A review of a strategic roadmapping exercise to advance clinical translation of photoacoustic imaging: From current barriers to future adoption.

Photoacoustic imaging (PAI), also referred to as optoacoustic imaging, has shown promise in early-stage clinical trials in a range of applications from inflammatory diseases to cancer. While the first PAI systems have recently received regulatory approvals, successful adoption of PAI technology into healthcare systems for clinical decision making must still overcome a range of barriers, from education and training to data acquisition and interpretation. The International Photoacoustic Standardisation Consortium (IPASC) undertook an community exercise in 2022 to identify and understand these barriers, then develop a roadmap of strategic plans to address them. Here, we outline the nature and scope of the barriers that were identified, along with short-, medium- and long-term community efforts required to overcome them, both within and beyond the IPASC group.

Stochastic three-dimensional numerical phantoms to enable computational studies in quantitative optoacoustic computed tomography of breast cancer.

Significance: When developing a new quantitative optoacoustic computed tomography (OAT) system for diagnostic imaging of breast cancer, objective assessments of various system designs through human trials are infeasible due to cost and ethical concerns. In prototype stages, however, different system designs can be cost-efficiently assessed via virtual imaging trials (VITs) employing ensembles of digital breast phantoms, i.e., numerical breast phantoms (NBPs), that convey clinically relevant variability in anatomy and optoacoustic tissue properties. Aim: The aim is to develop a framework for generating ensembles of realistic three-dimensional (3D) anatomical, functional, optical, and acoustic NBPs and numerical lesion phantoms (NLPs) for use in VITs of OAT applications in the diagnostic imaging of breast cancer. Approach: The generation of the anatomical NBPs was accomplished by extending existing NBPs developed by the U.S. Food and Drug Administration. As these were designed for use in mammography applications, substantial modifications were made to improve blood vasculature modeling for use in OAT. The NLPs were modeled to include viable tumor cells only or a combination of viable tumor cells, necrotic core, and peripheral angiogenesis region. Realistic optoacoustic tissue properties were stochastically assigned in the NBPs and NLPs. Results: To advance optoacoustic and optical imaging research, 84 datasets have been released; these consist of anatomical, functional, optical, and acoustic NBPs and the corresponding simulated multi-wavelength optical fluence, initial pressure, and OAT measurements. The generated NBPs were compared with clinical data with respect to the volume of breast blood vessels and spatially averaged effective optical attenuation. The usefulness of the proposed framework was demonstrated through a case study to investigate the impact of acoustic heterogeneity on OAT images of the breast. Conclusions: The proposed framework will enhance the authenticity of virtual OAT studies and can be widely employed for the investigation and development of advanced image reconstruction and machine learning-based methods, as well as the objective evaluation and optimization of the OAT system designs.

Normalization of optical fluence distribution for three-dimensional functional optoacoustic tomography of the breast.

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.

Opto-acoustic imaging of relative blood oxygen saturation and total hemoglobin for breast cancer diagnosis.

Opto-acoustic imaging involves using light to produce sound waves for visualizing blood in biological tissue. By using multiple optical wavelengths, diagnostic images of blood oxygen saturation and total hemoglobin are generated using endogenous optical contrast, without injection of any external contrast agent and without using any ionizing radiation. The technology has been used in recent clinical studies for diagnosis of breast cancer to help distinguish benign from malignant lesions, potentially reducing the need for biopsy through improved diagnostic imaging accuracy. To enable this application, techniques for mapping oxygen saturation differences within tissue are necessary. Using biologically relevant opto-acoustic phantoms, we analyze the ability of an opto-acoustic imaging system to display colorized parametric maps that are generated using a statistical mapping approach. To mimic breast tissue, a material with closely matching properties for optical absorption, optical scattering, acoustic attenuation, and speed of sound is used. The phantoms include two vessels filled with whole blood at oxygen saturation levels determined using a sensor-based approach. A flow system with gas-mixer and membrane oxygenator adjusts the oxygen saturation of each vessel independently. Datasets are collected with an investigational Imagio® breast imaging system. We examine the ability to distinguish vessels as the oxygen saturation level and imaging depth are varied. At depth of 15 mm and hematocrit of 42%, a sufficient level of contrast to distinguish between two 1.6-mm diameter vessels was measured for an oxygen saturation difference of ∼4.6 % . In addition, an oxygenated vessel was visible at a depth of 48 mm using an optical wavelength of 1064 nm, and a deoxygenated vessel was visible to a depth of 42 mm with 757 nm. The results provide insight toward using color mapped opto-acoustic images for diagnosing breast cancer.

