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We recently built a fiber-optic laser-ultrasound (LU) scanner for nondestructive evaluation (NDE) of aircraft composites and demonstrated its greatly improved sensitivity and stability compared with current noncontact systems. It is also very attractive in terms of cost, stability to environmental noise and surface roughness, simplicity in adjustment, footprint, and flexibility. A new type of a balanced fiber-optic Sagnac interferometer is a key component of this all-optical LU pump-probe system. Very high A-scan rates can be achieved because no reference arm or stabilization feedback are needed. Here, we demonstrate LU system performance at 1000 A-scans/s combined with a fast 2-D translator operating at a scanning speed of 100 mm/s with a peak acceleration of 10 m/s(2) in both lateral directions to produce parallel B-scans at high rates. The fast scanning strategy is described in detail. The sensitivity of this system, in terms of noise equivalent pressure, was further improved to be only 8.3 dB above the Nyquist thermal noise limit. To our knowledge, this is the best reported sensitivity for a noncontact ultrasonic detector of this dimension used to inspect aircraft composites.

The advantages of photoacoustic imaging, including low cost, non-ionizing operation, and sub-mm spatial resolution at centimeters depth, make it a promising modality to probe nanoparticle-targeted abnormalities in real time at cellular and molecular levels. However, detecting rare cell types in a heterogeneous background with strong optical scattering and absorption remains a big challenge. For example, differentiating circulating tumor cells in vivo (typically fewer than 10 cells/ml for an active tumor) among billions of erythrocytes in the blood is nearly impossible. In this paper, a newly developed technique, magnetomotive photoacoustic (mmPA) imaging, which can greatly increase the sensitivity and specificity of sensing targeted cells or molecular interactions, is reviewed. Its primary advantage is suppression of background signals through magnetic enrichment/manipulation with simultaneous photoacoustic detection of magnetic contrast agent targeted objects. Results from phantom and in vitro studies demonstrate the capability of mmPA imaging to differentiate regions targeted with magnetic nanoparticles from the background, and to trap and sensitively detect targeted cells at a concentration of a single cell per milliliter in a flow system mimicking a human peripheral artery. This technique provides an example of the ways in which molecular imaging can potentially enable robust molecular diagnosis and treatment, and accelerate the translation of molecular medicine into the clinic.

Circulating tumor cells (CTCs) have been reported to correlate most closely with cancer development, and can serve as an important marker for metastatic malignancy, tumor recurrence, and prediction of prognosis and therapeutic effi cacy. Detecting and quantifying CTCs, however, have proven to be challenging due to their low abundance in blood. Based on magneto-optical coupled nanoprobes (made of gold nanorod and iron oxide nanoparticles) and photoacoustic (PA) imaging, we report the development of an enabling technology that can detect CTCs at single cell/mL level. Remarkably, at this low cell concentration, approximately 67% of circulating tumor cells can be captured and imaged with just one pass through the magnetic trapping zone. Compared to the conventional in vitro assays, this technology offers signifi cantly improved sensitivity, because it is inherently compatible with large sample volumes. Compared to more advanced in vivo CTC detection methods, this technology can solve the low throughput problem of optical imaging, and streamline the photoacoustic imaging process by combining magnetic enrichment and digital readout into a single step. Most cancer deaths are caused by metastasis, a process

Photoacoustic (PA) imaging has been used mainly for anatomical and functional imaging. Although functionalized nanoparticles also have been developed for PA molecular imaging, only single targeting has been demonstrated. In this study, PA imaging of multiple targets using gold nanorods is demonstrated experimentally using HER2 and CXCR4 as target molecules. The two corresponding monoclonal antibodies were conjugated to two types of gold nanorod with different aspect ratios. Gold nanorods with mean aspect ratios of 5.9 and 3.7 exhibited peak optical absorptions at 1000 and 785 nm, respectively. Appropriate selection of laser irradiation wavelength enhances PA signals by 7-12 dB and allows signals from gold nanorods corresponding to specific bindings to be distinguished. This approach potentially allows the expression levels of different oncogenes of cancer cells to be revealed simultaneously.

To integrate real-time photoacoustics (PA) into ultrasound (US) scanners and accelerate clinical translation of combined PAUS imaging, we previously developed a system using a portable, low-cost, low pulse energy, high-repetition rate laser (~1kHz) with a 1D galvo-mirror for rapid laser beam scanning over the imaging area. However, the frame rate and pulse energy are limited because of regulations on the radiance (1 W/cm(2)). Therefore, a laser scan scheme needs to be optimized to provide high frame rate within this safety limit. In addition, the laser light should be evenly distributed to minimize any artifacts caused by the scanning approach. In this paper, we calculated the laser light distribution using 3D Monte Carlo simulation and further developed the system to scan the laser beam in elevation as well as laterally using a 2-dimensional galvo-mirror scanner to achieve higher frame rates within the radiance safety limit. Insertion of a needle into chicken breast tissue was used to demonstrate our optimized scan scheme.

Ultrasound-guided photoacoustic imaging has shown great potential for many clinical applications including vascular visualization, detection of nanoprobes sensing molecular profiles, and guidance of interventional procedures. However, bulky and costly lasers are usually required to provide sufficient pulse energies for deep imaging. The low pulse repetition rate also limits potential real-time applications of integrated photoacoustic/ultrasound (PAUS) imaging. With a compact and low-cost laser operating at a kHz repetition rate, we aim to integrate photoacoustics (PA) into a commercial ultrasound (US) machine utilizing an interleaved scanning approach for clinical translation, with imaging depth up to a few centimeters and frame rates > 30 Hz. Multiple PA sub-frames are formed by scanning laser firings covering a large scan region with a rotating galvo mirror, and then combined into a final frame. Ultrasound pulse-echo beams are interleaved between laser firings/PA receives. The approach was implemented with a diode-pumped laser, a commercial US scanner, and a linear array transducer. Insertion of an 18-gauge needle into a piece of chicken tissue, with subsequent injection of an absorptive agent into the tissue, was imaged with an integrated PAUS frame rate of 30 Hz, covering a 2.8 cm × 2.8 cm imaging plane. Given this real-time image rate and high contrast (> 40 dB at more than 1-cm depth in the PA image), we have demonstrated that this approach is potentially attractive for clinical procedure guidance.

