X-ray luminescence computed tomography (XLCT) is a hybrid molecular imaging modality combining the merits of both X-ray imaging (high spatial resolution) and optical imaging (high sensitivity to tracer nanophosphors). Narrow X-ray beam based XLCT imaging has been demonstrated to have the capacity of high spatial resolution imaging at the cost of the data acquisition time. We have primarily focused on improving the performance of the narrow X-ray beam based XLCT imaging. In a previous study, we proposed a scanning strategy achieved by reducing the scanning step size for improving the spatial resolution from double the X-ray beam size to close to the X-ray beam size. For the imaging speed, we recently introduced a continuous scanning scheme to replace the selective excitation scheme and used a photon counter to replace the oscilloscope to acquire measurement data, yielding a 16 times faster scanning time compared with what used in traditional XLCT systems. In addition, we developed a deep learning based XLCT reconstruction algorithm to reduce the number of projection views in a previous work. Moreover, we previously synthesized and compared biocompatible nanophosphors of distinct X-ray luminescence spectra to make multi-color XLCT imaging possible. Here, based on the previous work, we designed and built a first-of-its-kind fast and three-dimensional XLCT imaging system with the capacity of multi-wavelength measurements. A lab-made image acquisition software has been developed to control the system. We have performed physical experiments and verified the system performance.
X-ray luminescence computed tomography (XLCT) is a hybrid molecular imaging modality combining the merits of both x-ray imaging (high spatial resolution) and optical imaging (high sensitivity to tracer nanophosphors). Narrow x-ray beam based XLCT imaging has shown promise for high spatial resolution imaging, but the slow acquisition speed limits its applications for in vivo imaging. We introduced a continuous scanning scheme to replace the selective excitation scheme to improve imaging speed in a previous study. Under the continuous scanning scheme, the main factor that limits the scanning speed is the data acquisition time at each interval position. In this work, we have used a gated photon counter (SR400, Stanford Research Systems) to replace the high-speed oscilloscope (MDO3104, Tektronix) to acquire measurement data. The gated photon counter only counts the photon peaks in each measurement interval, while the oscilloscope records the entire waveform including both background noise data and photon peak data. The photon counter records much less data without losing any relevant information, which makes it ideal for super-fast three-dimensional (3D) imaging. We have built prototype XLCT imaging systems of both types and performed both single target and multiple target phantom experiments in 3D. The results have verified the feasibility of our proposed photon counter based system and good 3D imaging capabilities of XLCT within a reasonable time, yielding a 14 times faster scanning time compared with the oscilloscope based XLCT system. Now, the total scan time is reduced to 27 seconds per transverse section.
Significance: The ability to detect and localize specific molecules through tissue is important for elucidating the molecular basis of disease and treatment. Unfortunately, most current molecular imaging tools in tissue either lack high spatial resolution (e.g., diffuse optical fluorescence tomography or positron emission tomography) or lack molecular sensitivity (e.g., micro-computed tomography, μCT). X-ray luminescence imaging emerged about 10 years ago to address this issue by combining the molecular sensitivity of optical probes with the high spatial resolution of x-ray imaging through tissue. In particular, x-ray luminescence computed tomography (XLCT) has been demonstrated as a powerful technique for the high-resolution imaging of deeply embedded contrast agents in three dimensions (3D) for small-animal imaging.
Aim: To facilitate the translation of XLCT for small-animal imaging, we have designed and built a small-animal dedicated focused x-ray luminescence tomography (FXLT) scanner with a μCT scanner, synthesized bright and biocompatible nanophosphors as contrast agents, and have developed a deep-learning-based reconstruction algorithm.
Approach: The proposed FXLT imaging system was designed using computer-aided design software and built according to specifications. NaGdF4 nanophosphors doped with europium or terbium were synthesized with a silica shell for increased biocompatibility and functionalized with biotin. A deep-learning-based XLCT image reconstruction was also developed based on the residual neural network as a data synthesis method of projection views from few-view data to enhance the reconstructed image quality.
Results: We have built the FXLT scanner for small-animal imaging based on a rotational gantry. With all major imaging components mounted, the motor controlling the gantry can be used to rotate the system with a high accuracy. The synthesized nanophosphors displayed distinct x-ray luminescence emission, which enables multi-color imaging, and has successfully been bound to streptavidin-coated substrates. Lastly, numerical simulations using the proposed deep-learning-based reconstruction algorithm has demonstrated a clear enhancement in the reconstructed image quality.
Conclusions: The designed FXLT scanner, synthesized nanophosphors, and deep-learning-based reconstruction algorithm show great potential for the high-resolution molecular imaging of small animals.
