SignificanceThe arterial input function (AIF) plays a crucial role in correcting the time-dependent concentration of the contrast agent within the arterial system, accounting for variations in agent injection parameters (speed, timing, etc.) across patients. Understanding the significance of the AIF can enhance the accuracy of tissue vascular perfusion assessment through indocyanine green–based dynamic contrast-enhanced fluorescence imaging (DCE-FI).AimWe evaluate the impact of the AIF on perfusion assessment through DCE-FI.ApproachA total of 144 AIFs were acquired from 110 patients using a pulse dye densitometer. Simulation and patient intraoperative imaging were conducted to validate the significance of AIF for perfusion assessment based on kinetic parameters extracted from fluorescence images before and after AIF correction. The kinetic model accuracy was evaluated by assessing the variability of kinetic parameters using individual AIF versus population-based AIF.ResultsIndividual AIF can reduce the variability in kinetic parameters, and population-based AIF can potentially replace individual AIF for estimating wash-out rate (kep), maximum intensity (Imax), ingress slope with lower differences compared with those in estimating blood flow, volume transfer constant (Ktrans), and time to peak.ConclusionsIndividual AIF can provide the most accurate perfusion assessment compared with assessment without AIF or based on population-based AIF correction.
SignificanceTissue phantoms that mimic the optical and radiologic properties of human or animal tissue play an important role in the development, characterization, and evaluation of imaging systems. Phantoms that are easily produced and stable for longitudinal studies are highly desirable.AimA new type of long-lasting phantom was developed with commercially available materials and was assessed for fabrication ease, stability, and optical property control. Magnetic resonance imaging (MRI) and x-ray computed tomography (CT) contrast properties were also evaluated.ApproachA systematic investigation of relationships between concentrations of skin-like pigments and composite optical properties was conducted to realize optical property phantoms in the red and near-infrared (NIR) wavelength range that also offered contrast for CT and MRI.ResultsPhantom fabrication time was <1 h and did not involve any heating or cooling processes. Changes in optical properties were <2 % over a 12-month period. Phantom optical and spectral features were similar to human soft tissue over the red to NIR wavelength ranges. Pigments used in the study also had CT and MRI contrasts for multimodality imaging studies.ConclusionsThe phantoms described here mimic optical properties of soft tissue and are suitable for multimodality imaging studies involving CT or MRI without adding secondary contrast agents.
Indocyanine green (ICG)-based dynamic contrast-enhanced fluorescence imaging (DCE-FI) can objectively assess bone perfusion intraoperatively. However, it is susceptible to motion artifacts due to patient’s involuntary respiration during the 4.5-minute DCE-FI data acquisition. An automated motion correction approach based on mutual information (MI) frame-by-frame was developed to overcome this problem. In this approach, MIs were calculated between the reference and the adjacent frame translated and the maximal MI corresponded to the optimal translation. The images obtained from eighteen amputation cases were utilized to validate the approach and the results show that this correction can significantly reduce the motion artifacts and can improve the accuracy of bone perfusion assessment.
KEYWORDS: Magnetic resonance imaging, Imaging systems, Interfaces, Signal detection, Data acquisition, Breast cancer, Tomography, Imaging spectroscopy, Multimodal imaging, Near infrared spectroscopy
A novel MRI-guided near-infrared spectroscopic tomographic imaging system (NIRST) has been developed for breast cancer detection. NIRST imaging for an entire breast can be simultaneously carried out during MRI scanning in less than 4 minutes. Reconstructed phantom images showed clear contrast of a 20 mm inclusion to the background, and the total hemoglobin (HbT) and water concentration values estimated from the reconstructed images of a normal subject were in the same range as those obtained in our previous imaging studies.