Phase transition contrast agents were first introduced in ultrasound (US) in the form of perfluorocarbon droplets. When their size is reduced to the nanoscale, surface tension dominates their stability and high pressure is required to vaporize them using long US emissions at high frequencies. Our group recently showed that nanoemulsion beads (100-300 nm) coated with gold nanopsheres could be used as non-linear contrast agents. Beads can be vaporized with light only, inducing stronger photoacoustic signals by increasing thermal expansion. A photoacoustic cavitation threshold study (US: 1.2 MHz, Laser 750 nm and 10-ns pulse) shows that the vaporization thresholds of NEB-GNS can be greatly reduced using simultaneous light and US excitations. The resulting signal is driven only by the pressure amplitude for a fluence higher than 2.4 mJ/cm2. At diagnostic exposures, it is possible to capture very high signals from the vaporized beads at concentrations reduced to 10 pM with optical absorption smaller than 0.01 cm-1. A real-time imaging mode selectively isolating vaporization signals was implemented on a Verasonics system. A linear US probe (L74, 3 MHz) launched short US bursts before light was emitted from the laser. Vaporization of NEB-GNS resulted in a persistent 30-dB signal enhancement compared to a dye with the same absorption. Specific vaporization signals were retrieved in phantom experiments with US scatterers. This technique, called sonophotoacoustics, has great potential for targeted molecular imaging and therapy using compact nanoprobes with potentially high-penetrability into tissue.

Photoacoustic imaging has emerged as a highly promising tool to visualize molecular events with deep tissue penetration. Like most other modalities, however, image contrast under in vivo conditions is far from optimal due to background signals from tissue. Using iron oxide–gold core–shell nanoparticles, we have previously demonstrated the concept of magnetomotive photoacoustic (mmPA) imaging, which is capable of dramatically reducing the influence of background signals and producing high-contrast molecular images. Here, we report two significant advances toward clinical translation of this technology. First, we introduce a new class of compact, uniform, magneto-optically coupled core–shell nanoparticles, prepared through localized copolymerization of polypyrrole (PPy) on an iron oxide nanoparticle surface. The resulting iron oxide–PPy nanoparticles feature high colloidal stability and solve the photoinstability and small-scale synthesis problems previously encountered by the gold coating approach. In parallel, we have developed a new generation of mmPA featuring cyclic magnetic motion and ultrasound speckle tracking (USST), whose imaging capture frame rate is several hundred times faster than the photoacoustic speckle tracking (PAST) method we demonstrated previously. These advances enable robust artifact elimination caused by physiologic motions and demonstrate the application of the mmPA technology for in vivo sensitive tumor imaging.

Because of depth-dependent light attenuation, bulky, low-repetition-rate lasers are usually used in most photoacoustic (PA) systems to provide sufficient pulse energies to image at depth within the body. However, integrating these lasers with real-time clinical ultrasound (US) scanners has been problematic because of their size and cost. In this paper, an integrated PA/US (PAUS) imaging system is presented operating at frame rates >30 Hz. By employing a portable, low-cost, low-pulse-energy (~2 mJ/pulse), high-repetition-rate (~1 kHz), 1053-nm laser, and a rotating galvo-mirror system enabling rapid laser beam scanning over the imaging area, the approach is demonstrated for potential applications requiring a few centimeters of penetration. In particular, we demonstrate here real-time (30 Hz frame rate) imaging (by combining multiple single-shot sub-images covering the scan region) of an 18-gauge needle inserted into a piece of chicken breast with subsequent delivery of an absorptive agent at more than 1-cm depth to mimic PAUS guidance of an interventional procedure. A signal-to-noise ratio of more than 35 dB is obtained for the needle in an imaging area 2.8 × 2.8 cm (depth × lateral). Higher frame rate operation is envisioned with an optimized scanning scheme.

Laser ultrasonic (LU) inspection represents an attractive, non-contact method to evaluate composite materials. Current non-contact systems, however, have relatively low sensitivity compared to contact piezoelectric detection. They are also difficult to adjust, very expensive, and strongly influenced by environmental noise. Here, we demonstrate that most of these drawbacks can be eliminated by combining a new generation of compact, inexpensive fiber lasers with new developments in fiber telecommunication optics and an optimally designed balanced probe scheme. In particular, a new type of a balanced fiber-optic Sagnac interferometer is presented as part of an all-optical LU pump-probe system for high speed non-destructive testing and evaluation (NDT&E) of aircraft composites. The performance of the LU system is demonstrated on a composite sample typically used in the aircraft industry. Wide-band ultrasound probe signals are generated directly at the sample surface with a pulsed diode-pumped laser delivering nanosecond laser pulses at a 1 kHz repetition rate with a pulse energy of 2 mJ. A balanced fiber-optic Sagnac interferometer is employed to detect pressure signals in a 1-10 MHz frequency range at the same point (an 8 μm focal spot) on the composite surface. A fast (up to 100 mm/s) 2D translation system is employed to move the sample during scanning and produce a complete B-scan consisting of one thousand A-scans in less than a second. The sensitivity of this system, in terms of the noise equivalent pressure, is found to be only 10 dB above the Nyquist thermal noise limit. To our knowledge, this is the best reported sensitivity for a non-contact ultrasonic detector of this dimension.

Ultrasound (US) needle tracking with a real-time, portable imaging system is common for biopsies and therapeutic injections. However, proper US probe-needle orientation is required or tracking precision is lost because of poor US contrast and additional artifacts. In contrast, the strong photoacoustic (PA) signal from a needle is relatively independent of light orientation. Unfortunately, a bulky, slow (10s of Hz) laser is usually required for sufficient pulse energy at centimeters depth inside the body, hindering PA integration into a real-time US scanner. We employ a compact, low-cost, low pulse energy (1 mJ/pulse) laser that can operate at 10s of kHz to construct an integrated US/PA system with frame rates higher than 30 Hz. A scanning approach with a rotating galvo mirror was used by combining multiple laser shots covering a large scan region to form an integrated image. Multiple pulse-echo US focused beams were formed between laser firings. Insertion of an 18-gauge needle in a piece of chicken breast tissue, and subsequent injection of an absorptive agent through the needle, was imaged. The PA image displays the needle at higher contrast (> 30 dB) than the US image, even at a depth of 12 mm where light energy has largely decayed, demonstrating the potential of the current scanning system for real-time needle guidance. An improved frame rate is envisioned with an optimized scan scheme.