X-ray fluorescence computed tomography (XFCT) is a molecular imaging technique of x-ray photons, which can be used to sense different elements or nanoparticle (NP) agents inside deep samples or tissues. XFCT has been an active research topic for many years. However, XFCT has not been a popular molecular imaging tool because it has limited molecular sensitivity and spatial resolution. To further investigate XFCT imaging, we present a benchtop XFCT imaging system, in in which a unique pencil beam x-ray source and a ring of x-ray spectrometers were simulated using GATE (Geant4 Application for Tomographic Emission) software. An accelerated majorization minimization (MM) algorithm with an L1 regularization scheme was used to reconstruct the XRF image of Molybdenum (Mo) NP targets from the numerical measurements of GATE simulations. With a low x-ray source output rate, good target localization was achieved with a DICE coefficient of 83.681%. The reconstructed signal intensity of the targets was found to be relatively proportional to the target concentrations if detector number and placement is optimized. The MM algorithm performance was compared with maximum likelihood expectation maximization (ML-EM) and filtered back projection (FBP) algorithms. In the future, the benchtop XFCT imaging system will be tested experimentally.
KEYWORDS: X-ray imaging, Luminescence, Computed tomography, Imaging systems, X-rays, Stereoscopy, 3D image processing, Data acquisition, 3D scanning, 3D acquisition
X-ray luminescence computed tomography (XLCT) imaging is a hybrid molecular imaging modality combining the merits of both conventional x-ray imaging (high spatial resolution) and optical imaging (high measurement sensitivity). The narrow x-ray beam based XLCT imaging has been shown to be promising. However due to the selective excitation scheme, the imaging speed is slow thus limiting its practical applications for in vivo imaging. In this work, we have introduced a continuous scanning scheme to acquire data for each angular projection in one motion, eliminating the previous stepping scheme and reducing the data acquisition time, which makes it feasible for multiple transverse scans for three-dimensional (3D) imaging. We have introduced a high accuracy vertical stage to our focused x-ray beam based XLCT imaging system to perform high-resolution and 3D XLCT imaging. We have also included a scintillator crystal coupled to a PMT to act as a single-pixel detector for boundary detection purposes to replace our previous flat panel x-ray detector. We have verified the feasibility of our proposed scanning scheme and imaging system by performing phantom experimental studies. A phantom was embedded with a set of cylindrical targets with 200 μm edge-to-edge distance and was scanned in our imaging system with the proposed method. To test the feasibility for 3D scanning, we took measurements from 4 transverse slices with a vertical step size of 1 mm. The results of the experiments verified the feasibility of our proposed method to perform 3D XLCT imaging using a narrow x-ray beam in a reasonable time.
High-resolution imaging modalities play a critical role for advancing biomedical sciences. Recently, x-ray luminescence computed tomography (XLCT) imaging was introduced as a hybrid molecular imaging modality that combines the highspatial resolution of x-ray imaging and molecular sensitivity of optical imaging. The narrow x-ray beam based XLCT imaging has been demonstrated to achieve high spatial resolution, even at depth, with great molecular sensitivity. Using a focused x-ray beam as the excitation source, orders of magnitude of increased sensitivity has been verified compared with previous methods with a collimated x-ray beam. In this work, we demonstrate the high-spatial resolution capabilities of our focused x-ray beam based XLCT imaging system by scanning two sets of targets, differing in the target size, embedded inside of two tissue-mimicking cylindrical phantoms. Gd2O2S:Eu3+ targets of 200 μm and 150 μm diameters were created and embedded with the same edge-to-edge distances as their diameters respectively. We scanned and reconstructed a single transverse section and successfully demonstrated that a focused x-ray beam with an average dual-cone size of 125 μm could separate the targets in both phantoms with good shape and location accuracy. We have also improved the current XLCT imaging system to make it feasible for fast three-dimensional XLCT scanning.
X-ray luminescence imaging emerged for about a decade and combines both the high spatial resolution of x-ray imaging with the high measurement sensitivity of optical imaging, which could result in a great molecular imaging tool for small animals. So far, there are two types of x-ray luminescence computed tomography (XLCT) imaging. One uses a pencil beam x-ray for high spatial resolution at a cost of longer measurement time. The other uses cone beam x-ray to cover the whole mouse to obtain XLCT images at a very short time but with a compromised spatial resolution. Here we review these two methods in this paper and highlight the synthesized nanophosphors by different research groups.
We are building a focused x-ray luminescence tomography (FXLT) imaging system, developing a machinelearning based FXLT reconstruction algorithm, and synthesizing nanophosphors with different emission wavelengths. In this paper, we will report our current progress from these three aspects. Briefly, we mount all main components, including the focused x-ray tube, the fiber detector, and the x-ray tube and x-ray detector for a microCT system, on a rotary which is a heavy-duty ring track. A microCT scan will be performed before FXLT scan. For a FXLT scan, we will have four PMTs to measure four fiber detectors at two different wavelengths simultaneously for each linear scan position. We expect the spatial resolution of the FXLT imaging will be around 100 micrometers and a limit of detection of approximately 2 μg/mL (for Gd2O2S:Eu).
