.1% and 4.2 ± 0.1 ns, respectively. The flood histograms show that all the crystals in the 1.5 mm pitch LSO array can be clearly identified and that smaller crystal pitches can also be resolved. Flood histogram quality was also calculated using different center of gravity based positioning algorithms. Improved and more robust results were achieved using the local 9 pixels for positioning along with an energy offset calibration. To evaluate the front-end detector readout, and multiplexing efficiency, an array trigger error metric is introduced and measured at different lower energy thresholds. Using a lower energy threshold greater than 150 keV effectively eliminates any mispositioning between SiPM arrays. CONCLUSIONS: In summary, the Matrix9 detector system can resolve high-resolution scintillator arrays common in small-animal PET with adequate energy resolution and timing resolution over a large detector area. The modular design of the Matrix9 detector allows it to be used as a building block for simple, low channel-count, yet high performance, small animal PET or PET/MRI systems.

Un-collimated single-photon imaging system for high-sensitivity small animal and plant imaging.

In preclinical single-photon emission computed tomography (SPECT) system development the primary objective has been to improve spatial resolution by using novel parallel-hole or multi-pinhole collimator geometries. However, such high-resolution systems have relatively poor sensitivity (typically 0.01-0.1%). In contrast, a system that does not use collimators can achieve very high-sensitivity. Here we present a high-sensitivity un-collimated detector single-photon imaging (UCD-SPI) system for the imaging of both small animals and plants. This scanner consists of two thin, closely spaced, pixelated scintillator detectors that use NaI(Tl), CsI(Na), or BGO. The performance of the system has been characterized by measuring sensitivity, spatial resolution, linearity, detection limits, and uniformity. With (99m)Tc (140 keV) at the center of the field of view (20 mm scintillator separation), the sensitivity was measured to be 31.8% using the NaI(Tl) detectors and 40.2% with CsI(Na). The best spatial resolution (FWHM when the image formed as the geometric mean of the two detector heads, 20 mm scintillator separation) was 19.0 mm for NaI(Tl) and 11.9 mm for CsI(Na) at 140 keV, and 19.5 mm for BGO at 1116 keV, which is somewhat degraded compared to the cm-scale resolution obtained with only one detector head and a close source. The quantitative accuracy of the system's linearity is better than 2% with detection down to activity levels of 100 nCi. Two in vivo animal studies (a renal scan using (99m)Tc MAG-3 and a thyroid scan with (123)I) and one plant study (a (99m)TcO4(-) xylem transport study) highlight the unique capabilities of this UCD-SPI system. From the renal scan, we observe approximately a one thousand-fold increase in sensitivity compared to the Siemens Inveon SPECT/CT scanner. UCD-SPI is useful for many imaging tasks that do not require excellent spatial resolution, such as high-throughput screening applications, simple radiotracer uptake studies in tumor xenografts, dynamic studies where very good temporal resolution is critical, or in planta imaging of radioisotopes at low concentrations.

In vivo Cerenkov luminescence imaging: a new tool for molecular imaging.

Cerenkov radiation is a phenomenon where optical photons are emitted when a charged particle moves faster than the speed of light for the medium in which it travels. Recently, we and others have discovered that measurable visible light due to the Cerenkov effect is produced in vivo following the administration of ß-emitting radionuclides to small animals. Furthermore, the amounts of injected activity required to produce a detectable signal are consistent with small-animal molecular imaging applications. This surprising observation has led to the development of a new hybrid molecular imaging modality known as Cerenkov luminescence imaging (CLI), which allows the spatial distribution of biomolecules labelled with ß-emitting radionuclides to be imaged in vivo using sensitive charge-coupled device cameras. We review the physics of Cerenkov radiation as it relates to molecular imaging, present simulation results for light intensity and spatial distribution, and show an example of CLI in a mouse cancer model. CLI allows many common radiotracers to be imaged in widely available in vivo optical imaging systems, and, more importantly, provides a pathway for directly imaging ß(-)-emitting radionuclides that are being developed for therapeutic applications in cancer and that are not readily imaged by existing methods.

Simultaneous PET and multispectral 3-dimensional fluorescence optical tomography imaging system.

