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Optoacoustic Imaging is based on the thermal expansion of tissue caused by a temperature rise due to absorption of short laser pulses. At constant laser fluence, optoacoustic image contrast is proportional to differences in optical absorption and the thermoacoustic efficiency, expressed by the Grneisen parameter, Γ. Γ is proportional to the thermal expansion coefficient, the sound velocity squared and the inverse heat capacity at constant pressure. In thermal therapies, these parameters may be modified in the treated area. In this work experiments were performed to examine the influence of these parameters on image contrast. A Laser Optoacoustic Imaging System (LOIS, Fairway Medical Technologies, Houston, Texas) was used to image tissue phantoms comprised of cylindrical Polyvinyl Chloride Plastisol (PVCP) optical absorbing targets imbedded in either gelatin or PVCP as the background medium. Varying concentrations of Black Plastic Color (BPC) and titanium dioxide (TiO2)were added to targets and background to yield desired tissue relevant optical absorption and effective scattering coefficients, respectively. In thermal therapy experiments, ex-vivo bovine liver was heated with laser fibres (805nm laser at 5 W for 600s) to create regions of tissue coagulation. Lesions formed in the liver tissue were visible using the LOIS system with reasonable correspondence to the actual region of tissue coagulation. In the phantom experiments, contrast could be seen with low optical absorbing targets (μaof 0.50cm−1down to0.13cm−1)embedded in a gelatin background (μa=0.13cm−1andμs′=4.2cm−1). Therefore, the data suggest that small objects (

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 22μV/Pa and its noise equivalent pressure (NEP) of 11.4Pa 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.

One area of particular relevance to optoacoustic imaging is the enhancement of malignant tumor contrast relative to background normal tissue. Although optoacoustic imaging has been shown to be superior to the commonly employed clinical imaging modalities in terms of contrast, optoacoustic contrast agents offer the possibility of both visualizing and treating currently invisible pathological entities. It is likely that operation of a variety of imaging and monitoring devices utilizing optoacoustic principles may be enhanced with application of contrast agents based on metal nanoparticles (NPs) possessing plasmon resonance absorption. Gold and silver NPs will be homogeneously heated with the nanosecond pulses of Q-switched lasers commonly used in optoacoustic imaging and sensing. Solid gold and silver nanorods (NRs) not only possess the strongest optical absorption, the peak of this absorption can be conveniently tuned by making them longer, while keeping their diameter constant.

Noninvasive determination of optical energy distribution in live biological tissues is an important aspect of any laser treatment or diagnostic procedure. It is particularly important for optical dosimetry in tissue, tissue diagnostics based on optical properties, quantitative molecular and functional imaging in the human body based on optical contrast, laser therapeutic procedures (such as photodynamic therapy, photothermal therapy, and laser treatment of various skin lesions and hair removal), and laser microsurgery. Monitoring of changes in tissue optical properties can also be utilized for monitoring of laser–tissue interactions, noninvasive optical biopsy, the effects of laser ablation or denaturation of tissue, and ablation by high-intensity focused ultrasound (HIFU). The precision of these procedures would improve significantly if laser irradiation conditions could be adjusted in accordance with initial optical properties and with changes in optical properties during the course of the treatment. Therefore, there is a definite need for noninvasive measurement and visualization of the light distribution profile in the laser-irradiated tissue in vivo. Such feedback information helps predict the optimal laser wavelength, pulse duration, laser intensity, and light dose to be delivered so as to obtain diagnostic information or to cause a sufficient effect during treatment with minimal damage to adjacent tissue layers.

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 (

An optoacoustic (OA) sensor was designed, fabricated, and used to detect spherical gold nanoparticles (NPs) in diluted suspensions. The sensor, operating in the backward mode, was designed to measure signals from microscopic volumes of nanoparticulate suspensions in water. Thermal nonlinearity was observed in the course of OA signal generation. The irradiation of a microvolume of gold nanoparticles at the wavelength matching the peak of their plasmon resonance absorption gives rise to a multitude of thermomechanical processes, including heating of NPs below the critical temperature of water (374 K). The thermal diffusion from nanoparticles to water takes place; however, formation of vapor nanobubbles is avoided. As a result, a specific acoustic signal is produced exhibiting nonlinear behavior with respect to the incident laser pulse energy. The optoacoustic profile of the laser-induced signal generated in a thin layer of highly diluted suspensions of gold nanospheres was examined, thereby providing a basis for a method for detection of metal nanoparticles with high sensitivity.

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 data set. 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 non-stationary optical illumination set-up. The study also includes a comparison of the ability of iterative and analytic image reconstruction methods to mitigate artifacts attributable to the data inconsistency.

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 and other type of biomedical research. The system (LOUIS-3DM) combines a multiwavelength optoacoustic tomography (OAT) and laser-induced ultrasound tomography (LUT) to obtain coregistered maps of tissue optical absorption and speed of sound, 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 distributions of optoacoustic contrast agents.