Optically activated cavitation in a nanoemulsion contrast agent is proposed for therapeutic applications. With a 56°C boiling point perfluorohexane core and highly absorptive gold nanospheres at the oil–water interface, cavitation nuclei in the core can be efficiently induced with a laser fluence below medical safety limits (70 mJ/cm(2) at 1064 nm). This agent is also sensitive to ultrasound (US) exposure and can induce inertial cavitation at a pressure within the medical diagnostic range. Images from a high-speed camera demonstrate bubble formation in these nanoemulsions. The potential of using this contrast agent for blood clot disruption is demonstrated in an in vitro study. The possibility of simultaneous laser and US excitation to reduce the cavitation threshold for therapeutic applications is also discussed.

Ultrasound-induced inertial cavitation is a mechanical process used for site-localized therapies such as non-invasive surgery. Initiating cavitation in tissue requires very high intensity focused ultrasound (HIFU) and low-frequencies. Hence, some applications like thrombolysis require targeted contrast agents to reduce peak intensities and the potential for secondary effects. A new type of theranostic nanoemulsion has been developed as a combined ultrasound (US)/photoacoustic(PA) agent for molecular imaging and therapy. It includes a nanoscale emulsion core encapsulated with a layer of gold nanospheres at the water/ oil interface. Its optical absorption exhibits a spectrum broadened up to 1100 nm, opening the possibility that 1064 nm light can excite cavitation nuclei. If optically-excited nuclei are produced at the same time that a low-frequency US wave is at peak negative pressure, then highly localized therapies based on acoustic cavitation may be enabled at very low US pressures. We have demonstrated this concept using a low-cost, low energy, portable 1064 nm fiber laser in conjunction with a 1.24 MHz US transducer for simultaneous laser/US excitation of nanoemulsions. Active cavitation detection from backscattered signals indicated that cavitation can be initiated at very low acoustic pressures (less than 1 MPa) when laser excitation coincides with the rarefaction phase of the acoustic wave, and that no cavitation is produced when light is delivered during the compressive phase. US can sustain cavitation activity during long acoustic bursts and stimulate diffusion of the emulsion, thus increasing treatment speed. An in vitro clot model has been used to demonstrate combined US and laser excitation of the nanoemulsion for efficient thrombolysis.

Due to the high scattering coefficient of tissue over the wavelength range used for photoacoustic (PA) imaging, most studies employ bulky, low repetition rate lasers to provide sufficient pulse energies at depth to image within the body. The size and cost of these lasers has impeded integration of photoacoustics into conventional, routinely-used ultrasound (US) scanners. Here, we present an approach leveraging the capabilities of modern, high repetition rate fiber lasers to produce a clinically translatable system providing integrated US/PA images at frame rates > 30 Hz. The system uses a portable, low-cost, low pulse-energy (1 mJ/pulse), high repetition rate (1 kHz), 1064 nm laser and is designed for integrated US/PA imaging of the peripheral vasculature or any relevant diseased region, such as a tumor. Using a rotating galvo-mirror system, the incident laser beam is quickly scanned over the imaging area. Multiple PA images covering the scan area are integrated to form a single PA image. Additionally, ultrasound firings are integrated into the scan sequence to provide an US image reconstructed over the same frame period. We acquired PA images of a 1.5-mmdiameter cylindrical absorber (absorption coefficient 5 cm -1 ) embedded in a tissue-mimicking gelatin phantom at 6-cm depth. A 2 cm × 1 cm (depth × lateral) area was reconstructed. We obtained a signal-to-noise ratio of more than 30 dB, comparable to conventional PA methods using high energy, low repetition rate lasers. The current system produces an integrated US/PA frame at a 32 Hz rate, and 100 Hz frame rates are possible with our present approach.

A nanoemulsion contrast agent with a perfluorohexane core and optically absorptive gold nanospheres (GNSs) assembled on the surface, is presented to improve the specificity of photoacoustic (PA) molecular imaging in differentiating targeted cells or aberrant regions from heterogeneous background signals. Compared to distributed GNSs, clustered GNSs at the emulsion oil-water interface produce a red-shifted and broadened absorption spectrum, exhibiting fairly high absorption in the near-infrared region commonly used for deep tissue imaging. Above a certain laser irradiation fluence threshold, a phase transition creating a microbubble in the emulsion core leads to more than 10 times stronger PA signals compared with conventional thermal-expansion-induced PA signals. These signals are also strongly non-linear, as verified by a differential scheme using recorded PA images at different laser fluences. Assuming a linear relation between laser fluence and the PA signal amplitude, differential processing results in nearly perfect suppression of linear sources, but retains a significant residue for the non-linear nanoemulsion with more than 35 dB enhancement. This result demonstrates that contrast specificity can be improved using the nanoemulsion as a targeting agent in PA molecular imaging by suppressing all background signals related to a linear PA response. Furthermore, combined with a system providing simultaneous laser/ultrasound excitation, cavitation-generated bubbles have the potential to be a highly specific contrast agent for ultrasound molecular imaging and harmonic imaging, as well as a targeted means for noninvasive ultrasound-based therapies.

A new technique using a composite contrast agent is proposed to increase specific contrast in photoacoustic (PA) molecular imaging. The composite particle has an emulsion bead core with a surface-assembled monolayer of gold nanospheres (GNSs), which efficiently absorbs optical energy under laser irradiation and promotes a phase transition of the emulsion bead to form a transient bubble. Much stronger PA signals are generated by the phase transition process than those from thermal expansion, the most common mechanism of laser-sound conversion. By applying a differential scheme of images, recorded at a series of variable fluences, assuming a linear relation between laser fluence and the PA signal amplitude, a nearly perfect suppression of a linear agent can be obtained, while large residues for the nonlinear emulsion beads remain. This result clearly indicates enhanced PA sensitivity and specificity. Localized bubble generation activated by a short laser pulse was also demonstrated using a microscope coupled with a high-speed camera. With simultaneous ultrasound/laser probing, i.e., delivering a probe ultrasound beam after bubble formation, the nonlinear agent shows great potential for ultrasound contrast-enhanced imaging and harmonic imaging.