X-ray luminescence computed tomography (XLCT) is an emerging hybrid molecular imaging modality and has shown great promises in overcoming the strong optical scattering in deep tissues. Though the narrow x-ray beam based XLCT imaging has been demonstrated to obtain high spatial resolution at depth, it suffers from a relatively long measurement time, hindering its practical applications. Recently, we have designed a focused x-ray beam based XLCT imaging system and have successfully performed imaging in about 7.5 seconds per section for a mouse sized object. However, its high spatial resolution capacity has not been fully implemented yet. In this paper, with a superfine focused x-ray beam we design a focused-x-ray luminescence tomography (FXLT) system for spatial resolution up to 94 μm. First, we have described our design in details. Then, we estimate the performance of the designed FXLT imaging system. Lastly, we have found that the spatial resolution of FXLT can be further improved by reducing the scan step size, which has been demonstrated by numerical simulations.
X-ray luminescence computed tomography (XLCT) is an emerging hybrid imaging modality which has the potential for achieving both high sensitivity and spatial resolution simultaneously. For the narrow x-ray beam-based XLCT imaging, based on previous work, a spatial resolution of about double the x-ray beam size can be achieved using a translate/rotate scanning scheme, taking step sizes equal to the x-ray beam width. To break the current spatial resolution limit, we propose a scanning strategy achieved by reducing the scanning step size to be smaller than the x-ray beam size. We performed four sets of numerical simulations and a phantom experiment using cylindrical phantoms and have demonstrated that our proposed scanning method can greatly improve the XLCT-reconstructed image quality compared with the traditional scanning approach. In our simulations, by using a fixed x-ray beam size of 0.8 mm, we were able to successfully reconstruct six embedded targets as small as 0.5 mm in diameter and with the same edge-to-edge distances by using a scanning step as small as 0.2 mm which is a 1.6 times improvement in the spatial resolution compared with the traditional approach. Lastly, the phantom experiment further demonstrated the efficacy of our proposed method in improving the XLCT image quality, with all image quality metrics improving as the step size decreased.
X-ray luminescence computed tomography (XLCT) is an emerging hybrid molecular imaging modality with great promises in overcoming the strong optical scattering in deep tissues for good spatial resolution. Though the narrow x-ray beam XLCT imaging has been demonstrated to obtain high spatial resolution at depth, it suffers from a relatively long measurement time, hindering its practical applications. Recently, we have designed a focused x-ray beam based XLCT imaging system and have successfully performed imaging in about 12.5 minutes per section imaging for a mouse sized object. Following this previous work, in this current study, we have performed XLCT imaging using our focused x-ray beam for both a tissue-mimicking phantom and for the first time, with a euthanized mouse embedded with a capillary tube target filled with 10.0 mg/mL of GOS:Eu3+ microphosphors and have shown that the data acquisition time could be reduced substantially to less than 10 milliseconds per linear scan step compared to the previous study which used 1 second per linear step. In addition, the targets were reconstructed with a high location accuracy and good shape. In the current setup, the total measurement time for a mouse sized object could be reduced to about 7.5 seconds per section imaging, a major improvement from previous studies.
X-ray luminescence computed tomography (XLCT) is an emerging hybrid molecular imaging modality. In XLCT, high energy x-ray photons excite phosphors emitting optical photons for tomographic image reconstruction. During XLCT, the optical signal obtained is thought to only originate from the embedded phosphor particles. However, numerous studies have reported other sources of optical photons such as in air, water, and tissue that are generated from ionization. These sources of optical photons will provide background noise and will limit the molecular sensitivity of XLCT imaging. In this study, using a water-cooled electron multiplying charge-coupled device (EMCCD) camera, we performed luminescence imaging of water, air, and several tissue mimicking phantoms including one embedded with a target containing 0.01 mg/mL of europium-doped gadolinium oxysulfide (GOS:Eu3+) particles during x-ray irradiation using a focused x-ray beam with energy less than the Cerenkov radiation threshold. In addition, a spectrograph was used to measure the x-ray luminescence spectrum. The phantom embedded with the GOS:Eu3+ target displayed the greatest luminescence intensity, followed by the tissue phantom, and finally the water phantom. Our results indicate that the x-ray luminescence intensity from a background phantom is equivalent to a GOS:Eu3+ concentration of 0.8 μg/mL. We also found a 3-fold difference in the radioluminescence intensity between liquid water and air. From the measurements of the emission spectra, we found that water produced a broad spectrum and that a tissue-mimicking phantom made from Intralipid had a different x-ray emission spectrum than one made with TiO2 and India ink. The measured spectra suggest that it is better to use Intralipid instead if TiO2 as optical scatterer for future XLCT imaging.
