We describe a substantially improved NIRF-IVUS imaging system to overcome current technical limitations. We have implemented a hybrid rotary joint capable of rotational speeds up to 6,000rpm and developed a NIRF-IVUS imaging catheter with a robust dual-layer drive shaft and a reduced rigid length of 2mm with a catheter size <3.6F. NIRF-IVUS processing software was also improved by implementing a high-speed acquisition trigger and data streaming for fast recording speeds. NIRF-IVUS imaging at speeds of at least 30 fps in phantoms and in vivo arterial disease models will demonstrate the unique capabilities of IVUS-NIRF imaging of plaque pathobiology.
SignificanceIntravascular near-infrared fluorescence (NIRF) imaging aims to improve the inspection of vascular pathology using fluorescent agents with specificity to vascular disease biomarkers. The method has been developed to operate in tandem with an anatomical modality, such as intravascular ultrasound (IVUS), and complements anatomical readings with pathophysiological contrast, enhancing the information obtained from the hybrid examination.AimHowever, attenuation of NIRF signals by blood challenges NIRF quantification. We propose a new method for attenuation correction in NIRF intravascular imaging based on a fluorophore-coated guidewire that is used as a reference for the fluorescence measurement and provides a real-time measurement of blood attenuation during the NIRF examination.ApproachWe examine the performance of the method in a porcine coronary artery ex vivo and phantoms using a 3.2F NIRF-IVUS catheter.ResultsWe demonstrate marked improvement over uncorrected signals of up to 4.5-fold and errors of <11 % for target signals acquired at distances up to 1 mm from the catheter system employed.ConclusionsThe method offers a potential means of improving the accuracy of intravascular NIRF imaging under in vivo conditions.
Micro-optical coherence tomography (µOCT), is an emerging optical imaging approach enabling visualization of tissue microstructures at near cellular level. Small form-factor fiber-optic probes are needed to enable uOCT devices for minimally invasive diagnostic procedures such as coronary catheterization for atherosclerosis evaluation. Manufacturing complexities associated with miniaturizing current fiber-optic probes limit their optical and mechanical performance. We will present details of the design and construction of these miniaturized µOCT probes comprising TPL-based 3D printed optics, along with pre-clinical imaging results from an animal model. This probe is capable of lateral resolution of 5 µm and EDOF exceeding 850 µm in tissue.
We report the use of our multimodal near-infrared fluorescence (NIRF) and OCT imaging system and catheter to perform the first imaging of LUM015 inflammatory activity in rabbit models of atherosclerosis in vivo. Using co-injection and multi-channel intravascular NIRF-OCT, we compared LUM015 (6.2 mg/kg) and preclinical ProSense (VM110, 3.5 mg/kg) fluorescence in the same subject. We found that co-registered fluorescence carpet maps were remarkably similar with a PCC of 0.51 and a Mander’s overlap coefficient of 0.79. Results suggest that LUM015 will be a viable clinical option for intracoronary imaging of plaque inflammatory activity in patients.
Activated platelets play a prominent role in multiple diseases, in particular arterial and venous thrombosis, as well as in atherosclerosis and cancer. More recent studies demonstrate further an important role for platelets in generating a permissive microenvironment for tumor growth, and the promotion of invasion and metastasis. To advance the in vivo study of the biological activity of this activated cell type from basic experimental to a clinical focus, new translatable platelet-specific molecular imaging agents are required. Herein, we report the development of a near-infrared fluorescence probe based upon tirofiban, a clinically-approved small molecule glycoprotein IIb/IIIa inhibitor (GPIIb/IIIa). Through in vitro experiments with human platelets, and in vivo in a murine model of deep vein thrombosis, we demonstrate the avidity of the generated probe for activated platelets, thereby enabling rapid in vivo visualization within the vasculature
KEYWORDS: Optical coherence tomography, Luminescence, Imaging systems, Tissues, Process control, Near infrared, Visualization, Standards development, Lens design, Control systems
We present our next generation clinical dual-modality OCT and near infrared autofluorescence/fluorescence (NIRAF/NIRF) imaging platform. This platform allows combined tissue microstructure visualization (OCT) and obtaining molecular information either by intrinsic tissue near infrared autofluorescence (NIRAF) or by exogenous near infrared fluorescence contrast agents (NIRF). Components of this platform, OCT-NIRAF/NIRF imaging system, rotary junction and catheters, were developed using an industry standard design control processes to enable quality clinical translation. We have identified sources of image degradation in dual-modality catheter-based imaging (e.g. core-cladding crosstalk in OCT, background noise in fluorescence) and present methods to mitigate their effects. We also show catheter fabrication and validation, as well as automated fluorescence sensitivity and distance calibration methods that ensure robust and repeatable system performance.