The possibility of manipulating inertial cavitation induced in a new type of nanoemulsion contrast agent with simultaneous nanosecond pulsed laser and ultrasound illumination is described. The contrast agent consists of an encapsulated emulsion core coated with a layer of 12nm-diameter gold nanospheres. A low-cost, high repetition-rate, low-energy 1064 nm fiber laser transiently heated the emulsion beads to help initiate inertial cavitation due to a phase transition in the emulsion core. It has been shown that inertial cavitation can be initiated at a very low acoustic pressure (0.43MPa) with laser irradiation applied at the rarefaction phase of the incident acoustic wave. The significantly decreased inertial cavitation threshold in this nanoemulsion suggests that it could be an effective tool for site-targeted, molecular therapeutics in addition to its proposed use as a highly specific molecular imaging agent for photoacoustics.

Photoacoustic (PA) imaging has been widely used in molecular imaging to detect diseased cells by targeting them with nanoparticle-based contrast agents. However, the sensitivity and specificity are easily degraded because contrast agent signals can be masked by the background. Magnetomotive photoacoustic imaging uses a new type of multifunctional composite particle combining an optically absorptive gold nanorod core and magnetic nanospheres, which can potentially accumulate and concentrate targeted cells while simultaneously enhancing their specific contrast compared to background signals. In this study, HeLa cells molecularly targeted using nanocomposites with folic acid mimicking targeted rare circulating tumor cells (CTCs) were circulated at a 6 ml/min flow rate for trapping and imaging studies. Preliminary results show that the cells accumulate rapidly in the presence of an externally applied magnetic field produced by a dual magnet system. The sensitivity of the current system can reach up to 1 cell/ml in clear water. By manipulating the trapped cells magnetically, the specificity of detecting cells in highly absorptive ink solution can be enhanced with 16.98 dB background suppression by applying motion filtering on PA signals to remove unwanted background signals insensitive to the magnetic field. The results appear promising for future preclinical studies on a small animal model and ultimate clinical detection of rare CTCs in the vasculature.

A new technique using pulsed laser heating of a nanocomposite contrast agent resulting in local bubble formation and concomitant harmonic generation in a scattered probe ultrasound (US) beam is proposed to increase specific contrast in both US imaging and laser-induced photoacoustic (PA) imaging. The composite combines an emulsion bead core with amphiphilic gold nanospheres (GNSs) assembled at the interface. Clustered GNSs result in a broadened absorption spectrum in the near infrared range (700-1000 nm) compared to the typical 520 nm peak of distributed GNSs, enabling their use at depth in tissue. Illuminating the composite with a pulsed laser with appropriately chosen parameters heats the composite through optical absorption by the GNSs and results in a phase transition of the emulsion bead to form a transient bubble. By delivering a probe US pulse simultaneously, or immediately after the laser pulse is delivered, harmonic signals are produced in the scattered US beam. The results show that a residual signal created by subtracting a US signal from the simultaneous US/laser probing signal of the emulsion bead sample is 1.7 dB higher than the laser alone generated PA signal and 20 dB higher than the PA signal of a control homogeneous GNSs dispersion with the same optical absorption, indicating the nonlinear contrast enhancement from bubble dynamics. The proposed technique of local activation of this designed contrast agent can be used to dramatically enhance both the specificity and sensitivity of integrated US/PA molecular imaging.

Photoacoustic (PA) imaging has been demonstrated to be a promising modality in molecular imaging for detection of nanoparticle-targeted diseased cells or tissues. However, intrinsic absorbers, such as blood, produce strong PA background signals that severely degrade the detection sensitivity and specificity of targeted objects. Magnetomotive photoacoustic (mmPA) imaging, a newly developed molecular imaging modality, introduced dynamic manipulation into traditional PA imaging. Unlike conventional PA imaging, magnetomotive manipulation with simultaneous ultrasound/PA imaging of agents incorporating magnetic nanoparticles enables direct visualization of the signal generating object and can dramatically reduce background signals from strong optical absorbers. This paper briefly reviews recent developments in mmPA imaging, including uses of composite contrast agent, design of magnet system, and data processing for motion filtering. The use of mmPA imaging in detecting rare circulating tumor cells in blood vessels, which remains a big challenge for real-time in vivo examination using current methodologies, was also addressed. Fusion Images (ultrasound + photoacousitc) of manipulation of magnetic-gold nanorod contrast agent targeted polystyrene beads in ink solution flowing with a rate of 12 ml/min [1]. Images (a) and (b) were acquired with a cone magnet array at two different positions, and (c) is the differential image of (a) and (b). A bipolar color scale is used to display the sign of the difference. Triangles indicate the magnet positions.

Contrast-enhanced photoacoustic (PA) imaging has been proposed to identify circulating metastatic cancer cells magnetically trapped in the vasculature. However, its sensitivity is limited by the presence of a strong blood-background signal. This technique can be further improved by the significant suppression of blood background. In the phantom study presented here, significant background suppression is demonstrated with magnetomotive photoacoustic imaging. Magnetic particles with a mean diameter of 10 μm were integrated (concentration of 0.05 mg / ml) into an ink-water solution with an optical absorption coefficient of 5 cm − 1 to mimic cells targeted with magnetic nanoparticles and magnetically trapped in the human vasculature. Two mechanically moveable permanent magnets were used to accumulate microparticles in the investigated solution and manipulate them within a thin, 1.6-mm-diameter Teflon tube mimicking a blood vessel. Our results clearly indicate that the undesirable background can be effectively suppressed using the difference of PA images corresponding to different locations of accumulated particles.

Trapping and manipulation of micro-scale objects mimicking metastatic cancer cells in a flow field have been demonstrated with magnetomotive photoacoustic (mmPA) imaging. Coupled contrast agents combining gold nanorods (15 nm × 50 nm; absorption peak around 730 nm) with 15 nm diameter magnetic nanospheres were targeted to 10 μm polystyrene beads recirculating in a 1.6 mm diameter tube mimicking a human peripheral vessel. Targeted objects were then trapped by an external magnetic field produced by a dual magnet system consisting of two disc magnets separated by 6 cm to form a polarizing field (0.04 Tesla in the tube region) to magnetize the magnetic contrast agents, and a custom designed cone magnet array with a high magnetic field gradient (about 0.044 Tesla/mm in the tube region) producing a strong trapping force to magnetized contrast agents. Results show that polystyrene beads linked to nanocomposites can be trapped at flow rates up to 12 ml/min. It is shown that unwanted background in a photoacoustic image can be significantly suppressed by changing the position of the cone magnet array with respect to the tube, thus creating coherent movement of the trapped objects. This study makes mmPA imaging very promising for differential visualization of metastatic cells trafficking in the vasculature.