Special Section Guest Editorial: Celebrating the Exponential Growth of Optoacoustic/Photoacoustic Imaging.

Guest editors introduce contributors to the Special Section Celebrating the Exponential Growth of Optoacoustic/Photoacoustic Imaging.

Temperature-dependent optoacoustic response and transient through zero Grüneisen parameter in optically contrasted media.

Non-invasive optoacoustic mapping of temperature in tissues with low blood content can be enabled by administering external contrast agents. Some important clinical applications of such approach include temperature mapping during thermal therapies in a prostate or a mammary gland. However, the technique would require a calibration that establishes functional relationship between the measured normalized optoacoustic response and local tissue temperature. In this work, we investigate how a key calibration parameter - the temperature of zero optoacoustic response (T0 ) - behaves in different environments simulating biological tissues augmented with either dissolved or particulate (nanoparticles) contrast agents. The observed behavior of T0 in ionic and molecular solutions suggests that in-vivo temperature mapping is feasible for contrast agents of this type, but requires knowledge of local concentrations. Oppositely, particulate contrast agents (plasmonic or carbon nanoparticles) demonstrated concentration-independent thermal behavior of optoacoustic response with T0 defined by the thermoelastic properties of the local environment.

Impact of nonstationary optical illumination on image reconstruction in optoacoustic tomography.

Optoacoustic tomography (OAT), also known as photoacoustic tomography, is a rapidly emerging hybrid imaging technique that possesses great potential for a wide range of biomedical imaging applications. In OAT, a laser is employed to illuminate the tissue of interest and acoustic signals are produced via the photoacoustic effect. From these data, an estimate of the distribution of the absorbed optical energy density within the tissue is reconstructed, referred to as the object function. This quantity is defined, in part, by the distribution of light fluence within the tissue that is established by the laser source. When performing three-dimensional imaging of large objects, such as a female human breast, it can be difficult to achieve a relatively uniform coverage of light fluence within the volume of interest when the position of the laser source is fixed. To circumvent this, researchers have proposed illumination schemes in which the relative position of the laser source and ultrasound probe is fixed, and both are rotated together to acquire a tomographic dataset. A problem with this rotating-illumination scheme is that the tomographic data are inconsistent; namely, the acoustic data recorded at each tomographic view angle (i.e., probe position) are produced by a distinct object function. In this work, the impact of this data inconsistency on image reconstruction accuracy is investigated systematically. This is accomplished by use of computer-simulation studies and application of mathematical results from the theory of microlocal analysis. These studies specify the set of image discontinuities that can be stably reconstructed with a nonstationary optical illumination setup. The study also includes a comparison of the ability of iterative and analytic image reconstruction methods to mitigate artifacts attributable to the data inconsistency.

All-optical optoacoustic microscopy based on probe beam deflection technique.

Optoacoustic (OA) microscopy using an all-optical system based on the probe beam deflection technique (PBDT) for detection of laser-induced acoustic signals was investigated as an alternative to conventional piezoelectric transducers. PBDT provides a number of advantages for OA microscopy including (i) efficient coupling of laser excitation energy to the samples being imaged through the probing laser beam, (ii) undistorted coupling of acoustic waves to the detector without the need for separation of the optical and acoustic paths, (iii) high sensitivity and (iv) ultrawide bandwidth. Because of the unimpeded optical path in PBDT, diffraction-limited lateral resolution can be readily achieved. The sensitivity of the current PBDT sensor of 22 µV/Pa and its noise equivalent pressure (NEP) of 11.4 Pa are comparable with these parameters of the optical micro-ring resonator and commercial piezoelectric ultrasonic transducers. Benefits of the present prototype OA microscope were demonstrated by successfully resolving micron-size details in histological sections of cardiac muscle.

Optical clearing of skin enhanced with hyaluronic acid for increased contrast of optoacoustic imaging.

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.

"Contrast agents for optoacoustic imaging: design and biomedical applications".

Nanoparticles and molecular chromophores with strong optical absorption in the near-infrared spectral range can be used as contrast agents for optoacoustic (photoacoustic) imaging, thereby significantly enhancing sensitivity and enabling new applications of this novel and rapidly growing biomedical imaging technology.

Melanin nanoparticles as a novel contrast agent for optoacoustic tomography.

