A novel proton-integrating radiography system design using a monolithic scintillator detector: experimental studies.

Research on proton-based imaging systems aims to improve treatment planning, internal anatomy visualization, and patient alignment for proton radiotherapy. The purpose of this study was to demonstrate a new proton radiography system design consisting of a monolithic plastic scintillator volume and two optical cameras for use with scanning proton pencil beams. Unlike the thin scintillating plates currently used for proton radiography, the plastic scintillator volume (20 × 20 × 20 cm3) captures a wider distribution of proton beam energy depositions and avoids proton-beam modulation. The proton imaging system's characteristics were tested using image uniformity (2.6% over a 5 × 5 cm2 area), stability (0.37%), and linearity (R2 = 1) studies. We used the light distribution produced within the plastic scintillator to generate proton radiographs via two different approaches: (a) integrating light by using a camera placed along the beam axis, and (b) capturing changes to the proton Bragg peak positions with a camera placed perpendicularly to the beam axis. The latter method was used to plot and evaluate relative shifts in percentage depth light (PDL) profiles of proton beams with and without a phantom in the beam path. A curvelet minimization algorithm used differences in PDL profiles to reconstruct and refine the phantom water-equivalent thickness (WET) map. Gammex phantoms were used to compare the proton radiographs generated by these two methods. The relative accuracies in calculating WET of the phantoms using the calibration-based beam-integration (and the PDL) methods were -0.18 ± 0.35% (-0.29 ± 3.11%), -0.11 ± 0.51% (-0.15 ± 2.64%), -2.94 ± 1.20% (-0.75 ± 6.11%), and -1.65 ± 0.35% (0.36 ± 3.93%) for solid water, adipose, cortical bone, and PMMA, respectively. Further exploration of this unique multicamera-based imaging system is warranted and could lead to clinical applications that improve treatment planning and patient alignment for proton radiotherapy.

Image quality evaluation of projection- and depth dose-based approaches to integrating proton radiography using a monolithic scintillator detector.

The purpose of this study is to compare the image quality of an integrating proton radiography (PR) system, composed of a monolithic scintillator and two digital cameras, using integral lateral-dose and integral depth-dose image reconstruction techniques. Monte Carlo simulations were used to obtain the energy deposition in a 3D monolithic scintillator detector (30 × 30 × 30 cm3poly vinyl toluene organic scintillator) to create radiographs of various phantoms-a slanted aluminum cube for spatial resolution analysis and a Las Vegas phantom for contrast analysis. The light emission of the scintillator was corrected using Birks scintillation model. We compared two integrating PR methods and the expected results from an idealized proton tracking radiography system. Four different image reconstruction methods were utilized in this study: integral scintillation light projected from the beams-eye view, depth-dose based reconstruction methods both with and without optimization, and single particle tracking PR was used for reference data. Results showed that heterogeneity artifact due to medium-interface mismatch was identified from the Las Vegas phantom simulated in air. Spatial resolution was found to be highest for single-event reconstruction. Contrast levels, ranked from best to worst, were found to correspond to particle tracking, optimized depth-dose, depth-dose, and projection-based image reconstructions. The image quality of a monolithic scintillator integrating PR system was sufficient to warrant further exploration. These results show promise for potential clinical use as radiographic techniques for visualizing internal patient anatomy during proton radiotherapy.

Ionization quenching correction for a 3D scintillator detector exposed to scanning proton beams.

The ionization quenching phenomenon in scintillators must be corrected to obtain accurate dosimetry in particle therapy. The purpose of this study was to develop a methodology for correcting camera projection measurements of a 3D scintillator detector exposed to proton pencil beams. Birks' ionization quenching model and the energy deposition by secondary electrons (EDSE) model were used to correct the light captured by a prototype 3D scintillator detector. The detector was made of a 20 cm × 20 cm × 20 cm tank filled with liquid scintillator, and three cameras. The detector was exposed to four proton-beam energies (84.6, 100.9, 144.9, and 161.6 MeV) at The University of Texas MD Anderson Cancer Center's Proton Therapy Center. The dose and track averaged linear energy transfer (LET) were obtained using validated Monte Carlo (MC) simulations. The corrected light output was compared to the dose calculated by the MC simulation. Optical artefact corrections were used to correct for refraction at the air-scintillator interface, and image perspective. These corrections did not account for the non-orthogonal integration of data off the central axis of the image. Therefore, we compared the light output to an integrated MC dose and LET along the non-orthogonal path. After accounting for the non-orthogonal integration of the data, the corrected light output reduced the dose error at the Bragg peak region from 15% to 3% for low proton-beam energies. Overall, the doses at the Bragg peak region using the Birks' model and EDSE model were less than ±3% and ±7% of the MC dose, respectively. We have improved the application of Birks' model quenching corrections in 3D scintillators by numerically projecting the dose and LET 3D grid to camera projections. This study shows that scintillator projections can be corrected using average LET values at the central axes.