Magnetic trapping and manipulation of polystyrene beads mimicking magnetically targeted circulating tumor cells (CTCs) has been demonstrated using magnetomotive photoacoustic (mmPA) imaging. These techniques appear promising for the detection of rare CTCs in the vasculature.

Current systems designed for deep photoacoustic (PA) imaging typically use a low repetition rate, high power pulsed laser to provide a ns-scale pulse illuminating a large tissue volume. Acoustic signals recorded on each laser firing can be used to reconstruct a complete 2-D (3-D) image of sources of heat release within that region. Using broad-beam excitation, the maximum frame rate of the imaging system is restricted by the pulse repetition rate of the laser. An alternate illumination approach is proposed based on fast scanning by a low energy (~ 1 mJ) high repetition rate (up to a few kHz) narrow laser beam (~1 mm) along the tissue surface over a region of interest. A final PA image is produced from the summation of individual PA images reconstructed at each laser beam position. This concept can take advantage of high repetition rate fiber lasers to create PA images with much higher frame rates than current systems, enabling true real-time integration of photoacoustics with ultrasound imaging. As an initial proof of concept, we compare conventional broad beam illumination to a scanned beam approach in a simple model system. Two transparent teflon tubes with diameters of 1.6 mm and 0.8 mm were filled with ink having an absorption coefficient of 5 cm -1 . These tubes were buried inside chicken breast tissue acting as an optical scattering medium. They were separated by 3 mm or 10 mm to test spatial and contrast resolution for the two scan formats. The excitation wavelength was 700 nm. The excitation source is a traditional OPO pumped by a Q-switched Nd:YAG laser with doubler. Photoacoustic images were reconstructed using signals from a small, scanned PVDF transducer acting as an acoustic array. Two different illumination schemes were compared: one was 15 mm x 10 mm in cross section and acted as the broad beam; the other was 5 mm x 2 mm in cross section (15 times smaller than the broad beam case) and was scanned over an area equivalent to broad beam illumination. Multiple images obtained during narrow beam scanning were added together to form one PA image equivalent to the single-shot broad beam one. Results of the phantom study indicate that PA images formed by narrow beam scanning excitation can be equivalent to one shot broad beam illumination in signal to noise ratio and spatial resolution. Future studies will focus on high repetition-rate laser sources and scan formats appropriate for real-time, integrated deep photoacoustic/ultrasonic imaging.

Photoacoustic tomography (PAT) has garnered much attention for its high contrast and excellent spatial resolution of perfused tissues. Gold nanorods (GNRs) have been employed to further enhance the imaging contrast of PAT. However, the photon fluences typically needed for PA wave induction often also result in GNR shape changes that significantly reduce the efficiency of acoustic wave generation. In this work, we propose, synthesize, and evaluate amorphous silica-coated gold nanorods (GNR-Si) in an effort to improve contrast agent stability and ameliorate efficiency loss during photoacoustic (PA) wave induction. TEM and optical absorption spectra measurements of GNR and GNR-Si show that encasing GNRs within amorphous silica provides substantial protection of nanorod conformation from thermal deformation. PA signals generated by GNR-Si demonstrate considerably greater resistance to degradation of signal intensity with repetitive pulsing than do uncoated GNRs, thereby enabling much longer, high-contrast imaging sessions than previously possible. The prolongation of high-contrast imaging, and biocompatibility and easy surface functionalization for targeting ligands afforded by amorphous silica, suggest GNR-Si to be potentially significant for the clinical translation of PAT.

Gold nanoparticles have been used as contrast agent in photoacoustic imaging to increase the detection sensitivity. For example, gold nanorods (AuNRs) have been used in time-intensity based flow estimation and used as nanoprobes to target cancer cells for early diagnosis and effective treatment. In this study, we aimed at the design and synthesis of a new type of gold nanoparticles with enhanced photoacoustic response. The key hypothesis is to create a nanostructure that allows anisotropic heat release. Specifically, such a structure results in higher heat flux transmitting outwards from the ends of the particle and therefore a greater temperature gradient can be created. To achieve this, a layer of SiO2 was coated along the longer axis of the gold nanorods, leaving both ends uncovered. These new particles are labeled as AuNR@nu-SiO2 with non-uniform ("nu") coating of silica. Experiments were performed to demonstrate the enhanced photoacoustic response from AuNR@nu-SiO2. The optical illumination was delivered by a Ti: Sapphire laser pumped by an Nd:YAG laser. A home-made photoacoustic transducer with a center frequency of 20 MHz detected the resulted acoustic signal. First, new types of particles coated with and without SiO2 were compared to bare AuNR in order to show the ability of the new nanostructure to enhance photoacoustic response. Second, the shape stability of the new particles was evaluated by measuring the photoacoustic responses versus time after high power laser irradiation. Third, the effect of thickness of SiO2 of AuNR@nu-SiO2 ranges from 1 nm to 15 nm was also evaluated. Results show that the mean intensity in photoacoustic image increase about 5 dB for AuNR@nu-SiO2 compared to bare AuNR. Also, it reveals that the normalized intensity for AuNR drops to below 0.6 while it is mostly larger than 0.7 in the case of AuNR@nu-SiO2 under pulse laser irradiation. In other words, the new type of nanoparticles is less susceptible to shape transformation. Moreover, it is indicated that the photoaocustic response increases slightly with the thickness of SiO2 and approach to an maximum response at 9 nm thickness. In short, these new particles can be used to achieve the same level of photoacoustic response with a fewer amount of particles, which means that there is less toxicity.