We describe the synthesis and characterization of melanin-like nanoparticles (MNP) as novel contrast agents for optoacoustic tomography. Good dispersion stability of high concentration MNPs in different biological media was achieved with thiol-terminated methoxy-poly(ethylene glycol), which can be used for further functional conjugation. MNP-PEG were found biocompatible with human MCF-7 and 3T3 cells. Cell toxicity of MNPs was found lower than that of gold nanorods for concentrations that provide equal optical absorbance. Optoacoustic tomography images were obtained with Laser Optoacoustic Imaging System (LOIS-3D) from tubes filled with contrast agents and live mice. Imaging of tubes permitted verification of the system resolution <300 µm and sensitivity Δµa=0.03/cm under safe laser fluence of 20 mJ/cm(2). Water suspensions of MNP demonstrated optoacoustic efficiency that is about equal to that of gold nanorods under conditions of equal optical absorption. We conclude that MNPs have the potential for biomedical imaging applications as optoacoustic contrast agents.

Red blood cell as a universal optoacoustic sensor for non-invasive temperature monitoring.

Optoacoustic (photoacoustic) temperature imaging could provide improved spatial resolution and temperature sensitivity as compared to other techniques of non-invasive thermometry used during thermal therapies for safe and efficient treatment of lesions. However, accuracy of the reported optoacoustic methods is compromised by biological variability and heterogeneous composition of tissues. We report our findings on the universal character of the normalized temperature dependent optoacoustic response (ThOR) in blood, which is invariant with respect to hematocrit at the isosbestic point of hemoglobin. The phenomenon is caused by the unique homeostatic compartmentalization of blood hemoglobin exclusively inside erythrocytes. On the contrary, the normalized ThOR in aqueous solutions of hemoglobin showed linear variation with respect to its concentration and was identical to that of blood when extrapolated to the hemoglobin concentration inside erythrocytes. To substantiate the conclusions, we analyzed optoacoustic images acquired from the samples of whole and diluted blood as well as hemoglobin solutions during gradual cooling from +37 to -15 °C. Our experimental methodology allowed direct observation and accurate measurement of the temperature of zero optoacoustic response, manifested as the sample's image faded into background and then reappeared in the reversed (negative) contrast. These findings provide a framework necessary for accurate correlation of measured normalized optoacoustic image intensity and local temperature in vascularized tissues independent of tissue composition.

Enabling in vivo measurements of nanoparticle concentrations with three-dimensional optoacoustic tomography.

In this report, we demonstrate the feasibility of using optoacoustic tomography (OAT) to evaluate 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 absorption coefficients caused by SWCNT accumulation. The methodology presented in this report can further be extended to calibrate the sensitivity of an optoacoustic imaging system for a range of changes in optical absorption coefficient values at specific locations or organs in a mouse body to enable noninvasive measurements of nanoparticle concentrations in vivo. Additionally, qualitative information provided by OAT and quantitative information obtained ex vivo may provide valuable feedback for advancing methods of quantitative analysis with OAT.

Accelerating image reconstruction in three-dimensional optoacoustic tomography on graphics processing units.

PURPOSE: Optoacoustic tomography (OAT) is inherently a three-dimensional (3D) inverse problem. However, most studies of OAT image reconstruction still employ two-dimensional imaging models. One important reason is because 3D image reconstruction is computationally burdensome. The aim of this work is to accelerate existing image reconstruction algorithms for 3D OAT by use of parallel programming techniques. METHODS: Parallelization strategies are proposed to accelerate a filtered backprojection (FBP) algorithm and two different pairs of projection/backprojection operations that correspond to two different numerical imaging models. The algorithms are designed to fully exploit the parallel computing power of graphics processing units (GPUs). In order to evaluate the parallelization strategies for the projection/backprojection pairs, an iterative image reconstruction algorithm is implemented. Computer simulation and experimental studies are conducted to investigate the computational efficiency and numerical accuracy of the developed algorithms. RESULTS: The GPU implementations improve the computational efficiency by factors of 1000, 125, and 250 for the FBP algorithm and the two pairs of projection/backprojection operators, respectively. Accurate images are reconstructed by use of the FBP and iterative image reconstruction algorithms from both computer-simulated and experimental data. CONCLUSIONS: Parallelization strategies for 3D OAT image reconstruction are proposed for the first time. These GPU-based implementations significantly reduce the computational time for 3D image reconstruction, complementing our earlier work on 3D OAT iterative image reconstruction.

Three-dimensional optoacoustic imaging as a new noninvasive technique to study long-term biodistribution of optical contrast agents in small animal models.

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.

Highly purified biocompatible gold nanorods for contrasted optoacoustic imaging of small animal models.