A proton imaging system using a volumetric liquid scintillator: a preliminary study.

With the expansion of proton radiotherapy for cancer treatments, it has become important to explore proton-based imaging technologies to increase the accuracy of proton treatment planning, alignment, and verification. The purpose of this study is to demonstrate the feasibility of using a volumetric liquid scintillator to generate proton radiographs at a clinically relevant energy (180 MeV) using an integrating detector approach. The volumetric scintillator detector is capable of capturing a wide distribution of residual proton beam energies from a single beam irradiation. It has the potential to reduce acquisition time and imaging dose compared to other proton radiography methods. The imaging system design is comprised of a volumetric (20 × 20 × 20 cm3) organic liquid scintillator working as a residual-range detector and a charge-coupled device (CCD) placed along the beams'-eye-view for capturing radiographic projections. The scintillation light produced within the scintillator volume in response to a 3-dimensional distribution of residual proton beam energies is captured by the CCD as a 2-dimensional grayscale image. A light intensity-to-water equivalent thickness (WET) curve provided WET values based on measured light intensities. The imaging properties of the system, including its contrast, signal-to-noise ratio, and spatial resolution (0.19 line-pairs/mm) were determined. WET values for selected Gammex phantom inserts including solid water, acrylic, and cortical bone were calculated from the radiographs with a relative accuracy of -0.82%, 0.91%, and -2.43%, respectively. Image blurring introduced by system optics was accounted for, resulting in sharper image features. Finally, the system's ability to reconstruct proton CT images from radiographic projections was demonstrated using a filtered back-projection algorithm. The WET retrieved from the reconstructed CT slice was within 0.3% of the WET obtained from MC. In this work, the viability of a cumulative approach to proton imaging using a volumetric liquid scintillator detector and at a clinically-relevant energy was demonstrated.

Performance characterization of a 3D liquid scintillation detector for discrete spot scanning proton beam systems.

Existing systems for proton beam dosimetry are limited in their ability to provide a complete, accurate, and detailed account of volumetric dose distribution. In this work, we describe the design and development of a portable, fast, and reusable liquid scintillator-based three-dimensional (3D) optical detection system for use in proton therapy. Our long-term goal is to use this system clinically for beam characterization, dosimetry, and quality assurance studies of discrete spot scanning proton beam systems. The system used a 20 × 20 × 20 cm3 liquid scintillator volume. Three mutually orthogonal cameras surrounding this volume captured scintillation photons emitted in response to the proton beams. The cameras exhibited a mean spatial resolution of 0.21 mm over the complete detection volume and a temporal resolution of 11 ms. The system is shown to be capable of capturing all 94 beam energies delivered by a synchrotron and performing rapid beam range measurements with a mean accuracy of 0.073 ± 0.030 mm over all energies. The range measurement uncertainty for doses less than 1 cGy was found to be ±0.355 mm, indicating high precision for low dose detection. Finally, we demonstrated that using multiple cameras allowed for the precise locations of the delivered beams to be tracked in 3D. We conclude that this detector is capable of real-time and accurate tracking of dynamic spot beam deliveries in 3D. The high-resolution light profiles it generates will be useful for future 3D construction of dose maps.

Lymphatic transport in patients with chronic venous insufficiency and venous leg ulcers following sequential pneumatic compression.

