Mouse Phantom Research Articles

ElectronCT - an imaging technique using very-high energy electrons

The electronCT technique is an imaging method based on the multiple Coulomb scattering of relativistic electrons and has potential applications in medical and industrial imaging. It utilizes a pencil beam of electrons in the very high energy electron (VHEE, 50–250 MeV) range and a single detection layer for the determination of the beam profile. The technique constitutes a projectional, two-dimensional imaging method and thus also qualifies for the tomographic reconstruction of samples. Given the simplicity of the technical setup and its location behind the sample, the electronCT technique has potential synergies with VHEE radiotherapy, making use of the same electron source for both treatment and diagnostics and thus being a candidate for in situ imaging and patient localization. At the same time, several technical challenges arise from the measurement technique when applied for the imaging of living beings. Measurements performed at the ARES linear particle accelerator at an electron energy of 155 MeV using a mouse phantom and a Timepix3 silicon pixel detector assembly demonstrate the feasibility of this technique. Both projectional and tomographic reconstructions are presented and the potential and limits of the technology are discussed.

Exploring a tristhione scorpionate ligand as a suitable chelator for the theranostic pair antimony-119 and antimony-117

BackgroundThe highly potent Auger electron emitter antimony-119 (119Sb) and the SPECT-isotope antimony-117 (117Sb) comprise a true theranostic pair particularly suitable for cancer theranostics. Harnessing this potential requires development of a chelator that can rapidly form a stable complex with radioactive antimony ions at the low concentrations typical of radiopharmaceutical preparations. Stable Sb(III) complexes of hydrotris(methimazolyl)borate (TMe) are known, prompting our investigation of this chelator. Additionally, the production of radioantimony was optimized and the SPECT imaging properties of 117Sb was investigated, in an attempt to move towards biomedical implementation of the theranostic isotope pair of antimony.ResultsA method for rapid and effective labelling of TMe using 117Sb was developed, yielding high radiochemical purities of 98.5 ± 2.7% and high radionuclidic purities exceeding 99%. Radiolabelling yielded an Sb(III) complex directly from the acidic Sb(V) solution. [1XXSb]Sb-TMe in aqueous acidic solution showed high stability in the presence of cysteine, however, the stability of the radiocomplex at increased pH was significantly decreased. The production method of 117Sb was optimized, enabling a production yield of up to 19.6 MBq/µAh and the production of up to 564 MBq at end of bombardment, following irradiation of a thin 117Sn-enriched solid target. Preclinical SPECT/CT scanning of a mouse phantom containing purified 117Sb demonstrated excellent SPECT imaging properties of 117Sb with high spatial resolution comparable to that of technetium-99m.ConclusionWe have explored the TMe chelator for complexation of radioantimony and devised a rapid chelation protocol suitable for the short half-life of 117Sb (T1/2 = 2.8 h). [1XXSb]Sb-TMe (1XXSb = 117Sb, 118mSb, 120mSb and 122Sb) demonstrated a high stability in presence of cysteine, although low stability was observed at pH > 4. We have achieved a production yield of 117Sb high enough for clinical applications and demonstrated the excellent SPECT-imaging properties of 117Sb. The results contribute valuable information for the development of suitable chelators for radioantimony and is a step further towards implementation of the antimony theranostic pair in biomedical applications.

A simulation framework for preclinical proton irradiation workflow

Objective.The integration of proton beamlines with x-ray imaging/irradiation platforms has opened up possibilities for image-guided Bragg peak irradiations in small animals. Such irradiations allow selective targeting of normal tissue substructures and tumours. However, their small size and location pose challenges in designing experiments. This work presents a simulation framework useful for optimizing beamlines, imaging protocols, and design of animal experiments. The usage of the framework is demonstrated, mainly focusing on the imaging part.Approach.The fastCAT toolkit was modified with Monte Carlo (MC)-calculated primary and scatter data of a small animal imager for the simulation of micro-CT scans. The simulated CT of a mini-calibration phantom from fastCAT was validated against a full MC TOPAS CT simulation. A realistic beam model of a preclinical proton facility was obtained from beam transport simulations to create irradiation plans in matRad. Simulated CT images of a digital mouse phantom were generated using single-energy CT (SECT) and dual-energy CT (DECT) protocols and their accuracy in proton stopping power ratio (SPR) estimation and their impact on calculated proton dose distributions in a mouse were evaluated.Main results.The CT numbers from fastCAT agree within 11 HU with TOPAS except for materials at the centre of the phantom. Discrepancies for central inserts are caused by beam hardening issues. The root mean square deviation in the SPR for the best SECT (90 kV/Cu) and DECT (50 kV/Al-90 kV/Al) protocols are 3.7% and 1.0%, respectively. Dose distributions calculated for SECT and DECT datasets revealed range shifts <0.1 mm, gamma pass rates (3%/0.1 mm) greater than 99%, and no substantial dosimetric differences for all structures. The outcomes suggest that SECT is sufficient for proton treatment planning in animals.Significance.The framework is a useful tool for the development of an optimized experimental configuration without using animals and beam time.