We developed a methodology for high yield synthesis of gold nanorods (GNR) with narrow band optical absorption centered at 760 nm. GNR were purified from hexadecyltrimethylammonium bromide (CTAB) and coated with polyethylene glycol (PEG). The molar ratio between GNR and PEG (1÷50000) was optimized to make the conjugate a biocompatible PEG-GNR contrast agent for optoacoustic (OA) imaging. In vitro toxicity studies showed no significant change in survival rates of cultured normal (IEC-6, MDCK) and cancer (SKBR3 and HEPG2) cells after they were incubated with 0.125 to 1.25 nM PEG-GNR solutions. In vivo toxicity studies in nude mice showed no pathological changes in liver after the IV injection of GNR. Significant enhancements of OA contrast in comparison to images of untreated mice were observed 1 hour after the GNR injection in a dose of 20 mg gold per kg of body mass.

Investigation of iterative image reconstruction in three-dimensional optoacoustic tomography.

Iterative image reconstruction algorithms for optoacoustic tomography (OAT), also known as photoacoustic tomography, have the ability to improve image quality over analytic algorithms due to their ability to incorporate accurate models of the imaging physics, instrument response and measurement noise. However, to date, there have been few reported attempts to employ advanced iterative image reconstruction algorithms for improving image quality in three-dimensional (3D) OAT. In this work, we implement and investigate two iterative image reconstruction methods for use with a 3D OAT small animal imager: namely a penalized least-squares (PLS) method employing a quadratic smoothness penalty and a PLS method employing a total variation norm penalty. The reconstruction algorithms employ accurate models of the ultrasonic transducer impulse responses. Experimental data sets are employed to compare the performances of the iterative reconstruction algorithms to that of a 3D filtered backprojection (FBP) algorithm. By the use of quantitative measures of image quality, we demonstrate that the iterative reconstruction algorithms can mitigate image artifacts and preserve spatial resolution more effectively than FBP algorithms. These features suggest that the use of advanced image reconstruction algorithms can improve the effectiveness of 3D OAT while reducing the amount of data required for biomedical applications.

Laser nanothermolysis of human leukemia cells using functionalized plasmonic nanoparticles.

In the present work, we present the use of gold nanorods as plasmonic nanoparticles for selective photothermal therapy of human acute (HL-60) and chronicle (K-562) leukemia cells using a near-infrared laser. We improved a published methodology of gold nanorods conjugation to generate high yields of narrow band gold nanorods with an optical absorption centered at 760 nm. The manufactured nanorods were pegylated and conjugated with monoclonal antibody to become non-toxic as biocompatible nanothermolysis agent. Gold nanorods are synthesized and conjugated to CD33 monoclonal antibody. After pegylation, or conjugation with CD33 antibody, gold nanorods were non-toxic to acute and chronic leukemia cells. Our modified gold nanorods CD33 conjugates shown high level of accumulation for both leukemia cell lines, and successful used for nanothermolysis of human leukemia cells in vitro. Each sample was illuminated with 1 or 3 laser shots as for low and for high laser fluence. The radiation was provided by a Quanta Systems q-switched titanium sapphire laser, and the system was designed for maximum sample coverage using non-focused illumination. HL-60 and K-562 cells were treated for 45 min with gold nanorods CD33 conjugated, or with pegylated gold nanorods. The effect of pulsed-laser nanothermolysis for acute and chronic leukemia cells were investigated with cell counting for number of living cells, percentage of cell death and functional parameters such as damage of cell membrane and metabolic activity. Gold nanorods CD33 conjugates significantly increase cell damage for low fluence laser and completely destroyed cancer cells after 3 pulses for low fluence (acute leukemia) and for high fluence laser as for HL-60 (acute) and for K-562 (chronicle) leukemia cells.

An imaging model incorporating ultrasonic transducer properties for three-dimensional optoacoustic tomography.

Optoacoustic tomography (OAT) is a hybrid imaging modality that combines the advantages of optical and ultrasound imaging. Most existing reconstruction algorithms for OAT assume that the ultrasound transducers employed to record the measurement data are point-like. When transducers with large detecting areas and/or compact measurement geometries are utilized, this assumption can result in conspicuous image blurring and distortions in the reconstructed images. In this work, a new OAT imaging model that incorporates the spatial and temporal responses of an ultrasound transducer is introduced. A discrete form of the imaging model is implemented and its numerical properties are investigated. We demonstrate that use of the imaging model in an iterative reconstruction method can improve the spatial resolution of the optoacoustic images as compared to those reconstructed assuming point-like ultrasound transducers.