BACKGROUND: Recent advancements in near-infrared fluorescence lymphatic imaging (NIRFLI) technology provide opportunities for non-invasive, real-time assessment of lymphatic contribution in the etiology and treatment of ulcers. The objective of this study was to assess lymphatics in subjects with venous leg ulcers using NIRFLI and to assess lymphatic impact of a single session of sequential pneumatic compression (SPC). METHODS: Following intradermal microdoses of indocyanine green (ICG) as a lymphatic contrast agent, NIRFLI was used in a pilot study to image the lymphatics of 12 subjects with active venous leg ulcers (Clinical, Etiologic, Anatomic, and Pathophysiologic [CEAP] C6). The lymphatics were imaged before and after a single session of SPC to assess impact on lymphatic function. RESULTS: Baseline imaging showed impaired lymphatic function and bilateral dermal backflow in all subjects with chronic venous insufficiency, even those without ulcer formation in the contralateral limb (C0 and C4 disease). SPC therapy caused proximal movement of ICG away from the active wound in 9 of 12 subjects, as indicated by newly recruited functional lymphatic vessels, emptying of distal lymphatic vessels, or proximal movement of extravascular fluid. Subjects with the longest duration of active ulcers had few visible lymphatic vessels, and proximal movement of ICG was not detected after SPC therapy. CONCLUSIONS: This study provides visible confirmation of lymphatic dysfunction at an early stage in the etiology of venous ulcer formation and demonstrates the potential therapeutic mechanism of SPC therapy in removing excess fluid. The ability of SPC therapy to restore fluid balance through proximal movement of lymph and interstitial fluid may explain its value in hastening venous ulcer healing. Anatomical differences between the lymphatics of longstanding and more recent venous ulcers may have important therapeutic implications.

Experimental Comparison of Continuous-Wave and Frequency-Domain Fluorescence Tomography in a Commercial Multi-Modal Scanner.

The performance evaluation of a variety of small animal tomography measurement approaches and algorithms for recovery of fluorescent absorption cross section has not been conducted. Herein, we employed an intensified CCD system installed in a commercial small animal CT (Computed Tomography) scanner to compare image reconstructions from time-independent, continuous wave (CW) measurements and from time-dependent, frequency domain (FD) measurements in a series of physical phantoms specifically designed for evaluation. Comparisons were performed as a function of (1) number of projections, (2) the level of preprocessing filters used to improve the signal-to-noise ratio (SNR), (3) endogenous heterogeneity of optical properties, as well as in the cases of (4) two fluorescent targets and (5) a mouse-shaped phantom. Assessment of quantitative recovery of fluorescence absorption cross section was performed using a fully parallel, regularization-free, linear reconstruction algorithm with diffusion approximation (DA) and high order simplified spherical harmonics ( SPN) approximation to the radiative transport equation (RTE). The results show that while FD measurements may result in superior image reconstructions over CW measurements, data acquisition times are significantly longer, necessitating further development of multiple detector/source configurations, improved data read-out rates, and detector technology. FD measurements with SP3 reconstructions enabled better quantitative recovery of fluorescent target strength, but required increased computational expense. Despite the developed parallel reconstruction framework being able to achieve more than 60 times speed increase over sequential implementation, further development in faster parallel acceleration strategies for near-real time and real-time image recovery and more precise forward solution is necessary.

Evidence for SH2 domain-containing 5'-inositol phosphatase-2 (SHIP2) contributing to a lymphatic dysfunction.

The lymphatic vasculature plays a critical role in a number of disease conditions of increasing prevalence, such as autoimmune disorders, obesity, blood vascular diseases, and cancer metastases. Yet, unlike the blood vasculature, the tools available to interrogate the molecular basis of lymphatic dysfunction/disease have been lacking. More recently, investigators have reported that dysregulation of the PI3K pathway is involved in syndromic human diseases that involve abnormal lymphatic vasculatures, but there have been few compelling results that show the direct association of this molecular pathway with lymphatic dysfunction in humans. Using near-infrared fluorescence lymphatic imaging (NIRFLI) to phenotype and next generation sequencing (NGS) for unbiased genetic discovery in a family with non-syndromic lymphatic disease, we discovered a rare, novel mutation in INPPL1 that encodes the protein SHIP2, which is a negative regulator of the PI3K pathway, to be associated with lymphatic dysfunction in the family. In vitro interrogation shows that SHIP2 is directly associated with impairment of normal lymphatic endothelial cell (LEC) behavior and that SHIP2 associates with receptors that are associated in lymphedema, implicating its direct involvement in the lymphatic vasculature.