Plasmonic photothermal therapy is a new cancer thermotherapy method based on surface plasmon resonance of nanoparticles. It is important to measure the temperature during thermotherapy for safety and efficacy. In this study, we apply a photoacoustic (PA) method for real-time, non-invasive temperature measurements. In particular, this method can be effectively combined with a photothermal therapy system that we developed in parallel. The method is based on the fact that the PA pressure amplitude is linearly related to temperature. To explore its potential, a home-made, 20 MHz PA transducer was used, in which an optical fiber was inserted in its center for emitting laser pulses while the PA signal was simultaneously detected. Continuous wave (CW) laser was used to heat the subject, including both phantoms and mice. The temperature of the region of interest was also measured by a fine-needle thermal couple. Results show that the temperature was linearly proportional to the PA signal with good correlation with the CW laser irradiation. The in vivo study also demonstrated potential of this technique.

Contrast in photoacoustic imaging is primarily determined by optical absorption. This paper proposes a subband imaging method to further enhance the image contrast. The method is based on media with different absorptions generating acoustic waves with different frequency contents. Generally, assuming all other conditions remain the same, a high-absorption medium generates acoustic waves with higher frequency components, and hence the imaging contrast can be enhanced by appropriate selection of the spectral subbands. This study employed both finite-difference, time-domain-based simulations and phantom imaging. The numerical results show that the peak frequencies of the signals for objects with absorption coefficients of 1 and 100 cmM(-1) were 2.4 and 7.8 MHz, respectively. Imaging an agar-based phantom further demonstrated that the contrast between two objects with absorption coefficients of 5.01 and 41.75 cm(-1) can be improved by 4-10 dB when the frequency band was changed from 0-7 to 7-14 MHz. Finally, a method to further enhance the contrast based on optimal weighting is also presented. The proposed method is of particular interest in photoacoustic molecular imaging.

A quantitative flow measurement method that utilizes a sequence of photoacoustic images is described. The method is based on the use of gold nanorods as a contrast agent for photoacoustic imaging. The peak optical absorption wavelength of a gold nanorod depends on its aspect ratio, which can be altered by laser irradiation (we establish a wash-in flow estimation method of this process). The concentration of nanorods with a particular aspect ratio inside a region of interest is affected by both laser-induced shape changes and replenishment of nanorods at a rate determined by the flow velocity. In this study, the concentration is monitored using a custom-designed, high-frame-rate photoacoustic imaging system. This imaging system consists of fiber bundles for wide area laser irradiation, a laser ultrasonic transducer array, and an ultrasound front-end subsystem that allows acoustic data to be acquired simultaneously from 64 transducer elements. Currently, the frame rate of this system is limited by the pulse-repetition frequency of the laser (i.e., 15 Hz). With this system, experimental results from a chicken breast tissue show that flow velocities from 0.125 to 2 mms can be measured with an average error of 31.3%.

In this study, photoacoustic flow measurement methods based on wash-in analysis are presented. These methods use the rod-to-sphere shape transformations of gold nanorods induced by pulsed-laser irradiation. Due to the shape dependence of the optical absorption of the gold nanorods, these shape transitions are associated with a change in the peak optical absorption wavelength. Pulsed-laser irradiation at the wavelength corresponding to the peak optical absorption of the original gold nanorods allows the particles that undergo shape changes to be viewed as "being destructed" by the laser irradiation at that wavelength, hence, flow information can be derived from the change in ultrasound intensity that is directly related to the wash-in rate of the gold nanorods and the laser intensity. Two flow estimation methods based on the wash-in analysis are described. The first method first applies high-energy laser pulses that induce shape changes in all the nanorods. A series of low-energy pulses then are applied to monitor the acoustic signal change as new nanorods flow into the region of interest. The second method uses single-energy laser pulses such that the "destruction" and "detection" are performed simultaneously. The simulation results show that it is valid to fit the time-intensity curves by exponential models. To demonstrate the validity of the proposed methods, an Nd:YAG pulsed laser operating at 1064 nm was used for optical irradiation, and a 1-MHz ultrasonic transducer was used for acoustic detection. Gold nanorods with a peak optical absorption at 1018 nm and a concentration of 0.26 nM were used to estimate flow velocities ranging from 0.35 to 2.83 mm/s. The linear regression results show that the correlation coefficients between the measured velocities and the true values are close to unity (or = 0.94), thus demonstrating the feasibility of the proposed photoacoustic techniques for relative flow estimation.

In this paper, the off-state breakdown characteristics of two different AlGaN/GaN high electron mobility transistors (HEMTs), featuring a 50-nm and a 150-nm GaN thick channel layer, respectively, are compared. The HEMT with a thick channel exhibits a little larger pinch-off drain current but significantly enhanced off-state breakdown voltage (BVoff). Device simulation indicates that thickening the channel increases the drain-induced barrier lowering (DIBL) but reduces the lateral electric field in the channel and buffer underneath the gate. The increase of BVoff in the thick channel device is due to the reduction of the electric field. These results demonstrate that it is necessary to select an appropriate channel thickness to balance DIBL and BVoff in AlGaN/GaN HEMTs.

The hypothesis that quantitative blood flow measurements are feasible with the time-intensity based method in photoacoustic imaging using gold nanoparticles as contrast agent is experimentally tested. The in vitro results show good linearity between the measurements and the theory, thus suggesting the potential of relative photoacoustic flow measurements with gold nanoparticles.

We have previously tested the hypothesis that gold nanospheres can be used for quantitative flow estimation in optoacoustic imaging based on wash-out analysis. In this study, a wash-in flow analysis method using gold nanorods is developed. Specifically, the proposed technique makes use of the shape dependence of the optical absorption wavelength of gold nanorods and their shape transformation after pulsed laser irradiation. With pulsed laser irradiation at a wavelength that corresponds to the peak optical absorption of the original shape of the gold nanorods, the particles that undergo shape changes can be viewed as "being destructed" by laser irradiation at that wavelength. Based on this property, a destruction-mode technique was developed. With this technique, a destruction laser pulse is followed by a series of observation pulses. The linear regression results show that the correlation coefficient between the measured velocities and the true values was close to unity (0.96), thus demonstrating the feasibility of the proposed destruction-mode technique for quantitative flow estimation in optoacoustic imaging. .