Contrast-to-noise ratio comparison between X-ray fluorescence emission tomography and computed tomography.

We provide a comparison of X-ray fluorescence emission tomography (XFET) and computed tomography (CT) for detecting low concentrations of gold nanoparticles (GNPs) in soft tissue and characterize the conditions under which XFET outperforms energy-integrating CT (EICT) and photon-counting CT (PCCT). We compared dose-matched Monte Carlo XFET simulations and analytical fan-beam EICT and PCCT simulations. Each modality was used to image a numerical mouse phantom and contrast-depth phantom containing GNPs ranging from 0.05% to 4% by weight in soft tissue. Contrast-to-noise ratios (CNRs) of gold regions were compared among the three modalities, and XFET's detection limit was quantified based on the Rose criterion. A partial field-of-view (FOV) image was acquired for the phantom region containing 0.05% GNPs. For the mouse phantom, XFET produced superior CNR values ( , 21.6, and 3.4) compared with CT images obtained with both energy-integrating ( , 4.6, and 1.5) and photon-counting ( , 7.7, and 2.0) detection systems. More generally, XFET outperformed CT for superficial imaging depths ( ) for gold concentrations at and above 0.5%. XFET's surface detection limit was quantified as 0.44% for an average phantom dose of 16mGy compatible with in vivo imaging. XFET's ability to image partial FOVs was demonstrated, and 0.05% gold was easily detected with an estimated dose of to a localized region of interest. We demonstrate a proof of XFET's benefit for imaging low concentrations of gold at superficial depths and the feasibility of XFET for in vivo metal mapping in preclinical imaging tasks.

Three-Dimensional-Bioprinted Non-Small Cell Lung Cancer Models in a Mouse Phantom for Radiotherapy Research.

Lung cancer continues to have one of the highest morbidity and mortality rates of any cancer. Although radiochemotherapy, in combination with immunotherapy, has significantly improved overall survival, new treatment options are urgently needed. However, preclinical radiotherapy testing is often performed in animal models, which has several drawbacks, including species-specific differences and ethical concerns. To replace animal models, this study used a micro-extrusion bioprinting approach to generate a three-dimensional (3D) human lung cancer model consisting of lung tumor cells embedded in human primary lung fibroblasts for radiotherapy research. The models were placed in a mouse phantom, i.e., a 3D-printed mouse model made of materials that mimic the X-ray radiation attenuation rates found in mice. In radiotherapy experiments, the model demonstrated a selective cytotoxic effect of X-rays on tumor cells, consistent with findings in 2D cells. Furthermore, the analysis of metabolic activity, cell death, apoptosis, and DNA damage-induced γH2AX foci formation revealed different results in the 3D model inside the phantom compared to those observed in irradiated models without phantom and 2D cells. The proposed setup of the bioprinted 3D lung model inside the mouse phantom provides a physiologically relevant model system to study radiation effects.

Development and validation of a comprehensive S-value database for small animal internal dosimetry in nuclear medicine using the DM_Bra mouse phantom

Animal models are essential in the development of new radiopharmaceuticals in nuclear medicine, particularly for accurate dose calculation in small animal internal dosimetry. This study presents a comprehensive dataset of S-values for eleven commonly used radionuclides, calculated using the DM_Bra mouse phantom with the GATE Monte Carlo simulation code. To validate our approach, we first compared S-values obtained from the DM_Bra phantom with published values derived from the Digimouse phantom using a Tc-99 m source. The differences between the two phantoms range from 0.68% to 12.45% for self-irradiation and from 0.15% to 4.19% for cross-irradiation when the source is the stomach. These results demonstrate good agreement with reference data, supporting the reliability of our dataset. We then expanded our analysis by generating S-values for additional radionuclides, reflecting their usage in both diagnostic and therapeutic applications. Furthermore, to assess the impact of varying mouse geometries on S-values, the DM_Bra phantom (26.9 g) was rescaled to represent two other mouse sizes (19.6 g and 35.9 g). The statistical uncertainty associated with all these S-values remains below 2%. This study offers a valuable resource for internal dosimetry in mice, providing detailed S-values for a wide range of radionuclides and organ geometries, which can be used in small animal PET and SPECT studies.

