KEYWORDS: Breast, Tissue optics, 3D image processing, Signal attenuation, Optoacoustics, Blood, 3D image reconstruction, Blood vessels, Imaging systems, Breast imaging
Significance: In three-dimensional (3D) functional optoacoustic tomography (OAT), wavelength-dependent optical attenuation and nonuniform incident optical fluence limit imaging depth and field of view and can hinder accurate estimation of functional quantities, such as the vascular blood oxygenation. These limitations hinder OAT of large objects, such as a human female breast.
Aim: We aim to develop a measurement-data-driven method for normalization of the optical fluence distribution and to investigate blood vasculature detectability and accuracy for estimating vascular blood oxygenation.
Approach: The proposed method is based on reasonable assumptions regarding breast anatomy and optical properties. The nonuniform incident optical fluence is estimated based on the illumination geometry in the OAT system, and the depth-dependent optical attenuation is approximated using Beer–Lambert law.
Results: Numerical studies demonstrated that the proposed method significantly enhanced blood vessel detectability and improved estimation accuracy of the vascular blood oxygenation from multiwavelength OAT measurements, compared with direct application of spectral linear unmixing without optical fluence compensation. Experimental results showed that the proposed method revealed previously invisible structures in regions deeper than 15 mm and/or near the chest wall.
Conclusions: The proposed method provides a straightforward and computationally inexpensive approximation of wavelength-dependent effective optical attenuation and, thus, enables mitigation of the spectral coloring effect in functional 3D OAT imaging.
The goal of quantitative optoacoustic tomography (qOAT) is to reconstruct a distribution of absolute chromophore concentrations and/or functional properties from measurements of the optically induced pressure (ultrasound signals) acquired at multiple excitation wavelengths. Estimating the distribution of hemoglobin, an endogenous OAT chromophore, is important because the oxygen saturation distribution of the blood vessels is a well-known indicator of aggressive growth of a cancerous tumor. In a number of studies, a spectral linear unmixing method has been applied to two-dimensional slices of tissue acquired with OAT at multiple wavelengths, leading to promising results at moderate penetration depths of ≤ 2 cm. In the three-dimensional (3D) OAT of the breast, such functional images cannot be accurately reconstructed via the spectral linear unmixing method due to unknown spatial distribution of the optical fluence in a relatively large size of the volume of interest (≥ 4 cm). Optical attenuation in biological tissue depends on the optical wavelength, and the optical fluence is exponentially attenuated with increasing imaging depth. Thus, the accuracy of the estimated distribution decreases with depth. To overcome this challenge, we investigated a spectral linear unmixing method with a simplified optical fluence normalization based on measurements of background absorbed optical energy in the breast. We compare estimates of blood oxygen saturations from two-wavelength clinical OAT breast images and demonstrate acceptable accuracy of ~10% while lack of compensation for the optical fluence distribution can lead to values outside the physiological range. We also quantitatively compare the accuracy of oxygen saturation estimates using numerical simulation of photon transport in realistic 3D OAT breast phantoms at dual wavelengths of 757 and 850 nm with inverse ratio of the optical absorption by deoxy- (Hb) and oxy-hemoglobin (HbO2) and three wavelengths of 757, 800, and 850 nm with inclusion of isosbestic point of the optical absorption in Hb/HbO2.
Due to a demonstrated capability to assess tumor angiogenesis and hypoxia in mammalian systems, there is great interest in applying optoacoustic tomography (OAT) to the study and screening of breast cancer. In order to translate OAT to clinical applications, in silico studies are crucial for studying imaging system parameters that might be impossible to assess via direct experimentation. Previous numerical phantoms have proven to be too unrealistic for rigorous testing of modern image reconstruction methods and clinically relevant signal detection tasks. Recently, the U.S. Food and Drug Administration has released software to generate realistic three-dimensional numerical realizations of the human female breast as part of the Virtual Imaging Clinical Trials for Regulatory Evaluation (VICTRE) project. By careful selection of physical attributes and material coefficients, the VICTRE breast phantom can be customized for particular imaging tasks, but no such customization has been given for OAT. We propose a general framework of in silico studies for OAT breast imaging using the VICTRE breast phantom. We will create an ensemble of OAT breast phantoms, using appropriate optical and acoustic parameters, that have typical sizes and tissue densities. Various lesions will be created and embedded based on clinical scenarios. We will define and perform several signal detection tasks by which the system performance may be compared. Generation of such an ensemble requires substantial computation but once produced, it can be utilized in other numerical simulation studies of the configuration of OAT imaging systems customized for diverse tasks. We will make this ensemble of phantoms publicly available online. The proposed framework will permit standardization of the assessment of 3D OAT data-acquisition parameters and image reconstruction methods.