Optoacoustic tomography (OAT) is a promising imaging modality for human breast cancer imaging, with higher resolution and deeper penetration compared to other optical imaging modalities such as diffuse optical tomography or optical coherence tomography. It yields a resolution of 1 mm at depth up to 2 cm. But there is an inherent conflict between the limitations imposed on laser power and the need to adequately penetrate a substantial portion of the breast. To achieve sufficient penetration at every view angle, instead of illuminating the whole breast all at once, sometimes illumination is focused onto a small region of the breast and rotated along with the transducer array to cover the entire object. This paper evaluates the effect of this rotating partial illumination design on OAT image reconstruction. The optical process is simulated by conducting Monte Carlo simulations on a numerical phantom mimicking a real breast, with various specially designed illumination schemes. The acoustic process is simulated by incorporating the transducer's spatial impulse response. Iterative reconstruction is applied to estimate the OAT image. We conclude that rotating partial illumination introduces inconsistency into the system equation, and the degree of inconsistency determines the reconstruction quality.

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.).

Optoacoustic (OA) sensor was designed, fabricated and used to detect spherical gold nanoparticles (NPs) in suspensions at the level of up to a single particle. The sensor operating in the backward mode was designed to measure signals from microscopic volumes of nanoparticulate suspensions in water. Thermal nonlinearity was observed in the course of OA signal generation. The irradiation of the microvolume of gold nanoparticles gives rise to a multitude of thermomechanical processes, including heating of NPs above the vaporization temperature, thermal diffusion from nanoparticles to water and formation of vapor nanobubbles. As a result, a specific acoustic signal is produced exhibiting nonlinear behaviour with respect to the incident laser pulse energy. The optoacoustic profile of the laser-induced signal generated in a thin layer of highly diluted suspensions of gold nanospheres was examined thereby providing a basis for a method for detection of metal nanoparticles with high sensitivity.

We describe novel ex vivo method for elimination of tumor cells from cell suspension, Laser Activated Nanothermolysis and propose this method for purging of bone marrow and blood transplants. K562 and human lympholeukemia cells were eliminated in experiments by laser-induced micro-bubbles that emerge inside individual target cells around selectively formed clusters of light-absorbing gold nanoparticles. Pretreatment of tumor cells with specific monoclonal antibodies and Ig-conjugated 30-nm gold particles allowed the formation of clusters of 10-20 on the surface of cell membrane. Electron microscopy found the nanoparticulate clusters inside the cells. Total (100%) elimination of K562 cells targeted with specific antibodies was achieved with single laser pulses with optical fluence of 5J/cm(2) at the wavelength of 532 nm without damage to the same cells targeted without specific antibodies. Total elimination of human lymphoblasts from suspension of normal stem cells was achieved by a single laser pulse with the optical fluence of 1.7J/cm(2), while the damage level of normal cells was 16%.

Background and Objective Previously reported studies on laser nano-thermolysis of cancerous cells demonstrated insufficient efficacy and specificity of malignant cell damage. Safety, that is, absence of damage to normal cells in the course of the laser thermolysis was also low due to less than optimal protocol of cancer cell targeting with nanoparticles (NP). The objective of this study was two-fold: to optimize NP targeting to real tumor (human) cells and to better understand physical mechanisms of cell damage for improved control of the laser ablation. Study Design/Materials and Methods We have suggested (1) two-stage targeting method to form clusters of light-absorbing gold NPs selectively in target cells, and (2) the cell damage mechanism through laser-induced generation of vapor bubbles around NP clusters. Experimental investigation of laser nano-thermolysis of leukemia cells was performed using 30 nm spherical gold nanoparticles as a light absorbing agent, and photothermal and fluorescent microscopies as well as flow cytometry as methods to monitor microbubble formation and resulting damage of leukemia cells in human bone marrow specimens. Results NP clusters were formed and visualized using fluorescence microscopy at cell membranes and in cytoplasm of B-lymphoblasts. Laser irradiation of cells (532 nm, 10 nanoseconds, 0.6 J/cm2) induced microbubbles selectively in leukemia cells with large clusters, but not in cells with single NPs or small clusters. Quantitative analysis demonstrated that only 0.1%–1.5% of tumor cells and 77%–84% of normal bone marrow cells survived laser pulse. Conclusions Two-stage cell targeting method permits formation of NP clusters selectively in diagnosis-specific tumor cells. The clusters serve as effective sources of photothermally-induced microbubbles, which kill individual target cells after a single laser pulse. The laser fluence threshold for generation of microbubbles is inversely proportional to the volume of NP clusters. Lasers Surg. Med. 38:631–642, 2006. © 2006 Wiley-Liss, Inc.