Cancer cells presented altered surface molecules to encourage their growth and metastasis. Expression of oncogeneic surface molecules also play important roles in the prediction of clinical outcome and treatment response of anti-cancer drugs. It is thus conceivable that imaging of cancer lesions while simultaneously obtaining their pathogenic information at molecular level of as many oncogenic proteins as possible is of great clinical significance. Gold nanoparticles have been used as a contrast agent for photoacoustic imaging. In addition, gold nanoparticles can be bioconjugated to probe certain molecular processes. An intriguing property of gold nanoparticles is its ability to tailor its optical properties. For example, size effects on the surface plasmon absorption of spherical gold nanoparticles have shown that the peak optical absorption red-shifts with the increasing particle size. In addition, the optical absorption spectrum of cylindrical gold nanoparticles (i.e., gold nanorods) exhibits a strong absorption band that is directly related to the aspect ratio. With these unique characteristics, selective targeting can be achieved in photoacoustic molecular imaging. Specifically, gold nanorods with different aspect ratios can be bioconjugated to different antibodies. Multiple targeting and simultaneous detection can then be achieved by using laser irradiation at the respective peak optical absorption wavelength. In this study, photoacoustic multiple targeting using gold nanorods is experimentally demonstrated. We have chosen Her2 and CXCR4 as our primary target molecule as Her2 expression is associated with growth characteristics and sensitivity to Herceptin chemotherapy. On the other hand, CXCR4 expression predict the organ-specific metastatic potential of the cancer cells for clinical intervention in advance. Monoclonal antibody (mAb) against Her2/neu was conjugated to nanorods with several different aspect ratios. The agarose gel is suitable for photoacoustic signal acquisition. A wavelength tunable Ti-Sapphire laser was used for laser irradiation and a 1 MHz ultrasound transducer was used for acoustic detection. The optical wavelength of the laser was tuned between 800 nm and 940 nm, corresponding to gold nanorods of an aspect ratio ranging from 3.7 to 5.9. The results clearly show the potential of photoacoustic molecular imaging with multiple targeting in revealing different oncogene expression levels of the cancer cells.

A high frame rate photoacoustic imaging system is described. Applications of this system to perfusion measurements are also presented as a demonstration of its potential usage. The system consists of an ultrasound front-end sub-system for acquisition of acoustic array data. The ultrasound front-end sub-system is also known as the DiPhAS (digital phased array system) which is capable of simultaneously acquiring radio frequency data from 64 transducer channels at a rate up to 40 MSamples/sec per channel. In this study, an ultrasonic linear array with a 5 MHz center frequency was employed as part of the integrated photoacoustic probe. The photoacoustic probe also had two linear light guides mounted on the sides of the ultrasonic array for broad laser irradiation from a Q-switched Nd:YAG pulsed laser. After the acquired ultrasound array data were transferred to a personal computer via a high speed digital I/Q card, dynamic focusing and image reconstruction were done off-line. The 64-channel array data can be acquired and transferred every 4 milliseconds, thus making the frame rate of the system up to 250 Hz. The actual frame rate of the current system is limited by the pulse repetition frequency of the laser at 15 Hz. To demonstrate capabilities of the system, photoacoustic perfusion measurements with gold nanorods were performed. A previously proposed time-intensity based flow estimation technique utilizing the shape transitions of gold nanorods under laser irradiation was employed. Good estimation results were achieved and potential of this high frame rate photoacoustic imaging system is clearly demonstrated.

A time-intensity based method for photoacoustic blood flow measurements was proposed in last year's meeting. The method made use of the strong photoacoustic response of gold nanospheres and the "wash-out" characteristics of the nanospheres were analyzed. In this paper, we develop a new quantitative technique for measuring blood flows based on the "wash-in" characteristics of the nanoparticles. In particular, the technique makes use of the shape dependence of the optical absorption of gold nanorods (i.e., cylindrical nanoparticles) and the transitions in their shape induced by pulsed laser irradiation. The photon-induced shape transition of gold nanorods involves mainly a rod-to-sphere conversion and a shift in the peak optical absorption wavelength. The application of a series of laser pulses with the same laser energy will induce shape changes in gold nanorods as they flow through a region of interest, with quantitative flow information being derived from the photoacoustic signals from the irradiated gold nanorods measured as a function of time. To demonstrate the feasibility of the technique, an Nd:YAG laser operating at 1064 nm was used for irradiation and a ultrasonic transducer with a center frequency of 1 MHz was used for acoustic detection. Excellent agreement between the measured velocities and the actual velocities was demonstrated, with a linear regression correlation coefficient higher than 0.9. Compared to the wash-out analysis, the wash-in analysis is more suitable for measuring flows in microcirculation.

A quantitative technique for flow measurements based on a wash-in analysis is proposed. The technique makes use of the shape dependence of the optical absorption of gold nanorods and the transitions in their shape induced by pulsed laser irradiation. The photon-induced shape transition of gold nanorods involves mainly a rod-to-sphere conversion and a shift in the peak optical absorption wavelength. The application of a series of laser pulses will induce shape changes in gold nanorods as they flow through a region of interest, with quantitative flow information being derived from the photoacoustic signals from the irradiated gold nanorods measured as a function of time. To demonstrate the feasibility of the technique, a Nd:YAG laser operating at 1064 nm was used for irradiation and a 1 MHz ultrasonic transducer was used for acoustic detection. The flow velocity ranged from 0.35 to 2.83 mm/s. Excellent agreement between the measured velocities and the actual velocities was demonstrated, with a linear regression correlation coefficient of 0.93. This study is a pioneer work on wash-in flow estimation in photoacoustic imaging.

The indicator-dilution theory has been used for flow rate measurements in various imaging modalities, including magnetic resonance imaging, computed tomography and ultrasound. The experimental procedure generally involves the injection of a dose of indicator (i.e., the contrast agent), after which the concentration of the agent is monitored as a function of time; it is therefore also known as the time-intensity method. Although the time-intensity method has been widely applied to other imaging modalities, it has not been demonstrated with optoacoustic imaging. In this study, we experimentally test the hypothesis that quantitative blood flow measurements are feasible with the time-intensity based method in optoacoustic imaging. Gold nanospheres (broad band absorption spectrum peaks at 520 nm) were used as the optoacoustic contrast agent. The imaging system consisted of a frequency-doubled Nd:YAG laser operating at 532 nm for optical illumination, and an ultrasonic single crystal transducer with a center frequency of 3.5 MHz and a focal depth of 7 cm for detection. The volumetric flow rate ranged from 0.23 to 4.29 ml/sec, and the volume of the mixing chamber was from 30 to 80 ml. Results show good agreement between the measured mean transit times and the predicted time constants (correlation coefficient higher than 0.88), thus demonstrating the feasibility of the time-intensity based flow measurement technique. In addition to describing the method and experimental results, issues regarding the system sensitivity and estimation of the dilution transfer function are also discussed.