Optoacoustic tomography (OAT) is a promising modality for breast imaging that provides high resolution, detection sensitivity and diagnostic specificity for vascularized breast tumors. In OAT systems employing an arc- shaped illuminator, irregular overlaps of light beams can yield a non-uniform illumination throughout the entire volume of the breast. The imbalance in optical fluence leads to intensity loss in the reconstructed OAT images. Additionally, because optical fluence decreases with depth from breast skin surface, i.e., optical attenuation, deep breast tissues are diminished in the reconstructed images. For qualitative enhancement in 3D OAT imaging, we propose an image processing method to estimate, and compensate for, both the non-uniform incident optical fluence and the optical attenuation. We approximate the non-uniform illumination via maximum intensity extraction for polar angles in a spherical coordinate system. The location of the breast surface is estimated by detecting blood vessels nearest to the breast skin layer that appear with relatively high intensities in the reconstructed image. The breast depth is computed as the minimum distance between each voxel and the detected breast surface. The depth-dependent optical attenuation in the breast is estimated using the Beer– Lambert law down to the maximum penetration depth determined from an analysis of noise and artifacts in the reconstructed image. At each polar angle, the reciprocals of the estimated attenuation is used to compensate for the loss in intensity. The results are that previously invisible structures near the chest wall are revealed, and visible penetration depth was increased by 67% over the conventional, non-compensated volumes.
Laser Optoacoustic Ultrasonic Imaging System Assembly (LOUISA-3D) was developed in response to demand of diagnostic radiologists for an advanced screening system for the breast to improve on low sensitivity of x-ray based modalities of mammography and tomosynthesis in the dense and heterogeneous breast and low specificity magnetic resonance imaging. It is our working hypothesis that co-registration of quantitatively accurate functional images of the breast vasculature and microvasculature, and anatomical images of breast morphological structures will provide a clinically viable solution for the breast cancer care. Functional imaging is LOUISA-3D is enabled by the full view 3D optoacoustic images acquired at two rapidly toggling laser wavelengths in the near-infrared spectral range. 3D images of the breast anatomical background is enabled in LOUISA-3D by a sequence of B-mode ultrasound slices acquired with a transducer array rotating around the breast. This creates the possibility to visualize distributions of the total hemoglobin and blood oxygen saturation within specific morphological structures such as tumor angiogenesis microvasculature and larger vasculature in proximity of the tumor. The system has four major components: (i) a pulsed dual wavelength laser with fiberoptic light delivery system, (ii) an imaging module with two arc shaped probes (optoacoustic and ultrasonic) placed in a transparent bowl that rotates around the breast, (iii) a multichannel electronic system with analog preamplifiers and digital data acquisition boards, and (iv) computer for the system control, data processing and image reconstruction. The most important advancement of this latest system design compared with previously reported systems is the full breast illumination accomplished for each rotational step of the optoacoustic transducer array using fiberoptic illuminator rotating around the breast independently from rotation of the detector probe. We report here a pilot case studies on one healthy volunteer and on patient with a suspicious small lesion in the breast. LOUISA3D visualized deoxygenated veins and oxygenated arteries of a healthy volunteer, indicative of its capability to visualize hypoxic microvasculature in cancerous tumors. A small lesion detected on optoacoustic image of a patient was not visible on ultrasound, potentially indicating high system sensitivity of the optoacoustic subsystem to small but aggressively growing cancerous lesions with high density angiogenesis microvasculature. The main breast vasculature (0.5-1 mm) was visible at depth of up to 40-mm with 0.3-mm resolution. The results of LOUISA-3D pilot clinical validation demonstrated the system readiness for statistically significant clinical feasibility study.
Enhanced delivery of optical clearing agents (OCA) through skin may improve sensitivity of optical and optoacoustic (OA) methods of imaging, sensing, and monitoring. This report describes a two-step method for enhancement of light penetration through skin. Here, we demonstrate that topical application of hyaluronic acid (HA) improves skin penetration of hydrophilic and lipophilic OCA and thus enhances their performance. We examined the OC effect of 100% polyethylene and polypropylene glycols (PPGs) and their mixture after pretreatment by HA, and demonstrated significant increase in efficiency of light penetration through skin. Increased light transmission resulted in a significant increase of OA image contrast in vitro. Topical pretreatment of skin for about 30 min with 0.5% HA in aqueous solution offers effective delivery of low molecular weight OCA such as a mixture of PPG-425 and polyethylene glycol (PEG)-400. The developed approach of pretreatment by HA prior to application of clearing agents (PEG and PPG) resulted in a ∼47-fold increase in transmission of red and near-infrared light and significantly enhanced contrast of OA images.
