Accounting for blood attenuation in intravascular near-infrared fluorescence-ultrasound imaging using a fluorophore-coated guidewire.

Significance: Intravascular 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. Aim: However, 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. Approach: We examine the performance of the method in a porcine coronary artery ex vivo and phantoms using a 3.2F NIRF-IVUS catheter. Results: We 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. Conclusions: The method offers a potential means of improving the accuracy of intravascular NIRF imaging under in vivo conditions.

Intravital Microscopy in Atherosclerosis Research.

Atherosclerosis is a lipid-driven inflammatory disorder that narrows the arterial lumen and can induce life-threatening complications from coronary artery disease, cerebrovascular disease, and peripheral artery disease. On a mechanistic level, the development of novel cellular-resolution intravital microscopy imaging approaches has recently enabled in vivo studies of underlying biological processes governing disease onset and progress. In particular, multiphoton microscopy has emerged as a promising intravital imaging tool utilizing two-photon-excited fluorescence and second-harmonic generation that provides subcellular resolution and increased imaging depths beyond confocal and epifluorescence microscopy. In this chapter, we describe the state-of-the-art multiphoton microscopy applied to the study of murine atherosclerosis.

Intravascular molecular-structural imaging with a miniaturized integrated near-infrared fluorescence and ultrasound catheter.

Coronary artery disease (CAD) remains a leading cause of mortality and warrants new imaging approaches to better guide clinical care. We report on a miniaturized, hybrid intravascular catheter and imaging system for comprehensive coronary artery imaging in vivo. Our catheter exhibits a total diameter of 1.0 mm (3.0 French), equivalent to standalone clinical intravascular ultrasound (IVUS) catheters but enables simultaneous near-infrared fluorescence (NIRF) and IVUS molecular-structural imaging. We demonstrate NIRF-IVUS imaging in vitro in coronary stents using NIR fluorophores, and compare NIRF signal strengths for prism and ball lens sensor designs in both low and high scattering media. Next, in vivo intravascular imaging in pig coronary arteries demonstrates simultaneous, co-registered molecular-structural imaging of experimental CAD inflammation on IVUS and distance-corrected NIRF images. The obtained results suggest substantial potential for the NIRF-IVUS catheter to advance standalone IVUS, and enable comprehensive phenotyping of vascular disease to better assess and treat patients with CAD.

A submicrometre silicon-on-insulator resonator for ultrasound detection.

Ultrasound detectors use high-frequency sound waves to image objects and measure distances, but the resolution of these readings is limited by the physical dimensions of the detecting element. Point-like broadband ultrasound detection can greatly increase the resolution of ultrasonography and optoacoustic (photoacoustic) imaging1,2, but current ultrasound detectors, such as those used for medical imaging, cannot be miniaturized sufficiently. Piezoelectric transducers lose sensitivity quadratically with size reduction3, and optical microring resonators4 and Fabry-Pérot etalons5 cannot adequately confine light to dimensions smaller than about 50 micrometres. Micromachining methods have been used to generate arrays of capacitive6 and piezoelectric7 transducers, but with bandwidths of only a few megahertz and dimensions exceeding 70 micrometres. Here we use the widely available silicon-on-insulator technology to develop a miniaturized ultrasound detector, with a sensing area of only 220 nanometres by 500 nanometres. The silicon-on-insulator-based optical resonator design provides per-area sensitivity that is 1,000 times higher than that of microring resonators and 100,000,000 times better than that of piezoelectric detectors. Our design also enables an ultrawide detection bandwidth, reaching 230 megahertz at -6 decibels. In addition to making the detectors suitable for manufacture in very dense arrays, we show that the submicrometre sensing area enables super-resolution detection and imaging performance. We demonstrate imaging of features 50 times smaller than the wavelength of ultrasound detected. Our detector enables ultra-miniaturization of ultrasound readings, enabling ultrasound imaging at a resolution comparable to that achieved with optical microscopy, and potentially enabling the development of very dense ultrasound arrays on a silicon chip.

Dual-wavelength hybrid optoacoustic-ultrasound biomicroscopy for functional imaging of large-scale cerebral vascular networks.

A critical link exists between pathological changes of cerebral vasculature and diseases affecting brain function. Microscopic techniques have played an indispensable role in the study of neurovascular anatomy and functions. Yet, investigations are often hindered by suboptimal trade-offs between the spatiotemporal resolution, field-of-view (FOV) and type of contrast offered by the existing optical microscopy techniques. We present a hybrid dual-wavelength optoacoustic (OA) biomicroscope capable of rapid transcranial visualization of large-scale cerebral vascular networks. The system offers 3-dimensional views of the morphology and oxygenation status of the cerebral vasculature with single capillary resolution and a FOV exceeding 6 × 8 mm2 , thus covering the entire cortical vasculature in mice. The large-scale OA imaging capacity is complemented by simultaneously acquired pulse-echo ultrasound (US) biomicroscopy scans of the mouse skull. The new approach holds great potential to provide better insights into cerebrovascular function and facilitate efficient studies into neurological and vascular abnormalities of the brain.

Looking at sound: optoacoustics with all-optical ultrasound detection.