To extend sensitivity field for effective optoacoustic imaging, a novel concept of a non-mechanical point spread function (PSF) adjustment is proposed. Method was validated on phantoms and showed to be useful for distance-adaptive imaging.
Early detection of high-risk coronary atherosclerosis remains an unmet clinical challenge. We have
previously demonstrated a near-infrared fluorescence catheter system for two-dimensional intravascular
detection of fluorescence molecular probes [1]. In this work we improve the system performance by
introducing a novel high resolution sensor. The main challenge of the intravascular sensor is to provide a
highly focused spot at an application relevant distance on one hand and a highly efficient collection of
emitted light on the other.
We suggest employing a double cladding optical fiber (DCF) in combination with focusing optics to
provide a sensor with both highly focused excitation light and highly efficient fluorescent light collection.
The excitation laser is coupled into the single mode core of DCF and guided through a focusing element
and a right angle prism. The resulting side-fired beam exhibits a small spot diameter (50 μm) throughout a
distance of up to 2 mm from the sensor. This is the distance of interest for intravascular coronary imaging
application, determined by an average human coronary artery diameter. At the blood vessel wall, an
activatable fluorescence molecular probe is excited in the diseased lesions. Next light of slightly shifted
wavelength emits only in the places of the inflammations, associated with dangerous plaques [2]. The
emitted light is collected by the cladding of the DCF, with a large collection angle (NA=0.4). The doublecladding
acts as multimodal fiber and guides the collected light to the photo detection elements. The
sensor automatically rotates and pulled-back, while each scanned point is mapped according to the
amount of detected fluorescent emission. The resulting map of fluorescence activity helps to associate the
atherosclerotic plaques with the inflammation process. The presented detection system is a valuable tool
in the intravascular plaque detection and can help to differentiate the atherosclerotic plaques based on
their biological activity, identify the ones that prone to rupture and therefore require more medical
attention.
KEYWORDS: Luminescence, Intravascular ultrasound, Imaging systems, In vivo imaging, Algorithm development, Signal attenuation, Arteries, Blood, Biology, Near infrared
Intravascular Near-Infrared Fluorescence (NIRF) imaging is a promising imaging modality to image vessel
biology and high-risk plaques in vivo. We have developed a NIRF fiber optic catheter and have presented
the ability to image atherosclerotic plaques in vivo, using appropriate NIR fluorescent probes. Our
catheter consists of a 100/140 μm core/clad diameter housed in polyethylene tubing, emitting NIR laser
light at a 90 degree angle compared to the fiber's axis. The system utilizes a rotational and a
translational motor for true 2D imaging and operates in conjunction with a coaxial intravascular
ultrasound (IVUS) device. IVUS datasets provide 3D images of the internal structure of arteries and are
used in our system for anatomical mapping. Using the IVUS images, we are building an accurate hybrid
fluorescence-IVUS data inversion scheme that takes into account photon propagation through the blood
filled lumen. This hybrid imaging approach can then correct for the non-linear dependence of light
intensity on the distance of the fluorescence region from the fiber tip, leading to quantitative imaging.
The experimental and algorithmic developments will be presented and the effectiveness of the
algorithm showcased with experimental results in both saline and blood-like preparations. The
combined structural and molecular information obtained from these two imaging modalities are
positioned to enable the accurate diagnosis of biologically high-risk atherosclerotic plaques in the
coronary arteries that are responsible for heart attacks.
Laser speckle imaging (LSI) is a novel technique for measuring the mechanical properties of atherosclerotic plaques. In LSI, the decorrelation time constant of speckle intensity fluctuations provides an index of viscoelasticity that is closely related to plaque microstructure and composition. Here, we demonstrate for the first time, the feasibility of conducting LSI in vivo using a prototype 1.5 mm (4.5 Fr) diameter intravascular catheter. Investigation of the catheter performance using human arterial samples ex vivo shows that plaque time constants measured by the LSI catheter correlate well with those measured using a free-space bulk optics system. To demonstrate LSI in vivo, the catheter is interfaced with a portable console for intravascular evaluation in the aorta of a living rabbit. Distinct differences in arterial time constants are identified at normal aortic and stented sites in vivo with intravascular LSI.