We describe the ongoing development and performance of a high-pulse-energy wavelength-cycling laser system for three-dimensional optoacoustic tomography of the breast. Joule-level energies are desired for achieving the required penetration depths while maintaining safe fluence levels. Wavelength cycling provides a pulse sequence which repeatedly alternates between two wavelengths (approximately 756 and 797 nm) that provide differential imaging. This improves co-registration of captured differential images and quantification of blood oxygen saturation. New design features have been developed for and incorporated into a clinical prototype laser system, to improve efficacy and ease of use in the clinic. We describe the benefits of these features for operation with a clinical pilot optoacoustic / ultrasound dual-modality three-dimensional imaging system.
We developed a new and improved Laser Optoacoustic Imaging System, LOIS-3D for preclinical research applications in small animal models. The advancements include (i) a new stabilized imaging module with a more homogeneous illumination of the mouse yielding a better spatial resolution (<0.2 mm) and (ii) a new low noise amplifier incorporated into the ultrasonic probe and providing the noise equivalent pressure around 2 Pa resulting in increased signal-to-noise ratio and the optical absorption sensitivity of about 0.15 cm-1. We also improved scan time and the image reconstruction times. This prototype has been commercialized for a number of biomedical research applications, such as imaging vascularization and measuring hemoglobin / oxyhemoglobin distribution in the organs as well as imaging exogenous or endogenous optoacoustic contrast agents. As examples, we present in vivo experiments using phantoms and mice with and without tumor injected with contrast agents with indocyanine green (ICG). LOIS-3D was capable of detecting ~1-2 pmole of the ICG, in tissues with relatively low blood content. With its high sensitivity and excellent spatial resolution LOIS-3D is an advanced alternative to fluorescence and bioluminescence based modalities for molecular imaging in live mice.
In this work, we introduce an improved prototype of the imaging system that combines three-dimensional optoacoustic tomography (3D-OAT) and laser ultrasound tomography slicer (2D-LUT) to obtain coregistered maps of tissue optical absorption and speed of sound (SOS). The imaging scan is performed by a 360 degree rotation of a phantom/mouse with respect to a static arc-shaped array of ultrasonic transducers. A Q-switched laser system is used to establish optoacoustic illumination pattern appropriate for deep tissue imaging with a tunable (730-840 nm) output wavelengths operated at 10 Hz pulse repetition rate. For the LUT slicer scans, the array is pivoted by 90 degrees with respect to the central transducers providing accurate registration of optoacoustic and SOS maps, the latter being reconstructed using waveform inversion with source encoding (WISE) technique. The coregistered OAT-LUT modality is validated by imaging a phantom and a live mouse. SOS maps acquired in the imaging system can be employed by an iterative optoacoustic reconstruction algorithm capable of compensating for acoustic wavefield aberrations. The most promising applications of the imaging system include 3D angiography, cancer research, and longitudinal studies of biological distributions of optoacoustic contrast agents (carbon nanotubes, metal plasmonic nanoparticles, fluorophores, etc.).
Iterative image reconstruction algorithms can model complicated imaging physics, compensate for imperfect data acquisition systems, and exploit prior information regarding the object. Hence, they produce higher quality images than do analytical image reconstruction algorithms. However, three-dimensional (3D) iterative image reconstruction is computationally burdensome, which greatly hinders its use with applications requiring a large field-of-view (FOV), such as breast imaging. In this study, an improved GPU-based implementation of a numerical imaging model and its adjoint have been developed for use with general gradient-based iterative image reconstruction algorithms. Both computer simulations and experimental studies are conducted to investigate the efficiency and accuracy of the proposed implementation for optoacoustic tomography (OAT). The results suggest that the proposed implementation is more than five times faster than the previous implementation.
In this work we introduce an improved prototype of three-dimensional imaging system that combines optoacoustic tomography (OAT) and laser ultrasound tomography (LUT) to obtain coregistered maps of tissue optical absorption and speed of sound (SoS). The OAT scan is performed by a 360 degree rotation of a mouse with respect to an arc-shaped array of ultrasonic transducers. A Q-switched laser system is used to establish optoacoustic illumination pattern appropriate for deep tissue imaging with a tunable (730-840 nm) output wavelengths operated at 10 Hz pulse repetition rate. A 532 nm wavelength output, being mostly absorbed within a narrow superficial layer of skin, is used to outline the visualized biological object. Broadband laser ultrasound emitters are arranged in another arc pattern and are positioned opposite and orthogonal to the array of transducers. This imaging geometry allows reconstruction of volumes that depict SoS distributions from the measured time of flight data. The reconstructed LUT images can subsequently be employed by an optoacoustic reconstruction algorithm to compensate for acoustic wavefield aberration and thereby improve accuracy of the reconstructed images of the absorbed optical energy. The coregistered OAT-LUT imaging is validated in a phantom and live mouse using a single-slice system prototype.