Commissioning an ultra-high-dose-rate electron linac with end-to-end tests

Objective. The FLASH effect can potentially be used to improve the therapeutic ratio of radiotherapy (RT) through delivery of Ultra-high-dose-rate (UHDR) irradiation. Research is actively being conducted to translate UHDR-RT and for this purpose the Mobetron is capable of producing electron beams at both UHDR and conventional dose rates for FLASH research and translation. This work presents commissioning of an UHDR Mobetron with end-to-end tests developed for preclinical research. Approach. UHDR electron beams were commissioned with an efficient approach utilizing a 3D-printed water tank and film to fully characterize beam characteristics and dependences on field size, pulse width (PW) and pulse repetition frequency (PRF). This commissioning data was used to implement a beam model using the GAMOS Monte Carlo toolkit for the preclinical research. Then, the workflow for preclinical FLASH irradiation was validated with end-to-end tests delivered to a 3D-printed mouse phantom with internal inhomogeneities. Main results. PDDs, profiles and output factors acquired with radiochromic films were precisely measured, with a PRF that showed little effect on the UHDR beam energy and spatial characteristics. Increasing PW reduced the D max and R50 by 2.08 mm µs−1 and 1.28 mm µs−1 respectively. An end-to-end test of the preclinical research workflow showed that both profiles in head-foot and lateral directions were in good agreement with the MC calculations for the heterogeneous 3D printed mouse phantom with Gamma index above 93% for 2 mm/2% criteria, and 99% for 3 mm/3%. Significance. The UHDR Mobetron is a versatile tool for FLASH preclinical research and this comprehensive beam model and workflow was validated to meet the requirements for conducting translational FLASH research.

The feasibility of K XRF bone lead measurements in mice assessed using 3D-printed phantoms

This article describes the development of a system for in vivo measurements of lead body burden in mice using 109Cd K x-ray fluorescence (XRF). This K XRF system could facilitate early-stage studies on interventions that ameliorate or reverse organ tissue damage from lead poisoning by reducing animal numbers through a cross-sectional study approach. A novel mouse phantom was developed based on a mouse atlas and 3D-printed using PLA plastic with plaster of Paris ’bone’ inserts. PLA plastic was found to be a good surrogate for soft tissue in XRF measurements and the phantoms were found to be good models of mice. As expected, lead detection limits varied with mouse size, mouse orientation, and mouse position with respect to the source and detector. The work suggests that detection limits of 10 to 20 μg Pb per g bone mineral may be possible for a 2 to 3 hour XRF measurement in a single animal, an adequate limit for some pre-clinical studies. The 109Cd K XRF mouse measurement system was also modeled using the Monte Carlo code MCNP. The combination of experiment and modeling found that contrary to expectation, accurate measurements of lead levels in mice required calibration using mouse-specific calibration standards due to the coherent scatter peak normalization failing when small animals are measured. MCNP modeling determined that this was because the coherent scatter signal from soft tissue, which until now has been assumed negligible, becomes significant when compared to the coherent scatter signal in bone in small animals. This may have implications for some human measurements. This work suggests that 109Cd K x-ray fluorescence measurements of lead body burden are precise enough to make the system feasible for small animals if appropriately calibrated. Further work to validate the technology’s measurement accuracy and performance in vivo will be required.

Detection of gastrointestinal foreign bodies in pets using single-grid-based dark-field X-ray imaging

Gastrointestinal foreign bodies (GI-FBs) occur when pets consume non-digestible items that do not readily pass through the stomach or intestines. While some GI-FBs pass through, many become lodged along the gastrointestinal tract and cause discomfort, often leading to sickness. Conventional absorption-based radiography is widely used to detect GI-FBs in pets. However, detecting low-density FBs, such as wood, plastic, clothing, and sticks, is typically difficult using conventional radiography. This study presents a novel approach for detecting low-density GI-FBs in pets by using single-grid-based dark-field X-ray imaging (SG-DFXI). It obtains microstructural information from small-angle X-ray scattering (SAXS) of the sample using a conventional X-ray source and grid. SG-DFXI requires minimal exposure and system setup and is specifically utilized to detect low-density materials that are invisible when using conventional radiography. To validate the efficacy of the proposed method, an experiment was conducted on a mouse phantom attached to a wooden chopstick as a low-density GI-FB. Quantitative evaluation was performed using image quality metrics of the contrast-to-noise ratio (CNR) and relative contrast gain (RCG). The CNR value measured in the dark-field image obtained with an autocorrelation length of ξG = 294 nm was 4.59, approximately 5.4 times larger than that of the absorption image obtained in the same imaging setup. In addition, the RCG characteristics improved as the autocorrelation length of the system increased; the RCG value for ξG = 294 nm was 5.4, approximately 3.2 times larger than that for ξG = 194 nm. Thus, increasing the autocorrelation length of the system is critical to improve its ability to detect low-density FBs. Consequently, SG-DFXI appears to be a promising method that can be used effectively and easily to detect GI-FBs in pets, which are barely visible in absorption images.