An abnormal lymphatic phenotype is associated with subcutaneous adipose tissue deposits in Dercum's disease.

OBJECTIVE: Investigational, near-infrared fluorescence (NIRF) lymphatic imaging was used to assess lymphatic architecture and contractile function in participants diagnosed with Dercum's disease, a rare, poorly understood disorder characterized by painful lipomas in subcutaneous adipose tissues. METHODS: After informed consent and as part of an FDA-approved feasibility study to evaluate lymphatics in diseases in which their contribution has been implicated, three women diagnosed with Dercum's disease and four control subjects were imaged. Each participant received multiple intradermal and subcutaneous injections of indocyanine green (ICG, total dose ≤400 µg) in arms, legs, and/or trunk. Immediately after injection, ICG was taken up by the lymphatics and NIRF imaging was conducted. RESULTS: The lymphatics in the participants with Dercum's disease were intact and dilated, yet sluggishly propelled lymph when compared to control lymphatics. Palpation of regions containing fluorescent lymphatic pathways revealed tender, fibrotic, tubular structures within the subcutaneous adipose tissue that were associated with painful nodules, and, in some cases, masses of fluorescent tissue indicating that some lipomas may represent tertiary lymphoid tissues. CONCLUSIONS: These data support the hypothesis that Dercum's disease may be a lymphovascular disorder and suggest a possible association between abnormal adipose tissue deposition and abnormal lymphatic structure and function.

Small animal fluorescence and bioluminescence tomography: a review of approaches, algorithms and technology update.

Emerging fluorescence and bioluminescence tomography approaches have several common, yet several distinct features from established emission tomographies of PET and SPECT. Although both nuclear and optical imaging modalities involve counting of photons, nuclear imaging techniques collect the emitted high energy (100-511 keV) photons after radioactive decay of radionuclides while optical techniques count low-energy (1.5-4.1 eV) photons that are scattered and absorbed by tissues requiring models of light transport for quantitative image reconstruction. Fluorescence imaging has been recently translated into clinic demonstrating high sensitivity, modest tissue penetration depth, and fast, millisecond image acquisition times. As a consequence, the promise of quantitative optical tomography as a complement of small animal PET and SPECT remains high. In this review, we summarize the different instrumentation, methodological approaches and schema for inverse image reconstructions for optical tomography, including luminescence and fluorescence modalities, and comment on limitations and key technological advances needed for further discovery research and translation.

Far-red fluorescence gene reporter tomography for determination of placement and viability of cell-based gene therapies.

Non-invasive injectable cellular therapeutic strategies based on sustained delivery of physiological levels of BMP-2 for spinal fusion are emerging as promising alternatives, which could provide sufficient fusion without the associated surgical risks. However, these injectable therapies are dependent on bone formation occurring only at the specific target region. In this study, we developed and deployed fluorescence gene reporter tomography (FGRT) to provide information on in vivo cell localization and viability. This information is sought to confirm the ideal placement of the materials with respect to the area where early bone reaction is required, ultimately providing three dimensional data about the future fusion. However, because almost all conventional fluorescence gene reporters require visible excitation wavelengths, current in vivo imaging of fluorescent proteins is limited by high tissue absorption and confounding autofluorescence. We previously administered fibroblasts engineered to produce BMP-2, but is difficult to determine 3-D information of placement prior to bone formation. Herein we used the far-red fluorescence gene reporter, IFP1.4 to report the position and viability of fibroblasts and developed 3-D tomography to provide placement information. A custom small animal, far-red fluorescence tomography system integrated into a commercial CT scanner was used to assess IFP1.4 fluorescence and to demark 3-D placement of encapsulated fibroblasts with respect to the vertebrae and early bone formation as assessed from CT. The results from three experiments showed that the placement of the materials within the spine could be detected. This work shows that in vivo fluorescence gene reporter tomography of cell-based gene therapy is feasible and could help guide cell-based therapies in preclinical models.

In vivo imaging of orthotopic prostate cancer with far-red gene reporter fluorescence tomography and in vivo and ex vivo validation.