A laser optoacoustic imaging system (LOIS) uses time-resolved detection of laser-induced pressure profiles in tissue in order to reconstruct images of the tissue based on distribution of acoustic sources. Laser illumination with short pulses generates distribution of acoustic sources that accurately replicates the distribution of absorbed optical energy. The complex spatial profile of heterogeneous distribution of acoustic sources can be represented in the frequency domain by a wide spectrum of ultrasound ranging from tens of kilohertz to tens of megahertz. Therefore, LOIS requires a unique acoustic detector operating simultaneously within a wide range of ultrasonic frequencies. Physical principles of an array of ultrawide-band ultrasonic transducers used in LOIS designed for imaging tumors in the depth of tissue are described. The performance characteristics of the transducer array were modeled and compared with experiments performed in gel phantoms resembling optical and acoustic properties of human tissue with small tumors. The amplitude and the spectrum of laser-induced ultrasound pulses were measured in order to determine the transducer sensitivity and the level of thermal noises within the entire ultrasonic band of detection. Spatial resolution of optoacoustic images obtained with an array of piezoelectric transducers and its transient directivity pattern within the field of view are described. The detector design considerations essential for obtaining high-quality optoacoustic images are presented.

The spatial distribution of laser radiation intensity in turbid condensed media is studied analytically, numerically, and experimentally by the optoacoustic method. Based on optoacoustic measurements and Monte Carlo numerical simulations, the relation is obtained between the optical characteristics of a scattering medium and the position of the maximum of the spatial distribution of radiation intensity in the medium. It is shown that, when the anisotropy factor exceeds 0.8, this dependence has a universal type in the ranges of absorption and scattering coefficients typical for biological tissues. The method is proposed for measuring the extinction and absorption coefficients in light scattering media from the temporal shape of an optoacoustic pulse detected in relative units. An approximate method for solving a radiation transfer equation is verified, and the regions of application of the P3 and P5 approximation are established.

Optoacoustic microscopy (OAM) is an emerging technology combining the beneficial features of optical contrast and ultrasound resolution, to form a hybrid imaging technique capable of multi-scale, high-contrast and high-resolution imaging through optically scattering biological tissues. In the past 15 years, two system modifications have been developed for optoacoustic / photoacoustic microscopy: acoustic-resolution AR-OAM and optical-resolution OR-OAM. Typically, acoustic resolution systems can image deeper tissues structures, however, with resolution at least an order of magnitude worse than the systems of optical-resolution. It would be attractive for variety of biomedical applications to attain high (submicron) resolution at a depth exceeding the present limit of the optical resolution optoacoustic microscopy. Here we introduce a novel, all-optical method for OAM, in which not only thermal energy deposition, but also optoacoustic signal detection is achieved optically. In our design the probe laser beam was used as an ultrawide-band ultrasonic transducer. In this method the acoustic pressure wave amplitude is proportional to the angle of deflection of the probing CW laser beam incident on a balanced dual photodiode. Such laser beam deflection (LBD) method overcomes the limitations of conventional piezoelectric ultrasound transducers and optical interferometers. LBD method allows one to use high numerical aperture objectives for better focusing, avoid distortions associated with the system elements that separate optical and acoustic paths, and provides better sensitivity than any optical interferometer. It also provides a non-contact method that is insensitive to optical and acoustic artifacts typical of backward mode of optoacoustic imaging. The LBD sensitivity depends on a large number of system parameters such as probe beam power, spot size, interaction length, optical refraction index of the coupling medium, laser wavelength, photodiode sensitivity, proximity to the optoacoustic source, and thus, can be optimized. The basic setup of OR-LBD-OAM shows high sensitivity competitive with commercial ultrasonic transducers. We report first images of biological cells and tissues obtained using this technique.

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.

A new experimental approach for measurements of temperature dependence of the Gruneisen 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 Gruneisen 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 Gruneisen parameter of DI-water, which we measured by using carbon ink colloid. We also found that Gruneisen-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.

Ultrasound endoscopy has been proven effective in identifying and staging relatively advanced tumors in esophageal or colon wall lining. When combined with optoacoustic imaging modality, the endoscopy examination may prove beneficial in detecting also early stage tumors as well as more accurate staging of advanced tumors based on functional - anatomical maps. Here we present a prototype of a dual-modality optoacoustic – laser ultrasound (OA-LUS) endoscopy system with enhanced imaging capabilities. The system consists of a rotating 90° off-axis parabolic reflector which is acoustically coupled to a flat 8-element transducer array. A parabolic mirror serves dual purpose of directing light towards a sample and reflecting incoming optically generated ultrasound signals towards a detector. LUS modality is enabled by placing an optically absorbing and acoustically transparent polymeric membrane in the path of laser light to generate broadband and non-reverberating transient ultrasound waves propagating towards the sample. Focused system detects ultrasound signals and reconstructs the image similar to optoacoustic mode. Presence of a delay between optically generated and reflected acoustic signals allows concurrent image acquisition in OA and LUS modalities.