Gold-based contrast agents, gold nanorod (GNR), were designed for the enhancement of optoacoustic signal. After synthesis, the GNR-CTAB complexes were modified by pegylation (PEG), or replacement of CTAB (cetyl trimethylammonium bromide) with MTAB (16-mercaptohexadecyl trimethylammonium bromide) for coverage of gold nanorods with heparin (GNR-HP). Modified GNR are purified through centrifugation and filtration. GNRCTAB can be used as a model of positively charged gold surface for quantitative optoacoustic sensing in GNRbacteria interactions, whereas GNR-PEG and GNR-HP can be used as negatively charged gold surface models. We studied controlled agglomeration of contrast agents with the bacteria E.Coli and Vibrio Cholerae. For bacterial sensing, the localized plasmon resonance peak shifts as a function of electrostatic binding, which was detected with two different wavelengths through 3D optoacoustic imaging.
In this work, we investigate a novel reconstruction method for laser-induced ultrasound computed tomography (USCT) breast imaging that circumvents limitations of existing methods that rely on ray-tracing. There is currently great interest in developing hybrid imaging systems that combine optoacoustic tomography (OAT) and USCT. There are two primary motivations for this: (1) the speed-of-sound (SOS) distribution reconstructed by USCT can provide complementary diagnostic information; and (2) the reconstructed SOS distribution can be incorporated in the OAT reconstruction algorithm to improve OAT image quality. However, image reconstruction in USCT remains challenging. The majority of existing approaches for USCT breast imaging involve ray-tracing to establish the imaging operator. This process is cumbersome and can lead to inaccuracies in the reconstructed SOS images in the presence of multiple ray-paths and/or shadow zones. To circumvent these problems, we implemented a partial differential equation-based Eulerian approach to USCT that was proposed in the mathematics literature but never investigated for medical imaging applications. This method operates by directly inverting the Eikonal equation without ray-tracing. A numerical implementation of this method was developed and compared to existing reconstruction methods for USCT breast imaging. We demonstrated the ability of the new method to reconstruct SOS maps from TOF data obtained by a hybrid OAT/USCT imager built by our team.
A new experimental approach for measurements of temperature dependence of the Grüneisen parameter in optically absorbing solutions is proposed. Two-dimensional optoacoustic (OA) imaging is used to improve accuracy of signal amplitude measurements and spatial localization of the studied samples. We estimated OA response of optically absorbing solutions measuring median intensity of OA images within the region of interest (ROI) as a function of temperature. We showed that when normalized to its value at a particular temperature, OA image intensity becomes an accurate metric reflecting temperature changes of Grüneisen parameter regardless of local optical fluence and absorbance, assuming those remain constant with temperature. Using the proposed method we studied temperature dependence of aqueous solutions of nickel and cupric sulfate in the range from 4 to 40°C. Obtained results were compared with temperature dependence for the Grüneisen parameter of DI-water, which we measured by using carbon ink colloid. We also found that Grüneisen-temperature relationship for nickel sulfate exhibits linear trend with respect to the concentration, and is independent of coupling medium and laser excitation wavelength.In the future, the developed methodology could be adopted for important applications of in vivo optoacoustic temperature monitoring.
KEYWORDS: Ultrasonography, Imaging systems, Acoustics, 3D image processing, Tomography, Visualization, Signal attenuation, 3D modeling, Ultrasound tomography, Pre-clinical research
In this work, we introduce a novel three-dimensional imaging system for in vivo high-resolution anatomical and functional whole-body visualization of small animal models developed for preclinical or other type of biomedical research. The system (LOUIS-3DM) combines a multi-wavelength optoacoustic and ultrawide-band laser ultrasound tomographies to obtain coregistered maps of tissue optical absorption and acoustic properties, displayed within the skin outline of the studied animal. The most promising applications of the LOUIS-3DM include 3D angiography, cancer research, and longitudinal studies of biological distribution of optoacoustic contrast agents (carbon nanotubes, metal plasmonic nanoparticles, etc.).
We developed the first prototype of dual-modality imager combining optoacoustic tomography (OAT) and laser
ultrasound tomography (UST) using computer models followed by experimental validation. The system designed
for preclinical biomedical research can concurrently yield images depicting both the absorbed optical energy
density and acoustic properties (speed of sound) of an object. In our design of the UST imager, we seek to
replace conventional electrical generation of ultrasound waves by laser-induced ultrasound (LU). While earlier
studies yielded encouraging results [Manohar, et al., Appl. Phys. Lett, 131911, 2007], they were limited to
two-dimensional (2D) geometries. In this work, we conduct computer-simulation studies to investigate different
designs for the 3D LU UST imager. The number and location of the laser ultrasound emitters, which are
constrained to reside on the cylindrical surface opposite to the arc of detectors, are optimized. In addition to
the system parameters, an iterative image reconstruction algorithm was optimized. We demonstrate that high
quality volumetric maps of the speed of sound can be reconstructed when only 32 emitters and 128 receiving
transducers are employed to record time-of-flight data at 360 tomographic view angles. The implications of the
proposed system for small animal and breast-cancer imaging are discussed.