ICG-based dynamic contrast-enhanced fluorescence imaging (DCE-FI) and intraoperative DCE- magnetic resonance imaging (MRI) have been carried out nearly simultaneously in three lower extremity bone infection cases to investigate the relationship between these two imaging modalities for assessing bone blood perfusion during open orthopedic surgeries. Time-intensity curves in the corresponding regions of interest of two modalities were derived for comparison. The results demonstrated that ICG-based DCE-FI has higher sensitivity to perfusion changes while DCE-MRI provides superior and supplemental depth-related perfusion information. Research applying the depth-related perfusion information derived from MRI to improve the overall analytic modeling of intraoperative DCE-FI is ongoing.
KEYWORDS: Breast cancer, Multimodal imaging, Tomography, Magnetic resonance imaging, Optical fibers, Data acquisition, Imaging systems, Interfaces, Near infrared spectroscopy
A novel wearable MRg-NIRST system for breast cancer detection has been designed and developed. In this prototype, eight (8) flex circuit strips, each with six (6) photo-detectors (PDs) and six (6) source fibers, are attached to the breast to collect diffused light. A 6x48 fiber switch and 48 side-firing fibers deliver intensity modulated laser light at six (6) near-infrared wavelengths. Light intensity at each of 2304 source-detector positionsis obtained for T2-MRI guided 3D NIRST image reconstruction. In phantom testing, reconstructed images showed the contrast between tumor/inclusion and normal/background.
Significance: The Cherenkov emission spectrum overlaps with that of ambient room light sources. Choice of room lighting devices dramatically affects the efficient detection of Cherenkov emission during patient treatment.
Aim: To determine optimal room light sources allowing Cherenkov emission imaging in normally lit radiotherapy treatment delivery rooms.
Approach: A variety of commercial light sources and long-pass (LP) filters were surveyed for spectral band separation from the red to near-infrared Cherenkov light emitted by tissue. Their effects on signal-to-noise ratio (SNR), Cherenkov to background signal ratio, and image artifacts were quantified by imaging irradiated tissue equivalent phantoms with an intensified time-gated CMOS camera.
Results: Because Cherenkov emission from tissue lies largely in the near-infrared spectrum, a controlled choice of ambient light that avoids this spectral band is ideal, along with a camera that is maximally sensitive to it. An RGB LED light source produced the best SNR out of all sources that mimic room light temperature. A 675-nm LP filter on the camera input further reduced ambient light detected (optical density > 3), achieving maximal SNR for Cherenkov emission near 40. Reduction of the room light signal reduced artifacts from specular reflection on the tissue surface and also minimized spurious Cherenkov signals from non-tissue features such as bolus.
Conclusions: LP filtering during image acquisition for near-infrared light in tandem with narrow band LED illuminated rooms improves image quality, trading off the loss of red wavelengths for better removal of room light in the image. This spectral filtering is also critically important to remove specular reflection in the images and allow for imaging of Cherenkov emission through clear bolus. Beyond time-gated external beam therapy systems, the spectral separation methods can be utilized for background removal for continuous treatment delivery methods including proton pencil beam scanning systems and brachytherapy.
Forty two patients with high energy open fractures were involved into the study to investigate whether an indocyanine green (ICG)-based dynamic contrast-enhanced fluorescence imaging (DCE-FI) can be used to objectively assess bone perfusion and guide surgical debridement. For each patient, fluorescence images were recorded after 0.1 mg/kg of ICG was administered intravenously. By utilizing a bone-specific kinetic model to the video sequences, the perfusion-related metrics were calculated. The results of this study shown that the quantitative ICG-based DEC-FI can accurately assess the human bone perfusion during the orthopedic surgery.
A new low-cost imaging system has been developed for MRI-guided Near-Infrared Spectral Tomography (MRI-NIRST) for breast cancer detection. In this new system, 8 flexible sensing strips with 4 source fibers and 4 photodetectors will be applied directly to the breast, from nipple towards chest-wall, to cover the entire breast. For each source illumination, data will be collected at 32 detector locations. The feasibility of this new imaging system was demonstrated using 3D simulation breast phantoms that created from real breast MR images. Our results indicated that total hemoglobin in a tumor smaller than 1 mm could be recovered with this system.