Time-resolved piezoelectric detection of wide-band ultrasonic transients induced by laser pulses in absorbing medium was studied. An optoacoustic transducer was developed for measuring the profiles of ultrasonic transients propagating in backward direction out of the laser-irradiated medium. For this purpose, an optical fiber for delivery of laser pulses to the surface of absorbing medium and a wide-band lithium niobate acoustic transducer were incorporated in one compact system, optoacoustic front surface transducer (OAFST). The transducer possesses temporal resolution (rise time) of 3.5 ns, low effective thermal noise pressure (10 Pa), and high sensitivity of piezoelectric detection (10 μV/Pa) over the ultrasonic frequency range from 1 to 100 MHz. Nd:YAG laser pulses at 355 nm were employed to generate distribution of acoustic sources in water solutions of potassium chromate with various concentrations. A temporal course of ultrasonic transients launched into an optically and acoustically transparent me...

Current imaging modalities fail to detect small tumors in the breast. Opto-acoustic tomography is a novel technique for early cancer detection with promising diagnostic capability. The experimental limit of sensitivity and maximal depth of the laser opto-acoustic detection for small model tumors located within bulk phantom tissue were studied. Two phantoms with optical properties similar to that of breast tissue in the near infrared spectral range were used in these studies: turbid gelatin slabs with the thickness of 100 mm and chicken breast muscle slabs with the thickness of up to 80 mm. Gelatin spheres with enhanced absorption coefficient relative to the background absorption and liver tissue were used to simulate small tumors. The experiments demonstrated the capability of laser optoacoustic imaging to detect and localize phantom tumors with the diameter of 2 mm at a depth of up to 60 mm within the gelatin phantoms and 3/spl times/2/spl times/0.6-mm piece of liver tissue within 80-mm chicken breast tissue. Theoretical studies on sensitivity of opto-acoustic detection at various diameters, depths of location, and absorption coefficients of small tumors were performed using the experimental data. Our results suggest that the opto-acoustic imaging may occupy a significant niche in early detection of cancer in the breast and other organs.

A three-dimensional mouse imaging system combining optoacoustic tomography and laser ultrasound (LOUIS-3DM) has been developed. It features broadband laser ultrasound emitters positioned opposite an array of transducers. This imaging geometry allows reconstruction of images that either depicts the speed of sound distribution from measured time of flight data, or acoustic attenuation from the measured signal amplitude. We have investigated the performance of two laser ultrasound source designs, both easily adapta ble to a commercial imaging system: small diameter source (600 µm) generated off a flat surface, and larger diameter (3 mm) spherical source. Laser energy requirements are modest, well below 1 mJ per pulse for either design. Their performance at normal incidence is comparable both in amplitude and frequency response. However, off-axis generation differs dramatically and the shortcomings of th e simple flat emitter design are eviden t. We show that, in order to achieve optimal performance through proper illumination of the detector array, spherical wave front characteristics are desired. Keywords: Broadband ultrasound, laser ultrasonics, wave front propagation, delta-source, spherical wave generation, wide-directivity source. INTRODUCTION We have developed a three-dimensional mouse imaging system (LOUIS-3DM) combining optoacoustic (OA) tomography

We developed the ¯rst prototype of dual-modality imager combining optoacoustic t omography (OAT) and laserultrasound tomography (UST) using computer models followed by experimental va lidation. The system designedfor preclinical biomedical research can concurrently yield images depicting both the abs orbed optical energydensity and acoustic properties (speed of sound) of an object. In our design of the UST imager, we seek toreplace conventional electrical generation of ultrasound waves by laser-induced ultra sound (LU). While earlierstudies yielded encouraging results [Manohar, et al., Appl. Phys. Lett, 131911, 2 007], they were limited totwo-dimensional (2D) geometries. In this work, we conduct computer-simulation st udies to investigate di®erentdesigns for the 3D LU UST imager. The number and location of the laser ultrasound emitters, which areconstrained to reside on the cylindrical surface opposite to the arc of detectors, are optimized. In addition tothe system parameters, an iterative image reconstruction algorithm was o ptimized. We demonstrate that highquality volumetric maps of the speed of sound can be reconstructed when only 32 emitt ers and 128 receivingtransducers are employed to record time-of-°ight data at 360 tomographic view ang les. The implications of theproposed system for small animal and breast-cancer imaging are discussed.Keywords: Laser ultrasound tomography, optoacoustic tomography, iterative ima ge reconstruction

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.

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.).