In this report, we demonstrate the feasibility of using optoacoustic tomography for deducing biodistributions of
nanoparticles in animal models. The redistribution of single-walled carbon nanotubes (SWCNTs) was visualized in
living mice. Nanoparticle concentrations in harvested organs were measured spectroscopically using the intrinsic optical
absorption and fluorescence of SWCNTs. Observed increases in optoacoustic signal brightness in tissues were compared
with increases in optical absorptivity coefficients caused by SWCNT accumulation. The methodology presented in this
report paves the way for measuring concentrations of optically absorbing agents in small animals using optoacoustic
tomography.
We used a 3-D optoacoustic (OA) tomography system to create maps of optical absorbance of mice tissues contrasted with gold nanorods (GNRs). Nude mice were scanned before and after injection of GNRs at time periods varying from 1 to 192 h. Synthesized GNRs were purified from hexadecyltrimethylammonium bromide and coated with polyethylene glycol (PEG) to obtain GNR-PEG complexes suitable for in vivo applications. Intravenous administration of purified GNR-PEG complexes resulted in enhanced OA contrast of internal organs and blood vessels compared to the same mouse before injection of the contrast agent. Maximum enhancement of the OA images was observed 24 to 48 h postinjection, followed by a slow clearance trend for the remaining part of the studied period (eight days). We demonstrate that OA imaging with two laser wavelengths can be used for noninvasive, long-term studies of biological distribution of contrast agents.
Using the method of 3D optoacoustic tomography, we studied changes in tissues of the whole body of nude mice as the
changes manifested themselves from live to postmortem. The studies provided the necessary baseline for optoacoustic
imaging of necrotizing tissue, acute and chronic hypoxia, and reperfusion. They also establish a new optoacoustic model
of early postmortem conditions of the whole mouse body. Animals were scanned in a 37°C water bath using a three-dimensional
optoacoustic tomography system previously shown to provide high contrast maps of vasculature and organs
based on changes in the optical absorbance. The scans were performed right before, 5 minutes after, 2 hours and 1 day
after a lethal injection of KCl. The near-infrared laser wavelength of 765 nm was used to evaluate physiological features
of postmortem changes. Our data showed that optoacoustic imaging is well suited for visualization of both live and
postmortem tissues. The images revealed changes of optical properties in mouse organs and tissues. Specifically, we
observed improvements in contrast of the vascular network and organs after the death of the animal. We associated these
with reduced optical scattering, loss of motion artifacts, and blood coagulation.
Two-dimensional optoacoustic imaging with a hand-held probe operated in backward mode is being developed
for diagnostic imaging of breast cancer to evaluate the feasibility of a dual-modality optoacoustic plus ultrasonic
system that maps functional information of anatomical tissue structures with ultrasonic resolution. Tissue is
illuminated at 757nm and 1064nm for optical contrast between hypoxic blood of breast carcinomas and normally
oxygenated blood in benign masses. The system is optimized and calibrated in phantoms for a pilot clinical
study of patients with breast masses suspected for malignancy. Capability of the non-invasive system to improve
detection and diagnosis of breast tumors is discussed.
We report on the optoacoustic (OA) imaging of the whole mouse body using a biocompatible contrast agent - highly
purified, pegylated gold nanorods (GNR) - which has strong optical absorption in the near-infrared region and low level
of toxicity. In vitro toxicity studies showed no significant change in survival rates of the cultured normal epithelium
IEC-6 cells when incubated for 24 hours with up to 1 nM of GNR. In vivo toxicity studies in nude mice showed no
pathological changes in liver 1 month after the IV injection of GNR with intra-body concentration around 0.25-0.50 nM.
In order to study the enhancement of the OA contrast and accumulation of GNR in different tissues, we performed 3D
OA imaging of live nude mice with IV-injected GNR. The enhancement of the OA contrast in comparison with the
images of the untreated mice was visible starting 1 hour after the GNR injection. The OA contrast of kidneys, liver, and
spleen peaked at about 2-3 days after the administration of the GNR, and then was gradually reducing.
Coregistered optoacoustic (OA) and ultrasound (US) images obtained using a dual modality optoacoustic/ultrasonic
breast imaging system are used together for enhanced diagnostic capabilities in comparison to each individual
technology. Therefore, an operator-independent delineation of diagnostically relevant objects (in our case breast tumors)
with subsequent automatic analysis of image features is required. We developed the following procedure: 1) Image
filtering is implemented on a US image to minimize speckle noise and simultaneously preserve the sharpness of the
boundaries of the extended objects; 2) Boundaries of the objects of interest are automatically delineated starting with an
initial guess made by an operator; 3) Both US and OA images are analyzed using the detected boundaries (contrast,
boundary sharpness, homogeneity of the objects and background, boundary morphology parameters are calculated).