Multi-Institutional Audit of FLASH and Conventional Dosimetry With a 3D Printed Anatomically Realistic Mouse Phantom

PurposeWe conducted a multi-institutional dosimetric audit between FLASH and conventional dose rate (CONV) electron irradiations by using an anatomically realistic 3D-printed mouse phantom. MethodsA CT scan of a live mouse was used to create a 3D model of bony anatomy, lungs, and soft tissue. A dual-nozzle 3D printer was used to print the mouse phantom using acrylonitrile butadiene styrene (∼1.02 g/cm3) and polylactic acid (∼1.24 g/cm3) simultaneously to simulate soft tissue and bone densities, respectively. The lungs were printed separately using lightweight polylactic acid (∼0.64 g/cm3). Hounsfield units (HU), densities and print-to-print stability of the phantoms were assessed. Three institutions were each provided a phantom, and each institution performed two replicates of irradiations at selected anatomic regions. The average dose difference between FLASH and CONV dose distributions and deviation from the prescribed dose were measured with radiochromic film. ResultsCompared to the reference CT scan, CT scans of the phantom demonstrated mass density differences of 0.10 g/cm3 for bone, 0.12 g/cm3 for lung, and 0.03 g/cm3 for soft tissue regions. Differences in HU between phantoms were <10 HU for soft tissue and bone, with lung showing the most variation (54 HU), but with minimal impact on dose distribution (<0.5%). Mean differences between FLASH and CONV decreased from the first to the second replicate (4.3% to 1.2%), while differences from the prescribed dose decreased for both CONV (3.6% to 2.5%) and FLASH (6.4% to 2.7%). Total dose accuracy suggests consistent pulse dose and pulse number, though these were not specifically assessed. Positioning variability was observed, likely due to the absence of robust positioning aids or image guidance. ConclusionsThis study marks the first dosimetric audit for FLASH using a non-homogeneous phantom, challenging conventional calibration practices reliant on homogeneous phantoms. The comparison protocol offers a framework for credentialing multi-institutional studies in FLASH preclinical research to enhance reproducibility of biological findings.

Commissioning and Assessment of Radiation Field and Dose Inhomogeneity for a Dual X-ray Tube Cabinet Irradiator: To Ensure Accurate Dosimetry in Radiation Biology Experiments

PurposeStandardization of X-ray cabinet irradiators dose, geometry, and calibration reporting is an ongoing process. Multi-tube designs have been introduced onto the pre-clinical market and give a theoretical benefit but have not been widely assessed for use in pre-clinical irradiation conditions. The aim of this study was to report on our experience commissioning a dual x-ray source cabinet irradiator (CIXD, Xstrahl Limited, United Kingdom) and assess the dose distribution for various experimental conditions. Materials and MethodsHalf-value layer (HVL) measurement, profile measurements, and output calibration were performed with a calibrated ion chamber. Constancy measurements were performed twice daily over two weeks to assess output fluctuations. Film measurements were completed using solid water to assess percent depth dose and homogeneity within the field and within variable thickness of solid water and phosphate-buffered saline (PBS) solution. Film measurements were repeated for various arrangements of petri dishes filled with PBS or water and in a 3D printed mouse phantom. ResultsThe X-ray tubes measured in-air output was 1.27 Gy/min. The HVL was 1.7 mm Cu. The upper and lower tubes both exhibited the heel effect, but when operated simultaneously the effect was reduced. Ion chamber measurements revealed 15% dose inhomogeneity within the tray area (18 × 18 cm2). Film measurements in the petri dishes indicated minor non-uniformities with the arrangements of experimental apparatus. The mouse phantom with film agreed with ion chamber measurements for various phantom placement and orientations. ConclusionX-ray cell culture irradiation with the dual tube cabinet irradiation is efficient and robust when utilizing established dosimetric tools to confirm output and homogeneity. The conditions assumed for calibrations are often not maintained during experiments. We have confirmed that inhomogeneities are present for single tube use, however they are reduced with simultaneous tube use. Additional dosimetric monitoring should be performed for each unique irradiation setup.

PET/SPECT/spectral-CT/CBCT imaging in a small-animal radiation therapy platform: A Monte Carlo study-Part I: Quad-modal imaging.