KEYWORDS: Breast, Magnetic resonance imaging, Tomography, Spectroscopy, Near infrared spectroscopy, Near infrared, Interfaces, Imaging spectroscopy, Light, Imaging systems
A new photo-detector (PD) and source fiber based wearable MRI-guide near infrared spectroscopic tomographic imaging (MRg-NIRST) system has been designed and is underdevelopment. In this new system, eight flex circuit strips, each with 6 PDs and 6 side-firing fibers will be applied to the breast, prior to coil/biopsy positioning. The intensity of the diffused light will be acquired at 48x68 source-detector positions at 12 wavelengths. The 3D optical image reconstruction will be guided through the structural information obtained from T2 MRI and the scattering properties associated with MRI defined breast density.
Significance: Deep-tissue penetration by x-rays to induce optical responses of specific molecular reporters is a new way to sense and image features of tissue function in vivo. Advances in this field are emerging, as biocompatible probes are invented along with innovations in how to optimally utilize x-ray sources.
Aim: A comprehensive review is provided of the many tools and techniques developed for x-ray-induced optical molecular sensing, covering topics ranging from foundations of x-ray fluorescence imaging and x-ray tomography to the adaptation of these methods for sensing and imaging in vivo.
Approach: The ways in which x-rays can interact with molecules and lead to their optical luminescence are reviewed, including temporal methods based on gated acquisition and multipoint scanning for improved lateral or axial resolution.
Results: While some known probes can generate light upon x-ray scintillation, there has been an emergent recognition that excitation of molecular probes by x-ray-induced Cherenkov light is also possible. Emission of Cherenkov radiation requires a threshold energy of x-rays in the high kV or MV range, but has the advantage of being able to excite a broad range of optical molecular probes. In comparison, most scintillating agents are more readily activated by lower keV x-ray energies but are composed of crystalline inorganic constituents, although some organic biocompatible agents have been designed as well. Methods to create high-resolution structured x-ray-optical images are now available, based upon unique scanning approaches and/or a priori knowledge of the scanned x-ray beam geometry. Further improvements in spatial resolution can be achieved by careful system design and algorithm optimization. Current applications of these hybrid x-ray-optical approaches include imaging of tissue oxygenation and pH as well as of certain fluorescent proteins.
Conclusions: Discovery of x-ray-excited reporters combined with optimized x-ray scan sequences can improve imaging resolution and sensitivity.
Significance: The necessity to use exogenous probes for optical oxygen measurements in radiotherapy poses challenges for clinical applications. Options for implantable probe biotechnology need to be improved to alleviate toxicity concerns in human use and facilitate translation to clinical trial use.
Aim: To develop an implantable oxygen sensor containing a phosphorescent oxygen probe such that the overall administered dose of the probe would be below the Federal Drug Administration (FDA)-prescribed microdose level, and the sensor would provide local high-intensity signal for longitudinal measurements of tissue pO2.
Approach: PtG4, an oxygen quenched dendritic molecule, was mixed into an agarose matrix at 100 μM concentration, allowing for local injection into tumors at the total dose of 10 nmol per animal, forming a gel at the site of injection. Cherenkov-excited luminescence imaging (CELI) was used to acquire the phosphorescence and provide intratumoral pO2.
Results: Although PtG4 does not form covalent bonds with agarose and gradually leaches out into the surrounding tissue, its retention time within the gel was sufficiently long to demonstrate the capability to measure intratumoral pO2 with the implantable gel sensors. The sensor’s performance was first evaluated in vitro in tissue simulation phantoms, and then the sensor was used to measure changes in oxygen in MDA-MB-231 tumors during hypofractionated radiotherapy.
Conclusions: Our study demonstrates that implantable oxygen sensors in combination with CELI present a promising approach for quantifying oxygen changes during the course of radiation therapy and thus for evaluating the tumor response to radiation. By improving the design of the gel–probe composition in order to prevent leaching of the probe into the tissue, biosensors can be created that should allow longitudinal oxygen measurements in tumors by means of CELI while using FDA-compliant microdose levels of the probe and thus lowering toxicity concerns.