Early detection of coronary atherosclerosis is an unmet clinical challenge. The detection system has to be highly
sensitive and possess high spacial resolution, in order to provide precise information of the vulnerable plaque location
and size. Recently molecular fluorescence probes have been identified as efficient inflammation biomarkers for the
inflammation process within vulnerable plaques1 and being used in the proposed application to detect inflamed lesions in
the blood vessel wall.
The general principle of the proposed solution is based on a sensor whose head is guided by an intravascular catheter to
the region of interest (coronary artery). When the sensor illuminates an activated fluorescent probe, located in inflamed
areas of vulnerable plaques, the fluorescence is excited and light is emitted with a slightly shifted spectrum. The emitted
light is being collected by the same sensor head, guided through the optical fiber and finally detected by photo-detectors.
In this way, by detecting emitted fluorescence one can obtain information about the location of vulnerable plaques. The
localization resolution is critically depending on the spot size of the illuminating light beam. Moreover, for a high signal
to noise ratio in the detection electronics, as much fluorescent light as possible has to be collected from the plaque
location.
It has been already demonstrated that using single-mode fibers in combination with graded index fibers, a Gaussian
beam, with adjustable waist position and diameter can be formed, representing the fundamental limit of achievable spot
size2. However, when using single mode fibers in this application, the collection efficiency would be very low due to the
small core diameter of this fiber and thus signal to noise ratio would be strongly reduced.
In this work, we present a solution to this challenge, combining both principles. A single mode fiber in combination with
a graded index fiber is used for illumination purposes, while the fluorescence light is collected by the same fiber, but
employing the cladding/coating total reflection to form a multimode fiber for the backwards propagating light. Thus, a
narrow spot size can be obtained allowing high resolution images, with high signal to noise ratio due to the multimodal
collection scheme. We show preliminary results of spot size and beam diameter measurements from the sensor head and
discuss the implication for the improvement of the current catheter-based detection systems.
KEYWORDS: Luminescence, Molecular imaging, In vivo imaging, Blood, Signal detection, Near infrared, Inflammation, Imaging systems, Arteries, Visualization
New imaging methods are urgently needed to identify high-risk atherosclerotic lesions prior to the onset of myocardial infarction, stroke, and ischemic limbs. Molecular imaging offers a new approach to visualize key biological features that characterize high-risk plaques associated with cardiovascular events. While substantial progress has been realized in clinical molecular imaging of plaques in larger arterial vessels (carotid, aorta, iliac), there remains a compelling, unmet need to develop molecular imaging strategies targeted to high-risk plaques in human coronary arteries. We present recent developments in intravascular near-IR fluorescence catheter-based strategies for in vivo detection of plaque inflammation in coronary-sized arteries. In particular, the biological, light transmission, imaging agent, and engineering principles that underlie a new intravascular near-IR fluorescence sensing method are discussed. Intravascular near-IR fluorescence catheters appear highly translatable to the cardiac catheterization laboratory, and thus may offer a new in vivo method to detect high-risk coronary plaques and to assess novel atherosclerosis biologics.
KEYWORDS: Luminescence, In vivo imaging, Microscopy, Nanoparticles, Laser scanners, Control systems, Arteries, Near infrared, Imaging systems, Reflectivity
Molecular and cellular mechanisms of atherogenesis and its treatment are largely being unraveled by in vitro techniques. We describe methodology to directly image macrophage cell activity in vivo in a murine model of atherosclerosis using laser scanning fluorescence microscopy (LSFM) and a macrophage-targeted, near-infrared fluorescent (NIRF) magnetofluorescent nanoparticle (MFNP). Atherosclerotic apolipoprotein E deficient (apoE -/-) mice (n=10) are injected with MFNP or 0.9% saline, and wild-type mice (n=4) are injected with MFNP as additional controls. After 24 h, common carotid arteries are surgically exposed and prepared for LSFM. Multichannel LSFM of MFNP-enhanced carotid atheroma (5×5-µm in-plane resolution) shows a strong focal NIRF signal, with a plaque target-to-background ratio of 3.9±1.8. Minimal NIRF signal is observed in control mice. Spectrally resolved indocyanine green (ICG) fluorescence angiograms confirm the intravascular location of atheroma. On ex vivo fluorescence reflectance imaging, greater NIRF plaque signal is seen in apoE -/- MFNP mice compared to controls (p<0.01). The NIRF signal correlates well with immunostained macrophages, both by stained surface area (r=0.77) and macrophage number (r=0.86). The validated experimental methodology thus establishes a platform for investigating macrophage activity in atherosclerosis in vivo, and has implications for the detection of clinical vulnerable plaques.
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