Gregory S. Mitchell

Optical imaging of Cerenkov light generation from positron-emitting radiotracers

Radiotracers labeled with high-energy positron emitters, such as those commonly used for positron emission tomography studies, emit visible light immediately following decay in a medium. This phenomenon, not previously described for these imaging tracers, is consistent with Cerenkov radiation and has several potential applications, especially for in vivo molecular imaging studies. Herein we detail a new molecular imaging tool, Cerenkov Luminescence Imaging, the experiments conducted that support our interpretation of the source of the signal, and proof-of-concept in vivo studies that set the foundation for future application of this new method.

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.

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 β-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 β-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 β−-emitting radionuclides that are being developed for therapeutic applications in cancer and that are not readily imaged by existing methods.

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 hyperspectral fluorescence system for 3D in vivo optical imaging

In vivo optical instruments designed for small animal imaging generally measure the integrated light intensity across a broad band of wavelengths, or make measurements at a small number of selected wavelengths, and primarily use any spectral information to characterize and remove autofluorescence. We have developed a flexible hyperspectral imaging instrument to explore the use of spectral information to determine the 3D source location for in vivo fluorescence imaging applications. We hypothesize that the spectral distribution of the emitted fluorescence signal can be used to provide additional information to 3D reconstruction algorithms being developed for optical tomography. To test this hypothesis, we have designed and built an in vivo hyperspectral imaging system, which can acquire data from 400 to 1000 nm with 3 nm spectral resolution and which is flexible enough to allow the testing of a wide range of illumination and detection geometries. It also has the capability to generate a surface contour map of the animal for input into the reconstruction process. In this paper, we present the design of the system, demonstrate the depth dependence of the spectral signal in phantoms and show the ability to reconstruct 3D source locations using the spectral data in a simple phantom. We also characterize the basic performance of the imaging system.

CdTe Strip Detector Characterization for High Resolution Small Animal PET

Excellent spatial resolution is a requirement for preclinical PET imaging. In order to achieve spatial resolution of significantly better than one millimeter, an appealing possibility is to employ direct detector materials, such as cadmium telluride (CdTe). Prototype thin orthogonal strip detectors have been developed for testing. They have dimensions of 20 mm by 20 mm and are 0.5 mm thick, and have strips of 0.5 mm pitch on one side and 2.5 mm pitch on the other. Results are presented for the energy resolution (3% at 511 keV), intrinsic position resolution (equal to the 0.5 mm strip pitch), and timing resolution (3 ns FWHM in coincidence with an LSO detector, 8 ns FWHM for coincidence of two CdTe detectors) of the detectors. A PET scanner design is proposed using blocks made of the CdTe strip detectors, oriented in the blocks with their thin edges toward the center of the scanner. Simulation results suggest that this scanner, using a threshold of 250 keV, would have a sensitivity of 3.4% for a point source at its center.

Simultaneous PET and Multispectral 3-Dimensional Fluorescence Optical Tomography Imaging System

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.

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.

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 β(-) emitter such as (90)Y with an optical imager.

CdTe Orthogonal Strip Detector for Small Animal PET

In this paper we present experimental results obtained with an orthogonal strip CdTe detector designed for application to small animal PET. The detector was designed to provide timing resolution acceptable for small animal PET, depth of interaction information and spatial resolution superior to scintillator based detectors. A CdTe sample of 20times20 mm2 area and 0.5 mm thickness was patterned with strips of 0.5-mm pitch on one 20times20 mm2 face. In a small animal PET detector ring the 511 keV gamma-rays would be incident on the edge of the device (i.e., 0.5times20 mm2 face) and encounter 20-mm depth, providing reasonably high stopping efficiency. The strips with 0.5-mm pitch on the 20times20 mm2 face (in combination with the device thickness of 0.5-mm) would provide identification of the event in an effective pixel size of 0.5times0.5 mm2. The other 20times20 mm2 face of the device has coarser strips with 2.0-mm width (2.5-mm pitch) in the direction orthogonal to the finer 0.5-mm strips. These 2-mm strips provide the depth of interaction (DOI) for the 511 keV gamma-rays. Since the overall thickness between the electrodes is only 0.5-mm, fast and efficient collection of electrons and holes is possible, resulting in high energy and timing resolution. Pulse height spectra under 22Na irradiation, coincidence timing resolution and detector response to a collimated source are presented.