Alexandrite laser ablation of enamel enhanced by an indocyanine green dye was studied. A microjet system was employed to deliver precisely measured small amounts of absorbing dye solution to the site of irradiation. A sequence of physical phenomena involved in dye-enhanced laser ablation of dental enamel was revealed when laser pulse profiles were compared with the profiles of laser-induced pressure, laser-induced plasma, and ablation plume kinetics. To understand photomechanical effects on the enamel ablation, the absolute values of pressure waves were measured by a calibrated wide-band acoustic transducer. Absolute amplitude and temporal profile of pressure waves, plasma emission, ablation plume kinetics, ablation efficiency, and crater quality under free-running and and Q-switched ablation of enamel were studied. It was found that there is an optimal dye solution volume (100-200 nL) when the maximum ablation efficiency (30 μm/pulse) can be obtained. It was shown that the ablation efficiency under Q-switched laser irradiation is approximately one order of magnitude lower than that under free-running ablation. It was shown that Q-switched enamel ablation with dye solution is caused by the powerful recoil pressure wave with an amplitude 3-6.5 kbar. In contrast, dye-enhanced free-running enamel ablation is caused by plasma-mediated evaporation of enamel and accompanied by recoil pressure waves of lower amplitude (0.5-1 kbar) that is below mechanical damage threshold in enamel. Uneven crater walls after Q-switched ablation were observed by scanning electron microscopy (SEM). Free-running ablation makes precise craters with smooth and even crater walls.

Background and Objectiue: It is desirable for laser microsurgical procedures to remove tissue accurately and with minimal thermal and mechanical damage to adjacent non-irradiated tissues. Pulsed laser ablation can potentially remove biological tissue with microprecision if appropriate irradiation conditions are applied. The major goal of this study was to determine whether laser ablation is possible at temperatures below 100°C. Another aim was to test thermoelastic and recoil stress magnitudes and to estimate their effects on phantom and biological tissue. Study DesignlMateriaZs and Methods: Pulsed laser ablation of water (aqueous solution of potassium chromate) and water containing soft tissues (collagen gel and pig liver) irradiated under confined stress conditions was studied. The ablation mechanism and stages of the ablation process were determined based on time-resolved measurements of laser-induced acoustic waves with simultaneous imaging of the ablation process by laser-flash photography. Results: This study reveals the important role of tensile thermoelastic stress, which produces efficient cavitation that drives material ejection at temperatures substantially below 100°C. Ablation thresholds for the aqueous solution, collagen gel, and liver were 20,38, and 55 J/cm”, respectively, which correspond to temperature jumps of 5,10, and 15°C. Two distinct stages of material ejection were observed: (1) initial removal of small volumes of material due to the rupture of single subsurface bubbles, (2) bulk material ablation in the form of jets produced by intense hydrodynamic motions formed upon collapse of large bubbles after coalescence of smaller bubbles. The duration of material ejection upon short-pulse ablation may vary from microseconds to submilliseconds, and depended on the mechanical properties of materials and the incident laser fluence. ConcZusion: Nanosecond laser ablation of water, gels, and soft tissue under confined-stress conditions of irradiation may occur at temperatures below 100°C. This ablation regime minimizes thermal injury to adjacent tissues and involves thermoelastic stress and recoil pressure magnitudes, which may be tolerated by tissues adjacent to an ablated crater. Q 1996 Wiley-Liss, Inc.

Plasma mediated ablation of collagen gels and porcine cornea was studied at various laser pulse durations in the range of 1 ns-300 fs at 1053-nm wavelength. It was found that pulsed laser ablation of transparent and weakly absorbing gels is always mediated by plasma. On the other hand, ablation of strongly absorbing tissues is mediated by plasma in the ultrashort-pulse range only. Ablation threshold along with plasma optical breakdown threshold decreases with increasing tissue absorbance for subnanosecond pulses. In contrast, the ablation threshold was found to be practically independent of tissue linear absorption for femtosecond laser pulses. The mechanism of optical breakdown at the tissue surface was theoretically investigated. In the nanosecond range of laser pulse duration, optical breakdown proceeds via avalanche ionization initiated by heating of electrons contributed by strongly absorbing impurities at the tissue surface. In the ultrashortpulse range, optical breakdown is initiated by multiphoton ionization of the irradiated medium (six photons in case of tissue irradiated at 1053-nm wavelength), and is less sensitive to linear absorption. High-quality ablation craters with no thermal or mechanical damage to surrounding material were obtained with subpicosecond laser pulses. Experimental results suggest that subpicosecond plasma mediated ablation can be employed as a tool for precise laser microsurgery of various tissues.

This paper provides a short review of physical principles, technology, biomedical applications and perspectives of the optoacoustic imaging. Ideas that made this rapidly developing field possible include the following: (1) laser pulses may be effectively used to produce acoustic pressure in biological tissues localized to the areas of increased optical absorption, (2) the resulting acoustic (ultrasonic) waves propagate in tissues with minimal distortions and attenuation, and (3) 2D and 3D maps (images) of the absorbed optical energy can be reconstructed with high resolution from the time-resolved optoacoustic signals. Modern optoacoustic imaging systems include scanning focused transducers and transducer arrays. The widely accepted linear arrays are employed either for real-time 2D optoacoustic imaging or for 3D imaging via translational or rotational scanning. A commercial prototype of a 3D OAT system was developed by TomoWave Labs for applications in preclinical research, where major biomedical applications include visualization of specific targeting using exogenous optoacoustic contrast agents and imaging of blood distribution and oxygentaion status.