In spite of the tremendous potential of game-changing biological image- and/or biologically guided radiation therapy (RT) and adaptive radiation therapy for cancer treatment, existing limited strategies for integrating molecular imaging and/or biological information with RT have impeded the translation of preclinical research findings to clinical applications. Additionally, there is an urgent need for a highly integrated small-animal radiation therapy (SART) platform that can seamlessly combine therapeutic and diagnostic capabilities to comprehensively enhance RT for cancer treatment. We investigated a highly integrated quad-modal on-board imaging configuration combining positron emission tomography (PET), single-photon emission computed tomography (SPECT), photon-counting spectral CT, and cone-beam computed tomography (CBCT) in a SART platform using a Monte Carlo model as a proof-of-concept. The quad-modal on-board imaging configuration of the SART platform was designed and evaluated by using the GATE Monte Carlo code. A partial-ring on-board PET imaging subsystem, utilizing advanced semiconductor thallium bromide detector technology, was designed to achieve high sensitivity and spatial resolution. On-board SPECT, photon-counting spectral-CT, and CBCT imaging were performed using a single cadmium zinc telluride flat detector panel. The absolute peak sensitivity and scatter fraction of the PET subsystem were estimated by using simulated phantoms described in the NEMA NU-4 standard. The spatial resolution of the PET image of the platform was evaluated by imaging a simulated micro-Derenzo hot-rod phantom. To evaluate the quantitative imaging capability of the system's spectral CT, the Bayesian eigentissue decomposition (ETD) method was utilized to quantitatively decompose the virtual noncontrast (VNC) electron densities and iodine contrast agent fractions in the Kidney1 inserts mixed with the iodine contrast agent within the simulated phantoms. The performance of the proposed quad-model imaging in the platform was validated by imaging a simulated phantom with multiple imaging probes, including an iodine contrast agent and radioisotopes of 18F and 99mTc. The PET subsystem demonstrated an absolute peak sensitivity of 18.5% at the scanner center, with an energy window of 175-560KeV, and a scatter fraction of only 3.5% for the mouse phantom, with a default energy window of 480-540KeV. The spatial resolution of PET on-board imaging exceeded 1.2mm. All imaging probes were identified clearly within the phantom. The PET and SPECT images agreed well with the actual spatial distributions of the tracers within the phantom. Average relative errors on electron density and iodine contrast agent fraction in the Kidney1 inserts were less than 3%. High-quality PET images, SPECT images, spectral-CT images (including iodine contrast agent fraction images and VNC electron density images), and CBCT images of the simulated phantom demonstrated the comprehensive multimodal imaging capability of the system. The results demonstrated the feasibility of the proposed quad-modal imaging configuration in a SART platform. The design incorporates anatomical, molecular, and functional information about tumors, thereby facilitating successful translation of preclinical studies into clinical practices.

Setup and characterisation according to NEMA NU 4 of the phenoPET scanner, a PET system dedicated for plant sciences

Objective. The phenoPET system is a plant dedicated positron emission tomography (PET) scanner consisting of fully digital photo multipliers with lutetium–yttrium oxyorthosilicate crystals and located inside a custom climate chamber. Here, we present the setup of phenoPET, its data processing and image reconstruction together with its performance. Approach. The performance characterization follows the national electrical manufacturers association (NEMA) standard for small animal PET systems with a number of adoptions due to the vertical oriented bore of a PET for plant sciences. In addition temperature stability and spatial resolution with a hot rod phantom are addressed. Main results. The spatial resolution for a 22Na point source at a radial distance of 5 mm to the center of the field-of-view (FOV) is 1.45 mm, 0.82 mm and 1.88 mm with filtered back projection in radial, tangential and axial direction, respectively. A hot rod phantom with 18F gives a spatial resolution of up to 1.6 mm. The peak noise-equivalent count rates are 550 kcps @ 35.08 MBq, 308 kcps @ 33 MBq and 45 kcps @ 40.60 MBq for the mouse, rat and monkey size scatter phantoms, respectively. The scatter fractions for these phantoms are 12.63%, 22.64% and 55.90%. We observe a peak sensitivity of up to 3.6% and a total sensitivity of up to S A,tot = 2.17%. For the NEMA image quality phantom we observe a uniformity of %STD = 4.22% with ordinary Poisson maximum likelihood expectation-maximization with 52 iterations. Here, recovery coefficients of 0.12, 0.64, 0.89, 0.93 and 0.91 for 1 mm, 2 mm, 3 mm, 4 mm and 5 mm rods are obtained and spill-over ratios of 0.08 and 0.14 for the water-filled and air-filled inserts, respectively. Significance. The phenoPET and its laboratory are now in routine operation for the administration of [11C]CO2 and non-invasive measurement of transport and allocation of 11C-labelled photoassimilates in plants.

A Technique to Quantify Very Low Activities in Regions of Interest With a Collimatorless Detector.

We present a new method to measure sub-microcurie activities of photon-emitting radionuclides in organs and lesions of small animals in vivo. Our technique, named the collimator-less likelihood fit, combines a very high sensitivity collimatorless detector with a Monte Carlo-based likelihood fit in order to estimate the activities in previously segmented regions of interest along with their uncertainties. This is done directly from the photon projections in our collimatorless detector and from the region of interest segmentation provided by an x-ray computed tomography scan. We have extensively validated our approach with 225Ac experimentally in spherical phantoms and mouse phantoms, and also numerically with simulations of a realistic mouse anatomy. Our method yields statistically unbiased results with uncertainties smaller than 20% for activities as low as ~111Bq (3nCi) and for exposures under 30 minutes. We demonstrate that our method yields more robust recovery coefficients when compared to SPECT imaging with a commercial pre-clinical scanner, specially at very low activities. Thus, our technique is complementary to traditional SPECT/CT imaging since it provides a more accurate and precise organ and tumor dosimetry, with a more limited spatial information. Finally, our technique is specially significant in extremely low-activity scenarios when SPECT/CT imaging is simply not viable.