Cherenkov excitation of molecular probes in tissue can be used for photodynamic effect or diagnostic luminescence imaging. Excitation of dendritic platinum-based metalloporphyrn molecule complexes as an oxygen sensor (PtG4) has been shown to allow oxygen measurement in vivo with spatial resolution defined by the exciting x-ray beam. By sweeping the beamlet shapes and geometries, and detecting the emitted luminscence lifetime, a highly accurate pO2 distribution can be reconstructed of the tissue volume. This has been shown in tumors as well as in normal tissues, and now for implanted microtubes. Additional imaging of luminescent agents based upon Cherenkov or direct radio-excitation could be used for molecular sensing. The combination of depth, concentration, and radiation dose have been analyzed for the tradeoffs in signal to noise, and the signal to background optimization has been completed. The use of this approach can be applied to any clinical radiation therapy linac.
Luminescence molecular tomography with Cherenkov excitation offers the ability to non-invasively image and quantify temporal changes in fluorescence throughout the body, and then further realize tumor localization. This can be done in radiotherapy to determine the response to treatment in fractionated therapy. To obtain high signal-to-background or signal-to-noise ratio measurement, it is critical to know the best post time point of in-vivo agent-based molecular imaging, which could account on a high signal ratio of target to skin (TSR). For this purpose, ex-vivo murine experiments were performed to quantify the biokinetics and biodistribution of the major organs, plasma, tumor, and skin.
The observed behavior of short-wave infrared (SWIR) light in tissue, characterized by relatively low scatter and subdiffuse photon transport, has generated considerable interest for the potential of SWIR imaging to produce high-resolution, subsurface images of fluorescence activity in vivo. These properties have important implications for fluorescence-guided surgery and preclinical biomedical research. Until recently, translational efforts have been impeded by the conventional understanding that fluorescence molecular imaging in the SWIR regime requires custom molecular probes that do not yet have proven safety profiles in humans. However, recent studies have shown that two readily available near-infrared (NIR-I) fluorophores produce measurable SWIR fluorescence, implying that other conventional fluorophores produce detectable fluorescence in the SWIR window. Using SWIR spectroscopy and wide-field SWIR imaging with tissue-simulating phantoms, we characterize and compare the SWIR emission properties of eight commercially available red/NIR-I fluorophores commonly used in preclinical and clinical research, in addition to a SWIR-specific fluorophore. All fluorophores produce measurable fluorescence emission in the SWIR, including shorter wavelength dyes such as Alexa Fluor 633 and methylene blue. This study is the first to report SWIR fluorescence from six of the eight conventional fluorophores and establishes an important comparative reference for developing and evaluating SWIR imaging strategies for biomedical applications.
Cherenkov-excited luminescence scanned imaging (CELSI) is achieved with External Beam Radiotherapy, to map out molecular luminescence intensity or lifetime in tissue. In order to realize a deeper imaging depth with a reasonable spatial resolution, we optimized the original scanning gesture to do in a similar way to computed tomography (CT) and the image reconstruction was instead used a customized Maximum-likelihood expectation maximization (ML-EM) for CELSI. In tomographic CELSI (TCELSI), tomographic images are generated by irradiating the subject using a sequence of programmed X-ray beams at a fixed projection angle, while sensitive measurement is to take a sum for all image pixels from an intensified charge-coupled device. By restricting the X-ray excitation to a single, narrow beam of radiation, the origin of the optical photons can be inferred regardless of where these photons were detected, and how many times they scattered in tissue. Measurement geometry was designed for clinical expectation: CT scanning was achieved by a clinical linear accelerator (LINAC), where X-ray beam sequence and multiple projections were realized with multi leaf collimator (MLC) and gantry movement, respectively. Furthermore, in most modern External Beam Radiotherapy, MLC movement is synchronized with gantry angle to release a uniform radiation, and some of treatment plans, e.g., Intensity Modulated Radiation Therapy (IMRT), have a potential to match the scanning way mentioned. By including Cherenkov imaging results, medium surface profile can be additionally acquired, which can be used as boundary reference to do depth correction and co-register with molecular images. Resolution phantom studies showed that a 0.3 mm diameter capillary tube containing 0.01 nM luminescent nanospheres could be recognized at a depth of 21 mm into tissue-like media. Small animal imaging with a 1 mm diameter cylindrical target demonstrated that fast 3D data acquisition was achieved by a multi-pinhole collimator to image local luminescence 20mm deep.