In the present work, we demonstrate a potential use of gold nanorods as a contrast agent for selective photothermal therapy of human acute leukemia cells (HL-60) using a near-infrared laser. Gold Nanorods (GNR) are synthesized and conjugated to CD33, a 67 kDa glycoprotein found on the surface of myeloid cells that belongs to the sialoadhesin family of proteins. After pegylation, or conjugation with CD33 antibody, GNR were non-toxic for acute and chronic leukemia cells. We used a Quanta System q-switched titanium sapphire laser emitting at a center wavelength of 755 nm. Each sample was illuminated with 1 laser shot at either high or low fluence. Both laser modes were used in 3 independent cell probes. HL-60 cells were treated for 45 min with GNR conjugated with mAb CD33, or with GNR-Pegylated particles. After laser application, the cells were resuspended and analyzed to cell viability with Trypan blue exclusion assay. GNR-CD33 conjugates significantly increase the percentage of cell death as compared with a control group after laser illumination: a 3 fold increase is observed.

XeCl laser ablation of atherosclerotic aorta tissue was investigated. Luminescence spectra of ablation products were measured for soft and hard arterial tissues. A pronounced difference observed between plume luminescence for various plaques and normal vessel wall correlates with the chemical composition of the tissue. The mechanism of plume luminescence appeared to be thermochemical excitation of ablation products (particles, atoms, molecules, etc.) in the air. The process of soft tissue ablation was delayed with respect to the exciting laser pulse at relatively low laser fluences close to the ablation threshold. The kinetics of the ablation process as a function of laser-pulse energy fluence is reported. The data indicate that tissue ejection mechanism, which involves vapor bubbles formation, expansion and explosion, is suitable for the description of the XeCl excimer laser ablation of soft tissues. © 1993 Wiley-Liss, Inc.

Quantitative studies are conducted into the absolute pressure values of the acoustical and shock waves generated and propagating in a biotissue under pulsed (tau p = 50 ns) UV (lambda = 308 nm) laser irradiation (below and above the ablation threshold). Powerful (several hundreds of bars in pressure) high-frequency (f approximately 10(7) Hz) acoustic compression and rarefaction pulses are found to be generated in the biotissue. The amplitudes and profiles of the acoustic pulses developing in atherosclerotic human aorta tissues and an aqueous CuCl2 solution under laser irradiation are investigated as a function of the laser pulse energy fluence. The results obtained point to the absence of the cold spallation of the objects of study by rarefaction waves. Based on experimental data, the rise rates, pressure gradients, and propagation velocities of shock waves in the biotissue are calculated. The experimental data are found to agree well with the theoretical estimates.

We present a non-metallic interferometric silica optical fiber ultrasonic wideband sensor for optoacoustic imaging applications. The ultrasonic sensitivity of this sensor has been characterized over the frequency range from 1 to 10 MHz. A comparative analysis has been carried out between this sensor and an array of piezoelectric transducers using optoacoustic signals generated from an optical absorbent embedded in a tissue mimicking phantom. Also, a two dimensional reconstructed image of the phantom using the fiber interferometric sensor is presented and compared to the image obtained using the Laser Optoacoustic Imaging System, LOIS-64B. The feasibility of our fiber optic based sensor for wideband ultrasonic detection is demonstrated. (© 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

A 3-D optoacoustic imaging system was used to visualize thermal lesions produced in vivo using high-intensity focused ultra- sound 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. Tomog- raphic 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 recon- struct 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 le- sion 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 pre- liminary results demonstrate the potential of optoacoustic imaging to assess and monitor the progress of HIFU therapy. © 2010 Society of Photo

A non-metallic interferometric silica optical fiber ultrasonic wideband sensor is presented for optoacustic imaging applications. We characterized the ultrasonic sensitivity of the optical fiber sensor within the frequency range of 1 to 5 MHz. We compared detection of real world optoacoustic signals, generated from an optically absorbing object embedded in a tissue mimicking phantom, between our sensor and an array of piezoelectric transducers. Reconstructed two dimensional acoustic images of the phantom are presented and compared with images obtained with the Laser Optoacoustic Imaging System, LOIS-64B, demonstrating the feasibility of our fiber optic sensor as a wideband ultrasonic sensor.

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.

We have investigated the limitations of our laser ultrasonic plane wave δ-source. In theory, the device is capable of producing an acoustic impulse with a bandwidth exceeding 30 MHz. However, a bandwidth of 12 MHz is measured with a calibrated wideband hydrophone. A test setup was designed and built. It permits the investigation of experimental parameters that alter the generated acoustic impulse: laser pulse duration, laser spatial profile, and absorber opacity. Laser energy spatial profile is the main contributor to the narrowing of the frequency band. Our findings are presented, along with further justifications for a device with very large effective area.