Anatomically Realistic 3D Printed Mouse Phantom for Multi-Institutional Benchmarking of FLASH and CONV Irradiation

It is reported that about US$28B/year is spent on pre-clinical studies that are not reproducible. FLASH studies may suffer from the same reproducibility crisis due to the non-standard nature of the FLASH beamlines and the lack of dosimeters that can function at ultra-high dose-rates. There have been reports of different outcomes with regard to the FLASH effect across different institutions, even though similar beamlines, temporal structure, and nominal dose levels were used. This brings up the question of the accuracy of dosimetry under FLASH conditions for a fair comparison between FLASH and CONV. To answer this question, we develop and characterize an anatomically realistic 3D-printed mouse phantom to be used in a multi-institutional dosimetric benchmarking effort. Mesh files for bony anatomy, lungs, and soft tissue derived from a CT scan of a mouse were converted to an editable 3D model. The 3D model was cut along the coronal plane and modified to allow the inclusion of radiographic film. A multi-material approach was employed to print the phantom. A dual-nozzle 3D printer was used, where one of the nozzles used Acrylonitrile butadiene styrene (ABS) to mimic soft tissue and the other nozzle used Polyactic acid (PLA) to mimic bone density. The two materials were used together in a single print. Lungs were approximated by lightweight PLA and were printed separately and inserted into corresponding cavities in the phantom. Hounsfield Units (HU) and print-to-print stability were verified. Radiographic films were laser cut for different anatomical sites. Two institutes took part in this study with data pending from 3 more institutions. The institutes were instructed to deliver 10 Gy to the plane of the film for the whole abdomen, whole lung, and brain irradiations. 2D dose maps were compared between FLASH and CONV, and the deviation from the prescribed dose was also measured. The 3D-printed soft tissue, bone, and lung densities were measured to be ∼ 1.01 g/cc, 1.22 g/cc, and 0.44 g/cc, respectively. For soft tissue and bone, the Hounsfield unit (HU) difference from one print to another was < 10 HU. The greatest variation was within the lungs (54 HU), but this had a minimal effect on the dose distribution (<1%). For the two institutions that completed the survey, the maximum average difference between FLASH and CONV for all irradiations was 0.75 Gy (7.48%). The maximum average difference from the prescribed dose for all irradiations was 0.7 Gy (7.20%) across both institutions. The largest discrepancy was generally observed to be for lung irradiation, indicating that lack of treatment planning systems limits our ability to prescribe accurately in areas of inhomogeneities. A 3D printed anatomically realistic mouse phantom was developed, characterized, and used in a multi-institutional dosimetric benchmarking effort. Such a study is paramount for the clinical translation of FLASH as it facilitates reduced variability from one institution to another.

Technical Infrastructure for Clinical Translation of Electron FLASH

For safe clinical translation of electron FLASH, hardware tools for real-time beam control and software tools for treatment planning are necessary. The purpose of this study is to prototype high-throughput hardware for real-time beam control, along with accurate beam modeling of a modern clinical Linac configured to deliver FLASH dose-rates. For real-time beam current monitoring, a beam current transformer (BCT) was initially coupled to a fast digitizer and its linearity was established by varying dose per pulse. The radiation pulse width was modified, and this change was measured using the BCT. The BCT was then used to measure the variability of dose per pulse and pulse width due to a mistuned linear accelerator system. Next, the BCT was interfaced with a field programmable gate array (FPGA) which provides the ability for high-throughput and deterministic control of the Linac based on dose accumulation. For beam modeling, the program, TOol for PArticle Simulation (TOPAS), was used to obtain beam parameters by using Bayesian optimization of the beam energy, source size, angular, and energy spread via comparison of simulated and representative dose profiles. The beam model would then be employed to calculate 3D dose distribution in a CT scan of a 3D-printed anatomically realistic mouse phantom. The area under the current-time curve from the BCT exhibited excellent linearity (response = 12.80 nC/Gy) up to 2.5 Gy/Pulse (R2 = 0.99). The peak beam current for the electron FLASH beam was measured to be ∼10 mA for an instantaneous dose-rate of ∼5×105 Gy/s. The measured radiation pulse width agreed with the expected value (3.7 μs). The pulse width was then shortened and the measurement by the BCT indicated pulse widths of 1.8 μs and 0.5 μs corresponding to 0.7 Gy/pulse and 0.3 Gy/pulse, respectively. The beamline exhibited a ramp-up in dose per pulse and pulse width when using the automatic frequency controller (AFC). For the first pulse, the dose delivered was ∼0.1-0.3 Gy and the pulse width was 0.6 μs. The output stabilized to nominal values of dose and pulse width after 3-4 pulses. This ramp-up was mitigated by manually tuning the RF resonance with the AFC disabled, after which the BCT exhibited constant output and pulse width. The beam modeling work is in progress. We demonstrated that a BCT can provide real-time measurement of per-pulse output suitable as input for FLASH beam control based on dose accumulation. The next steps are to quantify the accuracy of the dose control mechanism with the FPGA-based hardware. Potential failure modes will be identified and mitigated in parallel with the development of the hardware. A 3D-printed mouse phantom has been constructed to facilitate beam modeling work for treatment planning (in progress). On completion of this work, it is expected that we will have key infrastructure elements needed to move towards an eventual FDA investigational device exemption for clinical trials.