Fluorescence gene reporters have recently become available for excitation at far-red wavelengths, enabling opportunities for small animal in vivo gene reporter fluorescence tomography (GRFT). We employed multiple projections of the far-red fluorescence gene reporters IFP1.4 and iRFP, excited by a point source in transillumination geometry in order to reconstruct the location of orthotopically implanted human prostate cancer (PC3), which stably expresses the reporter. Reconstruction was performed using a linear radiative-transfer-based regularization-free tomographic method. Positron emission tomography (PET) imaging of a radiolabeled antibody-based agent that targeted epithelial cell adhesion molecule overexpressed on PC3 cells was used to confirm in vivo GRFT results. Validation of GRFT results was also conducted from ex vivo fluorescence imaging of resected prostate tumor. In addition, in mice with large primary prostate tumors, a combination of GRFT and PET showed that the radiolabeled antibody did not penetrate the tumor, consistent with known tumor transport limitations of large (∼150 kDa) molecules. These results represent the first tomography of a living animal using far-red gene reporters.

A compact frequency-domain photon migration system for integration into commercial hybrid small animal imaging scanners for fluorescence tomography.

The work presented herein describes the system design and performance evaluation of a miniaturized near-infrared fluorescence (NIRF) frequency-domain photon migration (FDPM) system with non-contact excitation and homodyne detection capability for small animal fluorescence tomography. The FDPM system was developed specifically for incorporation into a Siemens micro positron emission tomography/computed tomography (microPET/CT) commercial scanner for hybrid small animal imaging, but could be adapted to other systems. Operating at 100 MHz, the system noise was minimized and the associated amplitude and phase errors were characterized to be ±0.7% and ±0.3°, respectively. To demonstrate the tomographic ability, a commercial mouse-shaped phantom with 50 µM IRDye800CW and 68Ga containing inclusion was used to associate PET and NIRF tomography. Three-dimensional mesh generation and anatomical referencing was accomplished through CT. A third-order simplified spherical harmonics approximation (SP3) algorithm, for efficient prediction of light propagation in small animals, was tailored to incorporate the FDPM approach. Finally, the PET-NIRF target co-localization accuracy was analyzed in vivo with a dual-labeled imaging agent targeting orthotopic growth of human prostate cancer. The obtained results validate the integration of time-dependent fluorescence tomography system within a commercial microPET/CT scanner for multimodality small animal imaging.

Improvement of fluorescence-enhanced optical tomography with improved optical filtering and accurate model-based reconstruction algorithms.

The goal of preclinical fluorescence-enhanced optical tomography (FEOT) is to provide three-dimensional fluorophore distribution for a myriad of drug and disease discovery studies in small animals. Effective measurements, as well as fast and robust image reconstruction, are necessary for extensive applications. Compared to bioluminescence tomography (BLT), FEOT may result in improved image quality through higher detected photon count rates. However, background signals that arise from excitation illumination affect the reconstruction quality, especially when tissue fluorophore concentration is low and/or fluorescent target is located deeply in tissues. We show that near-infrared fluorescence (NIRF) imaging with an optimized filter configuration significantly reduces the background noise. Model-based reconstruction with a high-order approximation to the radiative transfer equation further improves the reconstruction quality compared to the diffusion approximation. Improvements in FEOT are demonstrated experimentally using a mouse-shaped phantom with targets of pico- and subpico-mole NIR fluorescent dye.

Direct evidence of lymphatic function improvement after advanced pneumatic compression device treatment of lymphedema.

Lymphedema affects up to 50% of all breast cancer survivors. Management with pneumatic compression devices (PCDs) is controversial, owing to the lack of methods to directly assess benefit. This pilot study employed an investigational, near-infrared (NIR) fluorescence imaging technique to evaluate lymphatic response to PCD therapy in normal control and breast cancer-related lymphedema (BCRL) subjects. Lymphatic propulsion rate, apparent lymph velocity, and lymphatic vessel recruitment were measured before, during, and after advanced PCD therapy. Lymphatic function improved in all control subjects and all asymptomatic arms of BCRL subjects. Lymphatic function improved in 4 of 6 BCRL affected arms, improvement defined as proximal movement of dye after therapy. NIR fluorescence lymphatic imaging may be useful to directly evaluate lymphatic response to therapy. These results suggest that PCDs can stimulate lymphatic function and may be an effective method to manage BCRL, warranting future clinical trials.