UNLABELLED: Integrated PET and 3-dimensional (3D) fluorescence optical tomography (FOT) imaging has unique and attractive features for in vivo molecular imaging applications. We have designed, built, and evaluated a simultaneous PET and 3D FOT system. The design of the FOT system is compatible with many existing small-animal PET scanners. METHODS: The 3D FOT system comprises a novel conical mirror that is used to view the whole-body surface of a mouse with an electron-multiplying charge-coupled device camera when a collimated laser beam is projected on the mouse to stimulate fluorescence. The diffusion equation was used to model the propagation of optical photons inside the mouse body, and 3D fluorescence images were reconstructed iteratively from the fluorescence intensity measurements measured from the surface of the mouse. Insertion of the conical mirror into the gantry of a small-animal PET scanner allowed simultaneous PET and 3D FOT imaging. RESULTS: The mutual interactions between PET and 3D FOT were evaluated experimentally. PET has negligible effects on 3D FOT performance. The inserted conical mirror introduces a reduction in the sensitivity and noise-equivalent count rate of the PET system and increases the scatter fraction. PET-FOT phantom experiments were performed. An in vivo experiment using both PET and FOT was also performed. CONCLUSION: Phantom and in vivo experiments demonstrate the feasibility of simultaneous PET and 3D FOT imaging. The first in vivo simultaneous PET-FOT results are reported.

New covalent capture probes for imaging and therapy, based on a combination of binding affinity and disulfide bond formation.

We describe the synthesis and development of new reactive DOTA-metal complexes for covalently targeting engineered receptors in vivo, which have superior tumor uptake and clearance properties for biomedical applications. These probes are found to clear efficiently through the kidneys and minimally through other routes, but bind persistently in the tumor target. We also explore the new technique of Cerenkov luminescence imaging to optically monitor radiolabeled probe distribution and kinetics in vivo. Cerenkov luminescence imaging uniquely enables sensitive noninvasive in vivo imaging of a ß(-) emitter such as (90)Y with an optical imager.

Cerenkov luminescence tomography for small-animal imaging.

Cerenkov radiation is a well-known phenomenon in which optical photons are emitted by charged particles moving faster than the speed of light in a medium. We have observed Cerenkov photons emitted from beta-emitting radiotracers such as (18)F-fluorodeoxyglucose using a sensitive CCD camera. Phantom and in vivo mouse imaging experiments have demonstrated that surface measurements of the emitted Cerenkov optical photons could be used to reconstruct the radiotracer activity distribution inside an object by modeling the optical photon propagation with the diffusion equation and reconstructing the optical emission source distribution iteratively with a preconditioned conjugate gradient method.

A three-dimensional multispectral fluorescence optical tomography imaging system for small animals based on a conical mirror design.

We have developed a three dimensional (3D) multispectral fluorescence optical tomography small animal imaging system with an innovative geometry using a truncated conical mirror, allowing simultaneous viewing of the entire surface of the animal by an EMCCD camera. A conical mirror collects photons approximately three times more efficiently than a flat mirror. An x-y mirror scanning system makes it possible to scan a collimated excitation laser beam to any location on the mouse surface. A pattern of structured light incident on the small animal surface is used to extract the surface geometry for reconstruction. A finite element based algorithm is applied to model photon propagation in the turbid media and a preconditioned conjugate gradient (PCG) method is used to solve the large linear system matrix. The reconstruction algorithm and the system feasibility are evaluated by phantom experiments. These experiments show that multispectral measurements improve the spatial resolution of reconstructed images. Finally, an in vivo imaging study of a xenograft tumor in a mouse shows good correlation of the reconstructed image with the location of the fluorescence probe as determined by subsequent optical imaging of cryosections of the mouse.

A high-sensitivity small animal SPECT system.

Medical imaging using single gamma-ray-emitting radionuclides typically makes use of parallel hole collimators or pinholes in order to achieve good spatial resolution. However, a tradeoff in sensitivity is inherent in the use of a collimator, and modern preclinical single photon emission computed tomography (SPECT) systems detect a very small fraction of emitted gamma rays, often less than 0.1%. A system for small animal SPECT imaging which uses no collimators could potentially achieve very high sensitivity-several tens of percent-with reasonably sized detectors. This would allow two significant improvements in preclinical studies: images could be obtained more rapidly, allowing higher throughput for screening applications, or for dynamic processes to be observed with very good time resolution; and images could be obtained with less radioactive tracer, making possible the in vivo imaging of low-capacity receptor systems, aiding research into new tracer compounds, and reducing the cost and easing the regulatory burden of an experiment. Of course, a system with no collimator will not be able to approach the submillimeter spatial resolutions produced by the most advanced pinhole and collimated systems, but a high-sensitivity system with resolution of order 1 cm could nonetheless find significant and new use in the many molecular imaging applications which do not require good spatial resolution-for example, screening applications for drug development or new imaging agents. Rather than as an alternative to high-resolution SPECT systems, the high-sensitivity system is proposed as a radiotracer alternative to optical imaging for small animals. We have developed a prototype system for mouse imaging applications. The scanner consists of two large, thin, closely spaced scintillation detectors. Simulation studies indicate that a FWHM spatial resolution of 7 mm is possible. In an in vivo mouse imaging study using the (99m)Tc labeled tracer MAG-3, the sensitivity of the system is measured to be 40%. Simple projection images created by analytically combining the two detectors' data show sufficient resolution to observe the dynamic distribution of the radiotracer in the mouse.