Originally developed for diagnostic ultrasound imaging, piezoelectric transducers are the most widespread technology employed in optoacoustic (photoacoustic) signal detection. However, the detection requirements of optoacoustic sensing and imaging differ from those of conventional ultrasonography and lead to specifications not sufficiently addressed by piezoelectric detectors. Consequently, interest has shifted to utilizing entirely optical methods for measuring optoacoustic waves. All-optical sound detectors yield a higher signal-to-noise ratio per unit area than piezoelectric detectors and feature wide detection bandwidths that may be more appropriate for optoacoustic applications, enabling several biomedical or industrial applications. Additionally, optical sensing of sound is less sensitive to electromagnetic noise, making it appropriate for a greater spectrum of environments. In this review, we categorize different methods of optical ultrasound detection and discuss key technology trends geared towards the development of all-optical optoacoustic systems. We also review application areas that are enabled by all-optical sound detectors, including interventional imaging, non-contact measurements, magnetoacoustics, and non-destructive testing.

Quantitative intravascular biological fluorescence-ultrasound imaging of coronary and peripheral arteries in vivo.

AIMS: (i) to evaluate a novel hybrid near-infrared fluorescence-intravascular ultrasound (NIRF-IVUS) system in coronary and peripheral swine arteries in vivo; (ii) to assess simultaneous quantitative biological and morphological aspects of arterial disease. METHODS AND RESULTS: Two 9F/15MHz peripheral and 4.5F/40MHz coronary near-infrared fluorescence (NIRF)-IVUS catheters were engineered to enable accurate co-registrtation of biological and morphological readings simultaneously in vivo. A correction algorithm utilizing IVUS information was developed to account for the distance-related fluorescence attenuation due to through-blood imaging. Corrected NIRF (cNIRF)-IVUS was applied for in vivo imaging of angioplasty-induced vascular injury in swine peripheral arteries and experimental fibrin deposition on coronary artery stents, and of atheroma in a rabbit aorta, revealing feasibility to intravascularly assay plaque structure and inflammation. The addition of ICG-enhanced NIRF assessment improved the detection of angioplasty-induced endothelial damage compared to standalone IVUS. In addition, NIRF detection of coronary stent fibrin by in vivo cNIRF-IVUS imaging illuminated stent pathobiology that was concealed on standalone IVUS. Fluorescence reflectance imaging and microscopy of resected tissues corroborated the in vivo findings. CONCLUSIONS: Integrated cNIRF-IVUS enables simultaneous co-registered through-blood imaging of disease related morphological and biological alterations in coronary and peripheral arteries in vivo. Clinical translation of cNIRF-IVUS may significantly enhance knowledge of arterial pathobiology, leading to improvements in clinical diagnosis and prognosis, and helps to guide the development of new therapeutic approaches for arterial diseases.

Hybrid optical and acoustic resolution optoacoustic endoscopy.

We propose the implementation of hybrid optical and acoustic resolution optoacoustic endoscopy. Laser light is transmitted to tissue by two types of illumination for achieving optical and acoustic resolution imaging. A 20 MHz ultrasound detector is used for recording optoacoustic signals. The endoscopy probe attains a 3.6 mm diameter and is fully encapsulated into a catheter system. We validate the imaging performance of the hybrid endoscope on phantoms and ex vivo, and discuss the necessity for the extended resolution and depth range of endoscopy achieved.

All-optical optoacoustic microscope based on wideband pulse interferometry.

Optical and optoacoustic (photoacoustic) microscopy have been recently joined in hybrid implementations that resolve extended tissue contrast compared to each modality alone. Nevertheless, the application of the hybrid technique is limited by the requirement to combine an optical objective with ultrasound detection collecting signal from the same micro-volume. We present an all-optical optoacoustic microscope based on a pi-phase-shifted fiber Bragg grating (π-FBG) with coherence-restored pulsed interferometry (CRPI) used as the interrogation method. The sensor offers an ultra-small footprint and achieved higher sensitivity over piezoelectric transducers of similar size. We characterize the spectral bandwidth of the ultrasound detector and interrogate the imaging performance on phantoms and tissues. We show the first optoacoustic images of biological specimen recorded with π-FBG sensors. We discuss the potential uses of π-FBG sensors based on CRPI.

Characterization of the spatio-temporal response of optical fiber sensors to incident spherical waves.

In this study a theoretical framework for calculating the acoustic response of optical fiber-based ultrasound sensors is presented. The acoustic response is evaluated for optical fibers with several layers of coating assuming a harmonic point source with arbitrary position and frequency. First, the fiber is acoustically modeled by a layered cylinder on which spherical waves are impinged. The scattering of the acoustic waves is calculated analytically and used to find the normal components of the strains on the fiber axis. Then, a strain-optic model is used to calculate the phase shift experienced by the guided mode in the fiber owing to the induced strains. The framework is showcased for a silica fiber with two layers of coating for frequencies in the megahertz regime, commonly used in medical imaging applications. The theoretical results are compared to experimental data obtained with a sensing element based on a pi-phase-shifted fiber Bragg grating and with photoacoustically generated ultrasonic signals.