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.

The energetics of 308-nm excimer laser irradiation of human aorta were studied. The heat generation that occurred during laser irradiation of atherosclerotic aorta equaled the absorbed laser energy minus the fraction of energy for escaping fluorescence (0.8-1.6%) and photochemical decomposition (2%). The absorbed laser energy is equal to the total delivered light energy minus the energy lost as specular reflectance (2.4%, air/tissue) and diffuse reflectance (11.5-15.5%). Overall, about 79-83.5% of the delivered light energy was converted to heat. We conclude that the mechanism of XeCl laser ablation of soft tissue involves thermal overheating of the irradiated volume with subsequent explosive vaporization. The optical properties of normal wall of human aorta and fibrous plaque, both native and denatured were determined. The light scattering was significant and sufficient to cause a subsurface fluence (J/cm2) in native aorta that equaled 1.8 times the broad-beam radiant exposure, phi o (2.7 phi o for denatured aorta). An optical fiber must have a diameter of at least 800 microns to achieve a maximum light penetration (approximately 200 microns for phi o/e) in the aorta along the central axis of the beam.

An interferometric sensor based on nonmetallic silica optical fiber is presented as an ultrasonic wideband transducer for optoacoustic imaging applications. We have characterized the sensitivity of the optical fiber sensor by detecting optoacoustic signals from an optically absorbing object embedded in a tissue-mimicking phantom and have compared the signals recorded with those detected from the same phantom using an array of piezoelectric transducers. The optical fiber sensor was also scanned along the phantom surface in order to reconstruct two-dimensional optoacoustic images of the phantom. These images have been compared with images obtained using the Laser Optoacoustic Imaging System, LOIS-64B, demonstrating the feasibility of our fiber-optic sensor as a wideband ultrasonic transducer.

Comprehensive characterization of wideband ultrasonic transducers and specifically optoacoustic detectors is achieved through the analysis of their frequency response as a function of the incident angle. The tests are performed under well-defined, repeatable operating conditions. Backillumination of a blackened, acoustically matched planar surface with a short laser pulse creates an acoustic impulse which is used as a wideband ultrasonic source. Upon illumination with a short laser pulse, the bandwidth of our source shows a −6 dB point of 12 MHz and a low-frequency roll-off around 300 kHz. Using proprietary software, we examine thoroughly the planarity of the emitted wave front within a specified amplitude cutoff and phase incoherence. Analysis of the angular dependence of the frequency response yields invaluable directivity information about the detector under study: a necessary component toward accurate optoacoustic image reconstruction and quantitative tomography. The laser ultrasonic source we developed is the main feature of our directivity measurement setup. Due to its simplicity, it can easily be adapted to various calibration devices. This paper focuses on the development and characterization of the flatness and the bandwidth of our wideband ultrasonic source.

We designed, fabricated and tested the laser optoacoustic imaging system for breast cancer detection LOIS-64, which fuses optical and acoustic imaging techniques in one modality by utilizing pulsed optical illumination and ultrawide-band ultrasonic detection of resulting optoacoustic OA signals. The system was designed to im- age a single breast slice in craniocaudal or mediolateral projection with an arc-shaped array of 64 ultrawide-band acoustic transducers. The system resolution on breast phantoms was at least 0.5 mm. The single-channel sensitivity of 1.66 mV/Pa was estimated to be suffi- cient for single-pulse imaging of 6t o 11 mm tumors through the whole imaging slice of the breast. The implemented signal processing using the wavelet transform allowed significant reduction of the low- frequency LF acoustic noise, allowed localization of the optoacous- tic signals from tumors, and enhanced the contrast and sharpened the boundaries of the optoacoustic images of the tumors. During the pre- liminary clinical studies on 27 patients, the LOIS-64 was able to visu- alize 18 out of 20 malignant lesions suspected from mammography and ultrasound images and confirmed by the biopsy performed after the optoacoustic tomography OAT procedure. © 2009 Society of Photo

We present a physical model that explains several sequential stages of the conversion of optical to acoustical energy when irradiating diluted suspensions of metal nanoparticles with laser pulses. Optical absorption and scattering of a single particle driven by plasmon resonance interactions in an aqueous medium are considered. Thermal effects produced by laser-irradiated nanoparticles, dynamics of vapor bubble formation, and acoustic signals from expanding bubbles formed around heated nanoparticles are calculated. Stochastic features of the pressure magnitude emitted as a result of low-fluence irradiation of suspensions are also discussed. The probabilistic distribution of pressure magnitude from individual bubbles was found to obey Zipf's law for low concentrations of nanoparticles, while increasing their concentration brings the pressure magnitude distribution into conformance with the Gaussian law.