Calculated image characteristics can be used for statistically independent evaluation of structural information (US data)
and vascularization (OA data) of the studied breast tissues. Operator-independent delineation of the objects of interest
(e.g. tumors and blood vessels) is essential in clinical OA spectroscopy (using multiple laser wavelengths to quantify
concentrations of particular tissue chromophores, such as oxy- and deoxy- hemoglobin, water, and lipids). Another
potential application of the suggested image analysis algorithm could be in OA imaging system design, when system
performance should be evaluated in terms of quality of the images reconstructed from the well-defined objects of
interest. The discussed principles of image analysis are illustrated by using real clinical US and OA data.
We introduce a novel experimental design for non-invasive scanning optoacoustic microscopy that utilizes a parabolic
surface for ultrasound focusing. We demonstrate that off-axis parabolic mirrors made of sufficiently high acoustic
impedance materials work as ideal reflectors in a wide range of apertures and provide lossless conversion of a spherical
acoustic wavefront into a plane wave. We further test the performance of a custom optoacoustic imaging setup which
was developed and built based on these principles. The achieved resolution limit of 0.3 mm, with NA of 0.5 and the
transducer bandwidth of 5 MHz, matches the resolution limit defined by diffraction. Although further improvements of
current experimental setup are required to achieve resolution similar to leading microscopy systems, this proof-of-concept
work demonstrates the viability of the proposed design for optoacoustic microscopy applications.
We used a three-dimensional optical tomogaphy system that was previously developed to create high contrast maps of
optical absorbance of mice tissues. In this study, animals were scanned before and after injection of gold nanorods
(GNRs) at different time periods. As-synthesized GNRs were purified from hexadecyltrimethylammonium bromide
(CTAB) and coated with polyethylene glycol (PEG) to obtain GNR-PEG complexes suitable for in vivo applications.
Intravenous administration of the purified GNR-PEG complexes to mice resulted in an enhanced contrast of normal
tissues and blood vessels as compared to ordinary nude mice. In parallel with optoacoustic imaging we investigated the
accumulation of GNRs in liver using invasive analytical techniques. Maximum levels of GNRs in liver macrophages
were observed after 48-72 hours post-injection, followed by slow clearance trend after 8 days. Optoacoustic imaging
revealed redistribution of GNR in mouse organ and tissues: in the initial hours, accumulation of GNRs is seen
predominantly in the periphery of the mouse, while a gradual increase of GNR levels in liver, spleen and kidneys is seen
in 1 and 24 hours.
Optoacoustic (OA) tomography has demonstrated utility in identifying blood-rich malignancies in breast tissue. We describe the development and characterization of a laser OA imaging system for the prostate (LOIS-P). The system consists of a fiber-coupled Q-switched laser operating at 757 nm, a commercial 128-channel ultrasonic probe, a digital signal processor, and software that uses the filtered radial back-projection algorithm for image reconstruction. The system is used to reconstruct OA images of a blood-rich lesion induced in vivo in a canine prostate. OA images obtained in vivo are compared to images acquired using ultrasound, the current gold standard for guiding biopsy of the prostate. Although key structural features such as the urethra could be identified with both imaging techniques, a bloody lesion representing a highly vascularized tumor could only be clearly identified in OA images. The advantages and limitations of both forward and backward illumination modes are also evaluated by collecting OA images of phantoms simulating blood vessels within tissue. System resolution is estimated to be 0.2 mm in the radial direction of the acoustic array. The minimum detectable pressure signal is 1.83 Pa. Our results encourage further development toward a dual-modality OA/ultrasonic system for prostate imaging and image-guided biopsy.
A 3-D optoacoustic imaging system was used to visualize thermal lesions produced in vivo using high-intensity focused ultrasound (HIFU). A 7.5-MHz, surgical, focused transducer with a radius of curvature of 35 mm and an aperture diameter of 23 mm was used to generate HIFU. A pulsed laser, which could operate at 755 nm and 1064 nm, was used to illuminate excised tissue and mice using a bifurcated fiber bundle resulting in two wide beams of light. Tomographic images were obtained while the specimens were rotated within a sphere outlined by a concave arc-shaped array of 64 piezo-composite transducers. These images were then combined to reconstruct 3-D volume images (voxel resolution 0.5 mm), which were acquired before and after HIFU exposure. In vivo optoacoustic images acquired at 1064 nm provided visualization of HIFU lesions. The lesion was indicated by a negative optoacoustic contrast. The molecular nature of such contrast may possibly be associated with reduction of the optical absorption due to reduced concentration of blood, tissue dehydration, denaturation of proteins and porphyrins, and reduction of thermoacoustic efficiency in the thermally treated tissue. These preliminary results demonstrate the potential of optoacoustic imaging to assess and monitor the progress of HIFU therapy.