Solid tumors often exhibit abnormal morphology which can be characterized by increased permeability and low perfusion. The resulting tumor hypoxia has been correlated with poor prognosis, which may be due to ineffective therapy or survival of more aggressive phenotypes. External beam radiation therapy (EBRT) is often used to treat such tumors, where radiation dose is delivered on a daily fractionated basis over the course of weeks. A non-contact optical method for measuring in vivo oxygen levels during EBRT treatments has been developed to provide early indications of hypoxic tumor environments. This method uses a time-gated intensified imaging device to measure both Cherenkov emissions, which are generated in tissue by high energy electrons traveling faster than the phase-velocity of the medium, and Cherenkov-excited luminescence generated by the oxygen-sensitive phosphorescent compound, PtG4. Murine models have shown the ability to discriminate phosphorescence lifetime changes before and after animal sacrifice. Pixel-maps of the estimated pO2 can be generated from this data to show high spatial variability within a region of interest. By further camera optimization, this method can be expanded to show pO2 distributions for other physiological conditions in near real-time. Our imaging method has the unique ability to be integrated within existing clinical applications while providing a wide-field mapping of oxygen saturation, which is currently unavailable with existing point probes.
Cherenkov emission, which is generated during radiation therapy, can be utilized for imaging that is synergistic with radiation therapy. Cherenkov light can be utilized to excite phosphors, which then can be imaged utilizing Cherenkov excited luminescence scanned imaging (CELSI).
Europium chelate microspheres, which exhibit bright luminescence with long luminescent lifetime, were appended with multiple copies of cetuximab. This will allow for selective imaging of EGFR overexpressing tumors during the course of radiation therapy via CELSI. We have characterized the functionality of the cetuximab loaded microspheres in vitro via ELISA, as well as via fluorescence microscopy in EGFR overexpressing A431 cells. These microspheres were intravenously injected into athymic nude mice bearing A431 flank tumors and allowed to incubate for a series of time points. They were then imaged first via standard fluorescence imaging to determine the ideal time point for visualizing tumors via CELSI. After demonstrating selective accumulation in tumors, imaging was then undertaken in vivo via CELSI. These antibody conjugated europium microspheres provide promise to image tumors selectively with CELSI. Future studies involve conjugating other antibodies to the europium microspheres to utilize in CELSI.
KEYWORDS: Short wave infrared radiation, Luminescence, Cameras, In vivo imaging, Infrared imaging, Infrared radiation, Electron beams, Tissues, Radiotherapy, Absorption
Cherenkov emission induced by external beam radiation therapy from a clinical linear accelerator (LINAC) can be used to excite phosphors deep in biological tissues. As with all luminescence imaging, there is a desire to minimize the spectral overlap between the excitation light and emission wavelengths, here between the Cherenkov and the phosphor. Cherenkov excited short-wavelength infrared (SWIR, 1000 to 1700 nm) fluorescence imaging has been demonstrated for the first time, using long Stokes-shift fluorophore PdSe quantum dots (QD) with nanosecond lifetime and an optimized SWIR detection. The 1 / λ2 intensity spectrum characteristic of Cherenkov emission leads to low overlap of this into the fluorescence spectrum of PdSe QDs in the SWIR range. Additionally, using a SWIR camera itself inherently ignores the stronger Cherenkov emission wavelengths dominant across the visible spectrum. The SWIR luminescence was shown to extend the depth sensitivity of Cherenkov imaging, which could be used for applications in radiotherapy sensing and imaging in human tissue with targeted molecular probes.
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