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 vess els, respectively. Filtering of the optoacoustic signals using high frequency enhancing wavelets further emphasized the smaller blood vessels. Keywords : small animal imaging, optoacoustics, image reconstruction, functional imaging, blood, vasculature.

We demonstrated the ability to detect surface antigens, associated with pathogens, utilizing laser optoacoustic spectroscopy with antibody-coupled gold nanorods (GNR) as a contrast agent specifically targeted to the antigen of interest. The sensitivity of the technique has been assessed by determining the minimum detectable concentration of a surface antigen from biological samples. We compared the sensitivity and applicability of two different methods for detecting optoacoustic responses, using e ither optical or piezoelectric sensors. Bi ological samples were adsorbed to the inside walls of detachable, flat-bottomed plastic micro-wells, and then probed with appropriate antibodies conjugated with gold nanorods. If the target antigens were present, the antibody-nanoparticle conjugates were bound, while any nonadsorbed nanoparticles were washed out of the wells. Optoacoustic signals were generated from the bound nanorods using a pulsed pump laser at wavelengths corresponding to one of the peak absorptions of the nanorods. Optoacoustic responses were obtained from the samples using both detection modalities. The sensitivity, suitability, ease of use of each method were assessed and compared. Initial results indicat e that optical detection gives comparable sensitivity as the piezoelectric method, and further enhancement of the detection sensitivity is possible for both methods. An advantage of the piezoelectric de tection method is that it may be implemente d in a more compact assembly, compared to the optical method, however, the optical method may be less sensitive to external electromagnetic and acoustic noise. Further evaluation will be required to refine these measurements . The results with both met hods indicate that the use of antibody-targeted nanorod contrast agents, with laser-optoacoustic detection, is a promising technology for the development of rapid in vitro diagnostic tests. Keywords : pathogenic detection, optoacoustic, gold nanoparticles, piezoelectric, probe beam deflection technique 1. INTRODUCTION Combination of optoacoustic detection methods, known for their high sensitivity, with contrast agents based on metal nanoparticles that possess strong optical absorption and effective conversion of the optical energy into heat promises to bring about a large number of biomedical applications [1]. These applications include imaging [2], sensing [3] and monitoring of targeted therapy [4, 5]. Optoacoustic detection of very small concentration of cells, bacteria or other micro-organisms may open doors for an effective early diagnostics of deceases [6, 7]. An essential aspect of the application of optoacoustic techniques to biomedical problems is the optimization of the methods used for recording the acoustic responses induced in the biological target, such as pathological microorganism (PMO). Most researchers in this field utilize acoustically coupled piezoelectric transducers (ultrasonic hydrophones) for this purpose, directly detecting laser-induced pressure transients. It is also possible to detect these signals optically, exploiting either (etalon) interferometry [8] or a technique ba sed on the change in refractive index induced in the sample medium due to propagation of the pressure waves [7, 9-11]. In the present study, the technical aspects of detecting optoacoustic signals by an ultrawide-band ultrasonic transdu cer were compared with the tr ansient pressure measurement using optical beam deflection method. The ultimate goal of our study is to develop an optoacoustic biosensor for blood

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.

A three-dimensional laser optoacoustic imaging system was developed, which combines the advantages of optical spectroscopy and high resolution ultrasonic detection, to produce high contrast maps of optical absorbance in tissues. This system was tested in a nude mouse model of breast cancer and produced tissue images of tumors and vasculature. The imaging can utilize either optical properties of hemoglobin and oxyhemoglobin, which are the main endogenous tissue chromophores in the red and near-infrared spectral ranges, or exogenous contrast agent based on gold nanorods. Visualization of tissue molecules targeted by the contrast agent provides molecular information. Visulization of blood at multiple colors of light provides functional information on blood concentration and oxygen saturation. Optoacoustic imaging, using two or more laser illumination wavelengths, permits an assessment of the angiogenesis-related microvasculature, and thereby, an evaluation of the tumor stage and its metastatic potential. The optoacoustic imaging system was also used to generate molecular images of the malignancy-related receptors induced by the xenografts of BT474 mammary adenocarcinoma cells in nude mice. The development of the latter images was facilitated by the use of an optoacoustic contrast agent that utilizes gold nanorods conjugated to monoclonal antibody raised against HER2/neu antigens. These nanorods possess a very strong optical absorption peak that can be tuned in the near-infrared by changing their aspect ratio. The effective conversion of the optical energy into heat by the gold nanorods, followed by the thermal expansion of the surrounding water, makes these nanoparticles an effective optoacoustic contrast agent. Optical scattering methods and x-ray tomography may also benefit from the application of this contrast agent. Administration of the gold nanorod bioconjugates to mice resulted in an enhanced contrast of breast tumors relative the background of normal tissues in the nude mouse model. The combination of this novel contrast agent and optoacoustic imaging has the potential to become a useful imaging modality, for preclinical research in murine models of cancer and other human diseases.