We present our findings from a real-time laser optoacoustic imaging system (LOIS). The system utilizes a Q-switched Nd:YAG laser; a standard 128-channel ultrasonic linear array probe; custom electronics and custom software to collect, process, and display optoacoustic (OA) images at 10 Hz. We propose that this system be used during preoperative mapping of forearm vessels for hemodialysis treatment. To demonstrate the real-time imaging capabilities of the system, we show OA images of forearm vessels in a volunteer and compare our results to ultrasound images of the same region. Our OA images show blood vessels in high contrast. Manipulations with the probe enable us to locate and track arteries and veins of a forearm in real time. We also demonstrate the ability to combine a series of OA image slices into a volume for spatial representation of the vascular network. Finally, we use frame-by-frame analysis of the recorded OA video to measure dynamic changes of the crossection of the ulnar artery.
In this work we modified light illumination of the laser optoacoustic (OA) imaging system to improve the 3D
visualization of human forearm vasculature. The computer modeling demonstrated that the new illumination design that
features laser beams converging on the surface of the skin in the imaging plane of the probe provides superior OA
images in comparison to the images generated by the illumination with parallel laser beams. We also developed the
procedure for vein/artery differentiation based on OA imaging with 690 nm and 1080 nm laser wavelengths. The
procedure includes statistical analysis of the intensities of OA images of the neighboring blood vessels. Analysis of the
OA images generated by computer simulation of a human forearm illuminated at 690 nm and 1080 nm resulted in
successful differentiation of veins and arteries. In vivo scanning of a human forearm provided high contrast 3D OA
image of a forearm skin and a superficial blood vessel. The blood vessel image contrast was further enhanced after it was
automatically traced using the developed software. The software also allowed evaluation of the effective blood vessel
diameter at each step of the scan. We propose that the developed 3D OA imaging system can be used during
preoperative mapping of forearm vessels that is essential for hemodialysis treatment.
A 3-D optoacoustic imaging system was used to visualize thermal lesions produced in excised tissue
specimens and in vivo mice using high intensity focused ultrasound (HIFU). A 7.5 MHz surgical,
focused transducer with a radius of curvature of 35 mm and an aperture diameter of 23 mm was used
to generate HIFU. A pulsed laser, which could operate at 755 nm and 1064 nm, was used to
illuminate excised tissue and mice using a bifurcated fiber bundle resulting in two wide beams of
light. Tomographic images were obtained while the specimens were rotated within a sphere outlined
by a concave arc-shaped array of 64 piezo-composite transducers. These images were then combined
to reconstruct 3-D volume images (voxel resolution 0.5 mm), which were acquired before and after
HIFU exposure. Optoacoustic images acquired at 1064 nm provided visualization of HIFU lesions.
The lesion in excised tissue was indicated by an increase in the optoacoustic signal; the in vivo lesion
was indicated by a decrease in the optoacoustic signal. The location and the extent of the lesions
were confirmed upon dissection. The discrepancy between the ex vivo and the in vivo results might
be attributed to the different effective thermal deposition in the two cases. These preliminary results
demonstrate the potential of optoacoustic imaging to assess and monitor the progress of HIFU
therapy.
We present results from a clinical case study on imaging breast cancer using a real-time interleaved two laser
optoacoustic imaging system co-registered with ultrasound. The present version of Laser Optoacoustic Ultrasonic
Imaging System (LOUIS) utilizes a commercial linear ultrasonic transducer array, which has been modified to include
two parallel rectangular optical bundles, to operate in both ultrasonic (US) and optoacoustic (OA) modes. In OA mode,
the images from two optical wavelengths (755 nm and 1064 nm) that provide opposite contrasts for optical absorption of
oxygenated vs deoxygenated blood can be displayed simultaneously at a maximum rate of 20 Hz. The real-time aspect of
the system permits probe manipulations that can assist in the detection of the lesion. The results show the ability of
LOUIS to co-register regions of high absorption seen in OA images with US images collected at the same location with
the dual modality probe. The dual wavelength results demonstrate that LOUIS can potentially provide breast cancer
diagnostics based on different intensities of OA images of the lesion obtained at 755 nm and 1064 nm. We also present
new data processing based on deconvolution of the LOUIS impulse response that helps recover original optoacoustic
pressure profiles. Finally, we demonstrate the image analysis tool that provides automatic detection of the tumor
boundary and quantitative metrics of the optoacoustic image quality. Using a blood vessel phantom submerged in a
tissue-like milky background solution we show that the image contrast is minimally affected by the phantom distance
from the LOUIS probe until about 60-65 mm. We suggest using the image contrast for quantitative assessment of an OA
image of a breast lesion, as a part of the breast cancer diagnostics procedure.
In this report we demonstrate improved three-dimensional optoacoustic tomography in test samples. High quality
tomographic data and images were obtained from phantom of mice being 2.5 cm in diameter. Capillaries filled with
cupric sulfate, ferrous sulfate and nickel sulfate solutions, and immersed in a scattering medium were used for these
tests. The brightness of reconstructed phantom images was found to match accurately the absorption profiles of test
solutions. Hence, optoacoustic imaging can be applied in preclinical research to perform in vivo absorptivity
measurements to deduce functional information on blood oxygen levels or concentration of contrast agents.