Commissioning and Assessment of Radiation Field and Dose Inhomogeneity for a Dual X-Ray Tube Cabinet Irradiator: To Ensure Accurate Dosimetry in Radiation Biology Experiments

Standardization of X-ray cabinet irradiators calibration and dose reporting is an ongoing process. Multi-tube designs have been introduced and provide theoretical benefit but have not been assessed for pre-clinical irradiation conditions. This study was to report our experience commissioning a dual x-ray tube cabinet irradiator and assess dose distribution for various conditions. Homogeneity and percent depth dose (PDD) of various tube designs in single-tube setting have been previously reported, however multi-tube designs can overcome field asymmetry caused by the heel effect and improve dose fall-off at depth. Profile measurements and output calibration were performed with a calibrated ion chamber. We measured the percent-depth-dose (PDD) with film for each tube individually (PDDind) and both tubes simultaneously (PDDsim). To replicate conditions similar to petri dishes, we evaluated the range of attenuation and scatter under variable solid water thickness up to 9 mm with 3 mm build-up below (i.e., 1.2 cm total backscatter) under several geometries. The films were placed at a fixed position with variable amount of solid water placed above the film. We measured the output with variable water volumes with a 6 cm petri dish to simulate variable solution volumes that are frequently used during in vitro experiments. Film was also placed in a 3D printed mouse phantom with dimensions of (9.5 × 3.6 × 3.0 cm) and constructed to be roughly cylindrical. The dose in the 20 × 20 cm2 tray was within 8.4% of the central axis dose with both tubes. Dose rates decreased at the furthest points on the tray, but remained within 5% within the 15 × 15 cm2 central area. PDDsim with total 7.9 cm backscatter shows that the output is within 95% of the maximum dose at 4 cm depth. PDDsim with 1.2 cm backscatter is also within 95% of the maximum. The PDDind was less than 90% at 1.8 cm, which is similar to other single-tube systems. Slight increase in dose was seen with increasing solution; however, the doses were within 7% of the measured max dose at 6.3 mm depth. This agrees well with the solid water measurements. The results for both solid water and petri dish with variable medium is shown in Table 1. The 3D printed mouse phantom profiles for each plane showed results consistent with the ion chamber measurements. X-ray cell culture irradiation with dual-tube cabinet irradiation is robust when utilizing established dosimetric tools to confirm output and homogeneity. The conditions assumed for calibrations are often not maintained during experiments. We have confirmed that inhomogeneities are present for single tube use, however they are reduced with simultaneous tube use. Additional monitoring should be performed for each unique irradiation setup.

Dosimetry of the PIM1 Pion Beam at the Paul Scherrer Institute for Radiobiological Studies of Mice.

Significant past work has identified unexpected risks of central nervous system (CNS) exposure to the space radiation environment, where long-lasting functional decrements have been associated with multiple ion species delivered at low doses and dose rates. As shielding is the only established intervention capable of limiting exposure to the dangerous radiation fields in space, the recent discovery that pions, emanating from regions of enhanced shielding, can contribute significantly to the total absorbed dose on a deep space mission poses additional concerns. As a prerequisite to biological studies evaluating pion dose equivalents for various CNS exposure scenarios of mice, a careful dosimetric validation study is required. Within our ultimate goal of evaluating the functional consequences of defined pion exposures to CNS functionality, we report in this article the detailed dosimetry of the PiMI pion beam line at the Paul Scherrer Institute, which was developed in support of radiobiological experiments. Beam profiles and contamination of the beam by protons, electrons, positrons and muons were characterized prior to the mice irradiations. The dose to the back and top of the mice was measured using thermoluminescent dosimeters (TLD) and optically simulated luminescence (OSL) to cross-validate the dosimetry results. Geant4 Monte Carlo simulations of radiation exposure of a mouse phantom in water by charged pions were also performed to quantify the difference between the absorbed dose from the OSL and TLD and the absorbed dose to water, using a simple model of the mouse brain. The absorbed dose measured by the OSL dosimeters and TLDs agreed within 5-10%. A 30% difference between the measured absorbed dose and the dose calculated by Geant4 in the dosimeters was obtained, probably due to the approximated Monte Carlo configuration compared to the experiment. A difference of 15-20% between the calculated absorbed dose to water at a 5 mm depth and in the passive dosimeters was obtained, suggesting the need for a correction factor of the measured dose to obtain the absorbed dose in the mouse brain. Finally, based on the comparison of the experimental data and the Monte Carlo calculations, we consider the dose measurement to be accurate to within 15-20%.