Laser ultrasonic (LU) inspection represents an attractive, non-contact method to evaluate composite materials. Current non-contact systems, however, have relatively low sensitivity compared to contact piezoelectric detection. They are also difficult to adjust, very expensive, and strongly influenced by environmental noise. Here, we demonstrate that most of these drawbacks can be eliminated by combining a new generation of compact, inexpensive fiber lasers with new developments in fiber telecommunication optics and an optimally designed balanced probe scheme. In particular, a new type of a balanced fiber-optic Sagnac interferometer is presented as part of an all-optical LU pump-probe system for non-destructive testing and evaluation of aircraft composites. The performance of the LU system is demonstrated on a composite sample with known defects. Wide-band ultrasound probe signals are generated directly at the sample surface with a pulsed fiber laser delivering nanosecond laser pulses at a repetition rate up to 76 kHz rate with a pulse energy of 0.6 mJ. A balanced fiber-optic Sagnac interferometer is employed to detect pressure signals at the same point on the composite surface. A- and B-scans obtained with the Sagnac interferometer are compared to those made with a contact wide-band polyvinylidene fluoride transducer.

Step-stress experiments are performed in this paper to investigate the degradation mechanism of an AlGaN/GaN high electron mobility transistor (HEMT). It is found that the stress current shows a recoverable decrease during each voltage step and there is a critical voltage beyond which the stress current starts to increase sharply in our experiments. We postulate that defects may be randomly induced within the AlGaN barrier by the high electric field during each voltage step. But once the critical voltage is reached, the trap concentration will increase sharply due to the inverse piezoelectric effect. A leakage path may be introduced by excessive defect, and this may result in the permanent degradation of the AlGaN/GaN HEMT.

In recent years, conjugated polymers have attracted considerable attention from the imaging community as a new class of contrast agent due to their intriguing structural, chemical, and optical properties. Their size and emission wavelength tunability, brightness, photostability, and low toxicity have been demonstrated in a wide range of in vitro sensing and cellular imaging applications, and have just begun to show impact in in vivo settings. In this Perspective, we summarize recent advances in engineering conjugated polymers as imaging contrast agents, their emerging applications in molecular imaging (referred to as in vivo uses in this paper), as well as our perspectives on future research.

Results on magnetically trapping and manipulating micro-scale beads circulating in a flow field mimicking metastatic cancer cells in human peripheral vessels are presented. Composite contrast agents combining magneto-sensitive nanospheres and highly optical absorptive gold nanorods were conjugated to micro-scale polystyrene beads. To efficiently trap the targeted objects in a fast stream, a dual magnet system consisting of two flat magnets to magnetize (polarize) the contrast agent and an array of cone magnets producing a sharp gradient field to trap the magnetized contrast agent was designed and constructed. A water-ink solution with an optical absorption coefficient of 10 cm⁻¹ was used to mimic the optical absorption of blood. Magnetomotive photoacoustic imaging helped visualize bead trapping, dynamic manipulation of trapped beads in a flow field, and the subtraction of stationary background signals insensitive to the magnetic field. The results show that trafficking micro-scale objects can be effectively trapped in a stream with a flow rate up to 12 ml/min and the background can be significantly (greater than 15 dB) suppressed. It makes the proposed method very promising for sensitive detection of rare circulating tumor cells within high flow vessels with a highly absorptive optical background.

Kink effect is analyzed in AlGaN/GaN devices primarily. A semiempirical model is given by analyzing the kink effect on AlGaN/GaN high electron mobility transistor and by considering the relationship between Vds,kink and gate voltage. Due to a little error between simulation results and measured data, this model can be used to identify the occurrence of kink effect and change in drain current. The analyses of experimental results and model simulation lead to a conclusion that impact ionization plays an important role in generating kink effect.

Several imaging modalities such as optical coherence tomography, photothermal, photoacoustic and magnetic resonance imaging, are sensitive to different physical properties (i.e. scattering, absorption and magnetic) that can provide contrast within biological tissues. Usually exogenous agents are designed with specific properties to provide contrast for these imaging methods. In nano-biotechnology there is a need to combine several of these properties into a single contrast agent. This multifunctional contrast agent can then be used by various imaging techniques simultaneously or can be used to develop new imaging modalities. We reported and characterized a multifunctional nanoparticle, made from gold nanoshells, which exhibits scattering, photothermal, photoacoustic, and magnetic properties.

The use of gold nanorods for photoacoustic molecular imaging in vivo with simultaneous multiple selective targeting is reported. The extravasation of multiple molecular probes is demonstrated, and used to probe molecular information of cancer cells. This technique allows molecular profiles representing tumor characteristics to be obtained and a heterogeneous population of cancer cells in a lesion to be determined. The results also show that the image contrast can be enhanced by using a mixture of different molecular probes. In this study, HER2, EGFR, and CXCR4 were chosen as the primary target molecules for examining two types of cancer cells, OECM1 and Cal27. OECM1 cells overexpressed HER2 but exhibited a low expression of EGFR, while Cal27 cells showed the opposite expression profile. Single and double targeting resulted in signal enhancements of up to 3 dB and up to 5 dB, respectively, and hence has potential in improving cancer diagnoses.

Integrating high contrast bubbles from ultrasound imaging with plasmonic absorbers from photoacoustic imaging is investigated. Nanoemulsion beads coated with gold nanopsheres (NEB-GNS) are excited with simultaneous light (transient heat at the GNS's) and ultrasound (rarefactional pressure) resulting in a phase transition achievable under different scenarios, enhancing laser-induced acoustic signals and enabling specific detection of nanoprobes at lower concentration. An automated platform allowed dual parameter scans of both pressure and laser fluence while recording broadband acoustic signals. Two types of NEB-GNS and individual GNS were investigated and showed the great potential of this technique to enhance photoacoustic/acoustic signals. The NEB-GNS size distribution influences vaporization thresholds which can be reached at both permissible ultrasound and light exposures at deep penetration and at low concentrations of targets. This technique, called sono-photoacoustics, has great potential for targeted molecular imaging and therapy using compact nanoprobes with potentially high-penetrability into tissue.