Optoacoustic Tomography (OAT) is a hybrid imaging modality that combines the advantages of both optical
imaging and ultrasound imaging techniques. Most existing reconstruction algorithms for OAT assume pointlike
transducers, which may result in conspicous image blurring and distortions in certain applications. In this
work, a new imaging model that incorporates the transducer response is employed for image reconstruction.
Computer-simulation studies demonstrate that the new reconstruction method can effectively compensate for
image resolution degradation associated with the transducer response.
We develop a system for three-dimensional whole-body optoacoustic tomography of small animals for applications in preclinical research. The tomographic images are obtained while the objects of study (phantoms or mice) are rotated within a sphere outlined by a concave arc-shaped array of 64 piezocomposite transducers. Two pulsed lasers operating in the near-IR spectral range (755 and 1064 nm) with an average pulsed energy of about 100 mJ, a repetition rate of 10 Hz, and a pulse duration of 15 to 75 ns are used as optical illumination sources. During the scan, the mouse is illuminated orthogonally to the array with two wide beams of light from a bifurcated fiber bundle. The system is capable of generating images of individual organs and blood vessels through the entire body of a mouse with spatial resolution of ~0.5 mm.
KEYWORDS: Transducers, Acoustics, Sensors, Calibration, Wavefronts, Ultrasonography, Signal attenuation, Pulsed laser operation, Image restoration, Signal to noise ratio
Comprehensive characterization of optoacoustic transducers is achieved through the analysis of their frequency
response using a procedure of measuring angular dependence of the transducer sensitivity to the ultrawide-band
delta pulse. The testing was performed under standard repeatable operating conditions. Back-illumination of a
blackened, acoustically matched, planar surface with a short laser pulse creates an acoustic impulse which was used
as an ultrawide-band ultrasonic source. The bandwidth of such a source extends well over 10 MHz (6dB point at 16
MHz for illumination with a 16 ns pulse) and the low frequency
roll-off is around 300 kHz. Analysis of the angular
dependence of the frequency response yields invaluable directivity information about the detector under study,
which in turn permits accurate forward and inverse problem models.
We developed a 3D whole-body optoacoustic tomography system for applications in preclinical research on mice. The
system is capable of generating images with resolution better than 0.6 mm. Two pulsed lasers, an Alexandrite laser
operating at 755 nm and a Nd:YAG laser operating at 532 nm and 1064nm were used for light delivery. The
tomographic images were obtained while the objects of study (phantoms or mice) were rotated within a sphere outlined
by a concave arc-shaped array of 64 piezo-composite transducers. During the scan, the mouse was illuminated
orthogonally to the array with two wide beams of light from a bifurcated fiber bundle. Illumination at 532 nm showed
superficial vasculature, but limited penetration depth at this wavelength prevented the detection of deeper structures.
Illumination at 755 and 1064 nm showed organs and blood vessels, respectively. Filtering of the optoacoustic signals
using high frequency enhancing wavelets further emphasized the smaller blood vessels.
We describe two laser optoacoustic imaging systems for breast cancer detection based on arrays of acoustic detectors
operated manually in a way similar to standard ultrasonic breast imaging. The systems have the advantages of standard
light illumination (regardless of the interrogated part of the breast), the ability to visualize any part of the breast, and
convenience in operation. The first system could work in both ultrasonic and optoacoustic mode, and was developed
based on a linear ultrasonic breast imaging probe with two parallel rectangular optical bundles. We used it in a pilot
clinical study to provide for the first time demonstration that the boundaries of the tumors visualized on the optoacoustic
and ultrasonic images matched. Such correlation of coregistered images proves that the objects on both images
represented indeed the same tumor. In the optoacoustic mode we were also able to visualize blood vessels located in the
neighborhood of the tumor. The second system was proposed as a circular array of acoustic transducers with an axisymmetric
laser beam in the center. It was capable of 3D optoacoustic imaging with minimized optoacoustic artifacts
caused by the distribution of the absorbed optical energy within the breast tissue. The distribution of optical energy
absorbed in the bulk tissue of the breast was removed from the image by implementing the principal component analysis
on the measured signals. The computer models for optoacoustic imaging using these two handheld probes were
developed. The models included three steps: (1) Monte Carlo simulations of the light distribution within the breast
tissue, (2) generation of optoacoustic signals by convolving
N-shaped pressure signals from spherical voxels with the
shape of individual transducers, and (3) back-projecting processed optoacoustic signals onto spherical surfaces for image
reconstruction. Using the developed models we demonstrated the importance of the included spatial impulse response of
the optoacoustic imaging system.
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