Characterization of a commercial bioluminescence tomography-guided system for pre-clinical radiation research.

Widely used Cone-beam computed tomography (CBCT)-guided irradiators have limitations in localizing soft tissue targets growing in a low-contrast environment. This hinders small animal irradiators achieving precise focal irradiation. To advance image-guidance for soft tissue targeting, we developed a commercial-grade bioluminescence tomography-guided system (BLT, MuriGlo) for pre-clinical radiation research. We characterized the system performance and demonstrated its capability in target localization. We expect this study can provide a comprehensive guideline for the community in utilizing the BLT system for radiation studies. MuriGlo consists of four mirrors, filters, lens, and charge-coupled device (CCD) camera, enabling a compact imaging platform and multi-projection and multi-spectral BLT. A newly developed mouse bed allows animals imaged in MuriGlo and transferred to a small animal radiation research platform (SARRP) for CBCT imaging and BLT-guided irradiation. Methods and tools were developed to evaluate the CCD response linearity, minimal detectable signal, focusing, spatial resolution, distortion, and uniformity. A transparent polycarbonate plate covering the middle of the mouse bed was used to support and image animals from underneath the bed. We investigated its effect on 2D Bioluminescence images and 3D BLT reconstruction accuracy, and studied its dosimetric impact along with the rest of mouse bed. A method based on pinhole camera model was developed to map multi-projection bioluminescence images to the object surface generated from CBCT image. The mapped bioluminescence images were used as the input data for the optical reconstruction. To account for free space light propagation from object surface to optical detector, a spectral derivative (SD) method was implemented for BLT reconstruction. We assessed the use of the SD data (ratio imaging of adjacent wavelength) in mitigating out of focusing and non-uniformity seen in the images. A mouse phantom was used to validate the data mapping. The phantom and an in vivo glioblastoma model were utilized to demonstrate the accuracy of the BLT target localization. The CCD response shows good linearity with<0.6% residual from a linear fit. The minimal detectable level is 972 counts for 10×10 binning. The focal plane position is within the range of 13-18mm above the mouse bed. The spatial resolution of 2D optical imaging is<0.3mm at Rayleigh criterion. Within the region of interest, the image uniformity is within 5% variation, and image shift due to distortion is within 0.3mm. The transparent plate caused<6% light attenuation. The use of the SD imaging data can effectively mitigate out of focusing, image non-uniformity, and the plate attenuation, to support accurate multi-spectral BLT reconstruction. There is<0.5% attenuation on dose delivery caused by the bed. The accuracy of data mapping from the 2D bioluminescence images to CBCT image is within 0.7mm. Our phantom test shows the BLT system can localize a bioluminescent target within 1mm with an optimal threshold and only 0.2mm deviation was observed for the case with and without a transparent plate. The same localization accuracy can be maintained for the in vivo GBM model. This work is the first systematic study in characterizing the commercial BLT-guided system. The information and methods developed will be useful for the community to utilize the imaging system for image-guided radiation research.

Development of a morphologically realistic mouse phantom for pre-clinical photoacoustic imaging.

Characterizations based on anatomically realistic phantoms are highly effective to perform accurate technical validation of imaging systems. Specifically for photoacoustic imaging (PAI), although a variety of phantom models with simplified geometries are reported, an unmet need still exists to establish morphologically realistic heterogeneous pre-clinical phantoms. So the development of a mouse-mimicking phantom can reduce the use of animals for the validation and standardization studies of pre-clinical PAI systems and thus eventually translate the PAI technology to clinical research. Here we designed, developed, and fabricated a stable phantom that mimics the detailed morphology of a mouse, to be used as a realistic tool for PAI. The mouse phantom, has been designed by using a combination of image modeling and 3D-printing techniques. As a tissue-mimicking material, we have used copolymer-in-oil-based material that was recently proposed by the International Photoacoustic Standardization Consortium (IPASC). In particular, the anatomically realistic phantom has been modeled by using the real atlas of a mouse as a reference. The mouse phantom includes a 3D-printed skeleton and the main abdominal organs such as the liver, spleen, and kidneys obtained by using doped copolymer-in-oil material with 3D-printed molds. In addition, the acoustic and optical properties of the tissue-mimicking material and the long-term stability have been broadly characterized. Furthermore, our studies showed that thephantom is durable and stable for more than 200 days, under normal storage and repeated use. Fabrication protocol is easy to reproduce. As a result, the proposed morphologically realistic mouse phantom offers durability, material compatibility, and an unprecedented realistic resemblance to the actual rodents' anatomy in PAI. This durable morphologically realistic mouse phantom would minimize the animal experiments in compliance with the 3R principle of Replacement, Reduction, and Refinement. To our knowledge, this is the first time an anatomically realistic heterogeneous mouse phantom has been proposed for PAI in pre-clinical animal imaging and tested its durability over 200 days.