Photodynamic therapy (PDT) is a noninvasive phototherapy method that has been clinically approved for many years. During this type of therapy, the photosensitizing agent will be excited by optical photons to generate reactive oxygen species which can kill nearby cancer cells. However, due to the strong optical scattering and absorption of tissue, optical photons can only penetrate tissues in few millimeters which result in the limited applications of PDT to superficial lesions like skin cancers. In this study, to overcome the penetration limitations, we used high-energy photons to excite photosensitizers directly by assuming that high-energy photons generate low-energy optical photons in tissues to excite photosensitizers. Cesium- 137 irradiator has been used as the high-energy photon source. A fiber pigtailed diode laser was used to validate the photosensitizer’s efficacy. We used MPPa as the photosensitizer to treat A549 cancer cell line with different concentrations of drug (10μM/ ml, 5 μM/ml, 2.5 μM/ml, 1 μM/ml and 0 μM/ml). We have performed an irradiation experiment for different time durations of 30 min, 15 min, 7 min to 3 min, respectively, and we also compared different drug concentrations and different exposure durations. Our study not only proved the MPPa PDT method was effective, but also indicated that high-energy photons enhanced PDT could potentially overcome the penetration limitations thus making PDT feasible for deep tissue cancer.
Due to the low x-ray photon utilization efficiency and low measurement sensitivity of the electron multiplying charge coupled device camera setup, the collimator-based narrow beam x-ray luminescence computed tomography (XLCT) usually requires a long measurement time. We, for the first time, report a focused x-ray beam-based XLCT imaging system with measurements by a single optical fiber bundle and a photomultiplier tube (PMT). An x-ray tube with a polycapillary lens was used to generate a focused x-ray beam whose x-ray photon density is 1200 times larger than a collimated x-ray beam. An optical fiber bundle was employed to collect and deliver the emitted photons on the phantom surface to the PMT. The total measurement time was reduced to 12.5 min. For numerical simulations of both single and six fiber bundle cases, we were able to reconstruct six targets successfully. For the phantom experiment, two targets with an edge-to-edge distance of 0.4 mm and a center-to-center distance of 0.8 mm were successfully reconstructed by the measurement setup with a single fiber bundle and a PMT.
X-ray luminescence computed tomography (XLCT) is a hybrid molecular imaging modality that uses high energy x-ray photons to excite nanophosphors (e.g. Europium doped Gadolinium Oxysulfide – GOS: Eu3+) emitting optical photons to be measured by a sensitive detector for image reconstruction. XLCT has potentials to combine both the merits of x-ray imaging (high spatial resolution) and optical imaging (high sensitivity), which makes XLCT an attractive imaging modality to image nanophosphor targets deeply embedded in turbid media. In this study, we have evaluated the sensitivity of XLCT with phantom experiments by scanning targets of different phosphor concentrations at different depths. Cylindrical phantoms embedded with a cylindrical target with varying concentrations of GOS: Eu3+ (27.6 mM, 2.76 mM, 276 μM, and 27.6 μM) were scanned inside our lab made XLCT imaging system for varying scanning depths (6, 11, 16, and 21 mm). We found that XLCT is capable of imaging targets of very low concentrations (27.6 μM or 0.01 mg/mL) at significant depths, such as 21 mm. Our results demonstrate that there is also little variation in the reconstructed target size for different imaging depths for XLCT. We have for the first time, compared the sensitivity of XLCT with that of traditional computed tomography (CT) for phosphor targets. We found that XLCT’s use of x-ray induced photons provides much higher measurement sensitivity and contrast compared to CT which provides image contrast solely based on x-ray attenuation.
Super fine collimated x-ray beam based x-ray luminescence computed tomography (XLCT) has the potential to reconstruct the deeply embedded targets with a spatial resolution of hundreds of micrometers. However, due to the low x-ray photon utilization efficiency and low optical signal sensitivity of the electron multiplying charge coupled device (EMCCD) camera, XLCT usually requires a long measurement time. To overcome this limitation, we propose a fiber based, fast XLCT design, in which optical fiber bundles are applied to collect the emitted optical photons on the phantom surface. Highly sensitive photomultiplier tubes (PMT) with a cooling unit and pre-amplifier are used to measure the photons from the fiber bundles. The PMT outputs are collected by a high-speed data acquisition board. A linear scan is estimated to take about 130 seconds, thus for an XLCT scan with 6 projections, we require 13 minutes for each section, which makes it feasible to have a whole body scan of XLCT. To validate our design, numerical simulations and phantom experiments have been performed. In numerical simulation studies, we have investigated the effect of the number of optical fiber bundle on the XLCT reconstruction. We found that one optical fiber bundle is sufficient to reconstruct the deeply embedded targets if measurements from 6 projections are used. Phantom experiments with multiple targets have been performed to validate the proposed fast XLCT imaging.
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